重组Anti-Histone H3抗体[EPR16987] -核Marker and ChIP Grade

• ChIP

Unknown

ChIP - Anti-Histone H3 antibody [EPR16987] - Nuclear Marker and ChIP Grade (AB176842)

Chromatin was prepared from HeLa cells according to the Abcam X-ChIP protocol. Cells were fixed with formaldehyde for 10 minutes. The ChIP was performed with 25μg of chromatin, 2μg of ab176842 (red), and 20μl of Protein A/G sepharose beads. No antibody was added to the beads control (grey). The immunoprecipitated DNA was quantified by real time PCR (Taqman approach for active and inactive loci, Sybr green approach for heterochromatic loci). Primers and probes are located in the first kb of the transcribed region.

• WB

Supplier Data

Western blot - Anti-Histone H3 antibody [EPR16987] - Nuclear Marker and ChIP Grade (AB176842)

Blocking and diluting buffer and concentration : 5% NFDM/TBST.

Performed under reducing conditions.

In Western blot, ab324563 was shown to bind specifically to NEDD4. Target of interest was observed at 115 kDa in wild-type HAP1 cell lysates (lane 2) with no signal observed at this size in NEDD4 knockout cell line (lane 1) (lane 1, knockout cell line ab

ab256783).

To minimize protein degradation, cells/tissues were lysed immediately after harvest and then applied to a gel and transfer membrane for Western blotting as soon as possible.

The samples are unboiled.

All lanes:

Western blot - Anti-NEDD4 antibody [EPR30127-570] (<a href='/products/primary-antibodies/nedd4-antibody-epr30127-570-ab324563'>ab324563</a>) at 1/1000 dilution

Lane 1:

NEDD4 knockout parental HAP1 (human chronic myelogenous leukemia near-haploid cell ) whole cell lysate at 20 µg

Lane 2:

Parental HAP1 whole cell lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/100000 dilution

Observed band size: 115 kDa,15 kDa

false

Exposure time: 37s

• WB

Supplier Data

Western blot - Anti-Histone H3 antibody [EPR16987] - Nuclear Marker and ChIP Grade (AB176842)

The expression of TOPBP1 S1138 is upregulated in response to hydroxyurea treatment (PMID : 17446169).

Lysates were freshly made and used for Western blotting immediately to minimize protein degradation.

In Western blot, Anti-Histone H3 antibody [EPR16987] - Nuclear Marker and ChIP Grade (ab176842) staining at 1/100000 dilution.

In Western blot, Anti-TOPBP1 antibody staining at 1/1000 dilution.

All lanes:

Western blot - Anti-TopBP1 (phospho S1138) antibody [EPR27056-2] (<a href='/products/primary-antibodies/topbp1-phospho-s1138-antibody-epr27056-2-ab316746'>ab316746</a>) at 1/1000 dilution

Lane 1:

Untreated U-2 OS (human bone osteosarcoma epithelial cell) whole cell lysate (untreated membrane) at 30 µg with 5% NFDM/TBST

Lane 2:

U-2 OS treated with 2mM hydroxyurea for 24h whole cell lysate (untreated membrane) at 30 µg with 5% NFDM/TBST

Lane 3:

U-2 OS treated with 2mM hydroxyurea for 24h whole cell lysate (alkaline phosphatase treated membrane) at 30 µg with 5% NFDM/TBST

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/20000 dilution

Observed band size: 170 kDa,15 kDa

false

Exposure time: 92s

• WB

Supplier Data

Western blot - Anti-Histone H3 antibody [EPR16987] - Nuclear Marker and ChIP Grade (AB176842)

Blocking and dilution buffer : 5% NFDM/TBST

All lanes:

Western blot - Anti-Histone H3 antibody [EPR16987] - Nuclear Marker and ChIP Grade (ab176842) at 1/10000 dilution

All lanes:

HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/50000 dilution

Predicted band size: 15 kDa

Observed band size: 15 kDa

false

Exposure time: 3s

• WB

Lab

Western blot - Anti-Histone H3 antibody [EPR16987] - Nuclear Marker and ChIP Grade (AB176842)

Blocking and diluting buffer and concentration : 5% NFDM/TBST. ab181602 was used as GAPDH loading control. ab176842 was for Histone H3 detection.

All lanes:

Western blot - Anti-Histone H3 (acetyl K9) antibody [Y28] - ChIP Grade (<a href='/products/primary-antibodies/histone-h3-acetyl-k9-antibody-y28-chip-grade-ab32129'>ab32129</a>) at 1/10000 dilution

Lane 1:

HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysate at 15 µg

Lane 2:

HeLa (Human cervix adenocarcinoma epithelial cell) treated with 500ng/ml Trichostatin A for 4 hours whole cell lysate at 15 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/20000 dilution

Predicted band size: 15 kDa

Observed band size: 15 kDa

false

• WB

Supplier Data

Western blot - Anti-Histone H3 antibody [EPR16987] - Nuclear Marker and ChIP Grade (AB176842)

The molecular weight observed is consistent with what has been described in the literature (PMID : 38240850).

The identity of the lower MW bands are unknown.

In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) staining at 1/200000 dilution.

In Western blot, Anti-Histone H3 antibody [EPR16987] - Nuclear Marker and ChIP Grade (ab176842) staining at 1/100000 dilution.

All lanes:

Western blot - Anti-SOX4 antibody [EPR28132-51] (<a href='/products/primary-antibodies/sox4-antibody-epr28132-51-ab316850'>ab316850</a>) at 1/1000 dilution

Lane 1:

DLD-1 (human colorectal adenocarcinoma epithelial cell) nuclear fraction at 20 µg with 5% NFDM/TBST

Lane 2:

DLD-1 cytoplasmic fraction at 20 µg with 5% NFDM/TBST

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/20000 dilution

Observed band size: 70 kDa,36 kDa,15 kDa

false

Exposure time: 59s

• WB

Supplier Data

Western blot - Anti-Histone H3 antibody [EPR16987] - Nuclear Marker and ChIP Grade (AB176842)

Blocking and diluting buffer and concentration : 5% NFDM/TBST.

In Western blot, Anti-Histone H3 antibody [EPR16987] - Nuclear Marker and ChIP Grade (ab176842) staining at 1/100000 dilution.

In Western blot, Anti-TRAF1 antibody [EPR26204-112] (ab300075) staining at 1/1000 dilution.

All lanes:

Western blot - Anti-TRAF1 (phospho S146) antibody [EPR25987-11] (<a href='/products/primary-antibodies/traf1-phospho-s146-antibody-epr25987-11-ab315154'>ab315154</a>) at 1/1000 dilution

Lane 1:

Untreated Raji (human Burkitts lymphoma B lymphocyte) whole cell lysate (untreated membrane) at 20 µg

Lane 2:

Raji treated with 100 nM Calycin A for 25 minutes whole cell lysate (untreated membrane) at 20 µg

Lane 3:

Untreated Raji (human Burkitts lymphoma B lymphocyte) whole cell lysate (alkaline phosphatase treated membrane) at 20 µg

Lane 4:

Raji treated with 100 nM Calycin A for 25 minutes whole cell lysate (alkaline phosphatase treated membrane) at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/100000 dilution

Observed band size: 50 kDa,15 kDa

false

Exposure time: 180s

• WB

Supplier Data

Western blot - Anti-Histone H3 antibody [EPR16987] - Nuclear Marker and ChIP Grade (AB176842)

Blocking and diluting buffer and concentration : 5% NFDM/TBST

In Western blot, anti- H3 antibody (ab176842) loading control staining at 1/100000 dilution.

All lanes:

Western blot - Anti-FKBP12 antibody [EPR27367-59] (<a href='/products/primary-antibodies/fkbp12-antibody-epr27367-59-ab313378'>ab313378</a>) at 1/1000 dilution

Lane 1:

SH-SY5Y (human neuroblastoma cell line from bone marrow) cytoplasmic fraction at 20 µg

Lane 2:

SH-SY5Y nuclear fraction at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/100000 dilution

Predicted band size: 11 kDa

Observed band size: 14 kDa

false

Exposure time: 81s

• WB

Supplier Data

Western blot - Anti-Histone H3 antibody [EPR16987] - Nuclear Marker and ChIP Grade (AB176842)

Blocking and diluting buffer and concentration : 5% NFDM/TBST.

Negative control : Jurkat (PMID : 8691135, 11841448), HEK-293.

The expression profile/ molecular weight observed is consistent with what has been described in the literature (PMID : 8691135).

In lanes 1-5, the lysates were stored at -80℃ prior to Western Blotting. The bands beneath the target band (46 kDa) are likely to be degradation products. In lanes 6-7, the lysates were freshly made and used for Western Blotting immediately to minimize protein degradation.

This blot was developed using a high sensitivity ECL substrate. The high-sensitivity ECL substrate used allows for the detection of proteins in the mid-femtogram range.

In Western blot, anti- H3 antibody (ab176842) loading control staining at 1/100000 dilution.

Exposure time :

Lanes 1-5 : 48 seconds

Lanes 6-7 : 26 seconds

All lanes:

Western blot - Anti-SPIB antibody [EPR26784-117] (<a href='/products/primary-antibodies/spib-antibody-epr26784-117-ab309346'>ab309346</a>) at 1/1000 dilution

Lane 1:

Daudi (human Burkitts lymphoma lymphoblast) whole cell lysate at 20 µg

Lane 2:

HDLM-2 (human Hodgkin lymphoma) whole cell lysate at 20 µg

Lane 3:

Raji (human Burkitts lymphoma B lymphocyte) whole cell lysate at 20 µg

Lane 4:

Jurkat (human T cell leukemia T lymphocyte) whole cell lysate at 20 µg

Lane 5:

HEK-293 (human embryonic kidney epithelial cell) whole cell lysate at 20 µg

Lane 6:

Raji whole cell lysate at 20 µg

Lane 7:

Jurkat whole cell lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/100000 dilution

Observed band size: 46 kDa

true

• WB

Supplier Data

Western blot - Anti-Histone H3 antibody [EPR16987] - Nuclear Marker and ChIP Grade (AB176842)

The identity of the higher MW bands are unknown.

In Western blot, Anti-Histone H3 antibody [EPR16987] - Nuclear Marker and ChIP Grade (ab176842) staining at 1/100000 dilution.

In Western blot, Anti-MYL9 antibody [EPR13012 (2)] (ab191393) staining at 1/1000 dilution.

All lanes:

Western blot - Anti-MYL9 (pS20) + MYL12A (pS19) + MYL12B (pS20) antibody [EPR27203-65] (<a href='/products/primary-antibodies/myl9-ps20-myl12a-ps19-myl12b-ps20-antibody-epr27203-65-ab316750'>ab316750</a>) at 1/1000 dilution

Lane 1:

Untreated HeLa (human epithelial cell line from cervical adenocarcinoma) whole cell lysate (untreated membrane) at 45 µg with 5% NFDM/TBST

Lane 2:

HeLa treated with 100ng/ml Calyculin A for 15 minutes, whole cell lysate (untreated membrane) at 45 µg with 5% NFDM/TBST

Lane 3:

Untreated HeLa (human epithelial cell line from cervical adenocarcinoma) whole cell lysate (alkaline phosphatase treated membrane) at 45 µg with 5% NFDM/TBST

Lane 4:

HeLa treated with 100ng/ml Calyculin A for 15 minutes, whole cell lysate (alkaline phosphatase treated membrane) at 45 µg with 5% NFDM/TBST

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/20000 dilution

Observed band size: 20 kDa,15 kDa

false

Exposure time: 70s

• WB

Supplier Data

Western blot - Anti-Histone H3 antibody [EPR16987] - Nuclear Marker and ChIP Grade (AB176842)

Blocking and diluting buffer and concentration : 5% NFDM/TBST Negative control : 293T (PMID : 33042257). Exposure time : Lane 1 : 3 seconds; Lanes 2-4 : 26 seconds.

All lanes:

Western blot - Anti-P2X7 antibody [EPR26989-11] (<a href='/products/primary-antibodies/p2x7-antibody-epr26989-11-ab307718'>ab307718</a>) at 1/1000 dilution

Lane 1:

Human cerebellum tissue lysate at 20 µg

Lane 2:

U-87 MG (human glioblastoma-astrocytoma epithelial cell) whole cell lysate at 20 µg

Lane 3:

SK-MEL-28 (human malignant melanoma cell ) whole cell lysate at 20 µg

Lane 4:

293T (human embryonic kidney epithelial cell) whole cell lysate at 20 µg

Secondary

Lanes 1 - 4:

Goat Anti-Rabbit IgG (HRP) with minimal cross-reactivity with human IgG at 1/100000 dilution

Lanes 1 - 4:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/2000 dilution

Observed band size: 75 kDa

false

Exposure time: 3s

• WB

Lab

Western blot - Anti-Histone H3 antibody [EPR16987] - Nuclear Marker and ChIP Grade (AB176842)

Blocking buffer and concentration : 5% NFDM/TBST  Diluting buffer and concentration : 5% NFDM/TBST ab176323 detects a higher band in nuclear fraction than in non-nuclear fraction, which might be the correct band of interest. ab176842 and ab181602 were used as loading controls.

All lanes:

Western blot - Anti-LXR alpha antibody [EPR6508(N)] (<a href='/products/primary-antibodies/lxr-alpha-antibody-epr6508n-ab176323'>ab176323</a>) at 1/1000 dilution

Lane 1:

Jurkat (Human T cell leukemia T lymphocyte) cells non-nuclear fraction at 20 µg

Lane 2:

Jurkat (Human T cell leukemia T lymphocyte) cells nuclear fraction at 20 µg

Lane 3:

SH-SY5Y (Human neuroblastoma epithelial cell) cells non-nuclear fraction at 20 µg

Lane 4:

SH-SY5Y (Human neuroblastoma epithelial cell) cells nuclear fraction at 20 µg

Lane 5:

IMR-32 (Human neuroblastoma neuroblast) cells non-nuclear fraction at 20 µg

Lane 6:

IMR-32 (Human neuroblastoma neuroblast) cells nuclear fraction at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/20000 dilution

Predicted band size: 50 kDa

Observed band size: 50 kDa

false

Exposure time: 7s

• WB

Supplier Data

Western blot - Anti-Histone H3 antibody [EPR16987] - Nuclear Marker and ChIP Grade (AB176842)

Lane 1:

HaCaT (human skin keratinocyte) whole cell lysate

Lane 2:

HCC1937 (human mammary gland epithelial cell) whole cell lysate at 20 µg

Observed band size: 93 kDa,63 kDa

false

Exposure time: 180s

• WB

Lab

Western blot - Anti-Histone H3 antibody [EPR16987] - Nuclear Marker and ChIP Grade (AB176842)

Blocking and diluting buffer and concentration : 5% NFDM/TBST. ab181602 was used as GAPDH loading control. Lane 1 - Lane 4 : ab176842 was used for Histone H3 detection. The molecular weight observed is consistent with what has been described in the literatures (PMID : 15608675, PMID : 13679379).

All lanes:

Western blot - Anti-FRA1 antibody [EP4711] (<a href='/products/primary-antibodies/fra1-antibody-ep4711-ab124722'>ab124722</a>) at 1/1000 dilution

Lane 1:

T-47D (Human ductal breast epithelial tumor epithelial cell) nuclear fraction cell lysate at 20 µg

Lane 2:

T-47D without nuclear fraction cell lysate at 20 µg

Lane 3:

MCF7 (Human breast adenocarcinoma epithelial cell) whole cell lysate at 20 µg

Lane 4:

T-47D (Human ductal breast epithelial tumor epithelial cell) whole cell lysate at 20 µg

Lane 5:

MDA-MB-435S (Human mammary gland ductal carcinoma melanocyte) whole cell lysate at 20 µg

Lane 6:

MDA-MB-231 (Human breast adenocarcinoma epithelial cell) whole cell lysate at 20 µg

Lane 7:

SK-BR-3 (Human breast adenocarcinoma epithelial cell) whole cell lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/20000 dilution

Predicted band size: 29 kDa

false

• WB

Supplier Data

Western blot - Anti-Histone H3 antibody [EPR16987] - Nuclear Marker and ChIP Grade (AB176842)

Blocking and diluting buffer and concentration : 5% NFDM/TBST Negative control : HepG2 (PMID : 30790340) Lysates were freshly made and used for Western blotting immediately to minimize protein degradation. Exposure time : 125 seconds

All lanes:

Western blot - Anti-ETS1 antibody [EPR27353-81] (<a href='/products/primary-antibodies/ets1-antibody-epr27353-81-ab307672'>ab307672</a>) at 1/1000 dilution

Lane 1:

Daudi (human Burkitt’s lymphoma lymphoblast) whole cell lysate at 20 µg

Lane 2:

HepG2 (human hepatocellar carcinoma epithelial cell)whole cell lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/100000 dilution

Observed band size: 51 kDa

false

Exposure time: 125s

• WB

Supplier Data

Western blot - Anti-Histone H3 antibody [EPR16987] - Nuclear Marker and ChIP Grade (AB176842)

Blocking and diluting buffer and concentration : 5% NFDM/TBST Low expression : liver(PMID : 8521730). In Western blot, anti- H3 antibody (ab176842) loading control staining at 1/100000 dilution. Exposure time : 6 seconds

All lanes:

Western blot - Anti-eIF4A2 antibody [EPR27347-74] (<a href='/products/primary-antibodies/eif4a2-antibody-epr27347-74-ab307728'>ab307728</a>) at 1/1000 dilution

Lane 1:

Human brain tissue lysate at 20 µg

Lane 2:

Human heart tissue lysate at 20 µg

Lane 3:

Human liver tissue lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/100000 dilution

Observed band size: 48 kDa

false

Exposure time: 6s

• WB

Supplier Data

Western blot - Anti-Histone H3 antibody [EPR16987] - Nuclear Marker and ChIP Grade (AB176842)

Low expression : HEK-293.

The expression profile/ molecular weight observed is consistent with what has been described in the literature (PMID : 17431130 and 29296779).

In Western blot, Anti-Histone H3 antibody [EPR16987] - Nuclear Marker and ChIP Grade (ab176842) staining at 1/100000 dilution.

All lanes:

Western blot - Anti-RUNX1 / AML1 antibody [RM1089] (<a href='/products/primary-antibodies/runx1-aml1-antibody-rm1089-ab315398'>ab315398</a>) at 1/1000 dilution

Lane 1:

Jurkat (human T cell leukemia T lymphocyte from peripheral blood) whole cell lysate at 20 µg

Lane 2:

MOLT-4 (human lymphoblastic leukemia T lymphoblast) whole cell lysate at 20 µg

Lane 3:

HEK-293 (human embryonic kidney epithelial cell) whole cell lysate at 20 µg

Lane 4:

THP-1 (human monocytic leukemia monocyte) whole cell lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/100000 dilution

Observed band size: 27-55 kDa,15 kDa

false

Exposure time: 180s

• WB

Supplier Data

Western blot - Anti-Histone H3 antibody [EPR16987] - Nuclear Marker and ChIP Grade (AB176842)

Blocking and diluting buffer and concentration : 5% NFDM/TBST.

Low expression : huh7.

The molecular weight observed is consistent with what has been described in the literature (PMID : 9525922).

In Western blot, Anti-Histone H3 antibody [EPR16987] - Nuclear Marker and ChIP Grade (ab176842) staining at 1/100000 dilution.

All lanes:

Western blot - Anti-Histidine decarboxylase antibody [EPR26392-79] (<a href='/products/primary-antibodies/histidine-decarboxylase-antibody-epr26392-79-ab315444'>ab315444</a>) at 1/1000 dilution

Lane 1:

KU812 (human Peripheral blood basophil) whole cell lysate at 80 µg

Lane 2:

SR (human pleural effusion lymphoblast) whole cell lysate at 80 µg

Lane 3:

Huh7 (human hepatocellar carcinoma epithelial cell) whole cell lysate at 80 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG (HRP) with minimal cross-reactivity with human IgG at 1/2000 dilution

Observed band size: 53 kDa,74 kDa,15 kDa

false

Exposure time: 81s

• WB

Supplier Data

Western blot - Anti-Histone H3 antibody [EPR16987] - Nuclear Marker and ChIP Grade (AB176842)

In Western blot, ab315464 was shown to bind specifically to KMT2A. A band was observed at 180 kDa in wild-type HAP1 cell lysates whereas no signal observed at this size in KMT2A knockout cell line.

Blocking and diluting buffer and concentration : 5% NFDM/TBST.

This antibody has cross reaction with human KMT2B, band at 80 kDa represents KMT2B C-terminal protein.

Lysates were freshly made and used immediately to minimize protein degradation.

There is an unknown band at ~20 kDa.

In Western blot, Anti-Histone H3 antibody [EPR16987] - Nuclear Marker and ChIP Grade (ab176842) staining at 1/100000 dilution.

All lanes:

Western blot - Anti-KMT2A / MLL + KMT2B / MLL4 antibody [EPR28282-16] - C-terminal (<a href='/products/primary-antibodies/kmt2a-mll-kmt2b-mll4-antibody-epr28282-16-c-terminal-ab315464'>ab315464</a>) at 1/1000 dilution

Lane 1:

Wild-type HAP1 (human chronic myelogenous leukemia near-haploid cell) whole cell lysate at 20 µg

Lane 2:

KMT2A knockout HAP1 whole cell lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/100000 dilution

Observed band size: 180 kDa,80 kDa,15 kDa

false

Exposure time: 81s

• WB

Supplier Data

Western blot - Anti-Histone H3 antibody [EPR16987] - Nuclear Marker and ChIP Grade (AB176842)

All lanes:

Western blot - Anti-RBCK1 antibody [EPR28157-53] (<a href='/products/primary-antibodies/rbck1-antibody-epr28157-53-ab309104'>ab309104</a>) at 1/1000 dilution

Lane 1:

HepG2 (human hepatocellar carcinoma epithelial cell) whole cell lysate at 20 µg

Lane 2:

HepG2 transfected with siRNA specifically targeti RBCK1 whole cell lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/100000 dilution

Observed band size: 57 kDa

false

Exposure time: 59s

• WB

Supplier Data

Western blot - Anti-Histone H3 antibody [EPR16987] - Nuclear Marker and ChIP Grade (AB176842)

Blocking and diluting buffer and concentration : 5% NFDM/TBST.

In Western blot, ab315907 was shown to bind specifically to CD276.

Target of interest was observed at 90 KD in wild-type HEK293T cell lysates with no signal observed at this size in CD276 knockout cell line (lane 2, knockout cell line ab266658/ knockout cell lysate ab257097).

Lysates were freshly made, non-boiled and used immediately to minimize protein degradation.

In Western blot, Anti-Histone H3 antibody [EPR16987] - Nuclear Marker and ChIP Grade (ab176842) staining at 1/100000 dilution.

All lanes:

Western blot - Anti-CD276 antibody [RM2041] (<a href='/products/primary-antibodies/cd276-antibody-rm2041-ab315907'>ab315907</a>) at 1/1000 dilution

Lane 1:

Wild-type HEK293T (human embryonic kidney epithelial cell) whole cell lysate at 20 µg

Lane 2:

CD276 knockout HEK293T whole cell lysate at 20 µg

Lane 3:

LNCaP (human prostate carcinoma epithelial cell) whole cell lysate at 20 µg

Lane 4:

Raji (human Burkitts lymphoma B lymphocyte) whole cell lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/100000 dilution

Observed band size: 90 kDa,15 kDa

false

Exposure time: 37s

• WB

Supplier Data

Western blot - Anti-Histone H3 antibody [EPR16987] - Nuclear Marker and ChIP Grade (AB176842)

Blocking and diluting buffer and concentration : 5% NFDM/TBST. This antibody does not cross-react with human BCAT1. In Western blot, anti-His antibody (ab213204) staining at 1/5000 dilution. In Western blot, anti-H3 antibody (ab176842) loading control staining at 1/100000 dilution. Exposure time : 1 second.

All lanes:

Western blot - Anti-BCAT2 antibody [EPR27488-72] (<a href='/products/primary-antibodies/bcat2-antibody-epr27488-72-ab307833'>ab307833</a>) at 1/1000 dilution

Lane 1:

293T (human embryonic kidney epithelial cell)cells transfected with an empty vector containing a his tag whole cell lysate at 20 µg

Lane 2:

293T cells transfected with a human BCAT2 expression vector containing a his tag whole cell lysate at 20 µg

Lane 3:

293T cells transfected with a human BCAT1 expression vector containing a his tag whole cell lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/20000 dilution

Predicted band size: 44 kDa

Observed band size: 37 kDa

false

Exposure time: 1s

• WB

Lab

Western blot - Anti-Histone H3 antibody [EPR16987] - Nuclear Marker and ChIP Grade (AB176842)

Blocking and diluting buffer and concentration : 5% NFDM/TBST

Anti-Histone H3 antibody [EPR16987] - Nuclear Marker and ChIP Grade (ab176842) was used as a loading control at a 1/100000 dilution.

Negative control : HeLa (PMID : 10492038).

All lanes:

Western blot - Anti-MNDA antibody [253A] (<a href='/products/primary-antibodies/mnda-antibody-253a-ab188566'>ab188566</a>) at 1/1000 dilution

Lane 1:

THP-1 (human monocytic leukemia monocyte) whole cell lysate at 20 µg

Lane 2:

HeLa (human cervical adenocarcinoma epithelial cell) whole cell lysate at 20 µg

Secondary

All lanes:

Peroxidase-Conjugated Goat anti-Mouse IgG (H+L) at 1/10000 dilution

Observed band size: 47 kDa

false

Exposure time: 37s

• WB

Supplier Data

Western blot - Anti-Histone H3 antibody [EPR16987] - Nuclear Marker and ChIP Grade (AB176842)

Blocking and diluting buffer and concentration : 5% NFDM/TBST,

In Western blot, Anti-Histone H3 antibody [EPR16987] - Nuclear Marker and ChIP Grade (ab176842) staining at 1/100000 dilution.

All lanes:

Western blot - Anti-VAPB antibody [EPR27026-50] (<a href='/products/primary-antibodies/vapb-antibody-epr27026-50-ab315013'>ab315013</a>) at 1/1000 dilution

Lane 1:

HeLa (human cervical adenocarcinoma epithelial cell) transfected with scrambled siRNA control whole cell lysate at 20 µg

Lane 2:

HeLa transfected with siRNA specifically targeting VAPB whole cell lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/100000 dilution

Observed band size: 27 kDa,15 kDa

false

Exposure time: 15s

• WB

Supplier Data

Western blot - Anti-Histone H3 antibody [EPR16987] - Nuclear Marker and ChIP Grade (AB176842)

Blocking and diluting buffer and concentration : 5% NFDM/TBST Low expression : lung (PMID : 8955374) In Western blot, Anti-Histone H3 antibody [EPR16987] - Nuclear Marker and ChIP Grade (ab176842) staining at 1/100000 dilution. Exposure time : 15 seconds

All lanes:

Western blot - Anti-Calcium channel L type DHPR alpha 2 subunit/CACNA2D1 antibody [EPR23267-59] (<a href='/products/primary-antibodies/calcium-channel-l-type-dhpr-alpha-2-subunit-cacna2d1-antibody-epr23267-59-ab313763'>ab313763</a>) at 1/1000 dilution

Lane 1:

Human skeletal muscle tissue lysate at 20 µg

Lane 2:

Human lung tissue lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/100000 dilution

Observed band size: 150 kDa

false

Exposure time: 15s

• WB

Supplier Data

Western blot - Anti-Histone H3 antibody [EPR16987] - Nuclear Marker and ChIP Grade (AB176842)

Blocking and diluting buffer and concentration : 5% NFDM/TBST

Negative control : K562(PMID : 35134130)

The identity of the bands between 75 kDa and 25 kDa are unknown.

In Western blot, Anti-Histone H3 antibody [EPR16987] - Nuclear Marker and ChIP Grade (ab176842) staining at 1/100000 dilution.

All lanes:

Western blot - Anti-SEMA4A antibody [EPR27097-20] (<a href='/products/primary-antibodies/sema4a-antibody-epr27097-20-ab314648'>ab314648</a>) at 1/1000 dilution

Lane 1:

THP-1(human monocytic leukemia monocyte) membrane fraction at 50 µg

Lane 2:

THP-1 non-membrane fraction at 50 µg

Lane 3:

THP-1(human monocytic leukemia monocyte) whole cell lysate at 50 µg

Lane 4:

K562 (human chronic myelogenous leukemia lymphoblast) whole cell lysate at 50 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG (HRP) with minimal cross-reactivity with human IgG at 1/5000 dilution

Observed band size: 84 kDa,15 kDa

false

Exposure time: 180s

• WB

Supplier Data

Western blot - Anti-Histone H3 antibody [EPR16987] - Nuclear Marker and ChIP Grade (AB176842)

Blocking and diluting buffer and concentration : 5% NFDM/TBST.

The identity of the higher MW band at approximately 50 kDa (in lane 2) is unknown.

Samples are non-boiled as boiling may cause protein aggregation.

In Western blot, Anti-Histone H3 antibody [EPR16987] - Nuclear Marker and ChIP Grade (ab176842) staining at 1/100000 dilution.

In Western blot, Anti-Integrin alpha V antibody [EPR16800] (ab179475) staining at 1/5000 dilution.

All lanes:

Western blot - Anti-GPCR GPRC5D antibody [EPR28376-41] (<a href='/products/primary-antibodies/gpcr-gprc5d-antibody-epr28376-41-ab315808'>ab315808</a>) at 1/1000 dilution

Lane 1:

RPMI-8226 (human plasmacytoma, myeloma B lymphocyte) membrane fraction at 50 µg

Lane 2:

RPMI-8226 nuclear fraction at 50 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/20000 dilution

Observed band size: 33 kDa,125 kDa,15 kDa

false

Exposure time: 15s

• WB

Supplier Data

Western blot - Anti-Histone H3 antibody [EPR16987] - Nuclear Marker and ChIP Grade (AB176842)

ErbB2 / HER2 (phospho Y1221 + Y1222) expression can be induced by EGF treatment (PMID : 7728765).

In Western blot, Anti-Histone H3 antibody [EPR16987] - Nuclear Marker and ChIP Grade (ab176842) staining at 1/100000 dilution.

In Western blot, Anti-ErbB2 / HER2 antibody - Total protein control (ab214275) staining at 1/1000 dilution.

All lanes:

Western blot - Anti-ErbB2 / HER2 (phospho Y1221 + Y1222) antibody [EPR28391-67] (<a href='/products/primary-antibodies/erbb2-her2-phospho-y1221-y1222-antibody-epr28391-67-ab316758'>ab316758</a>) at 1/1000 dilution

Lane 1:

Untreated HeLa (human cervix adenocarcinoma epithelial cell) starved for 4 hours whole cell lysate (untreated membrane) at 20 µg with 5% NFDM/TBST

Lane 2:

HeLa starved for 4 hours, then treated with /ml EGF for 15 minutes whole cell lysate (untreated membrane) at 20 µg with 5% NFDM/TBST

Lane 3:

HeLa starved for 4 hours, then treated with /ml EGF for 15 minutes whole cell lysate (alkaline phosphatase treated membrane) at 20 µg with 5% NFDM/TBST

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/100000 dilution

Observed band size: 180 kDa,15 kDa

false

Exposure time: 81s

• WB

Lab

Western blot - Anti-Histone H3 antibody [EPR16987] - Nuclear Marker and ChIP Grade (AB176842)

Blocking buffer and concentration : 5% NFDM/TBST  Diluting buffer and concentration : 5% NFDM/TBST ab176323 detects a higher band in nuclear fraction than in non-nuclear fraction, which might be the correct band of interest. ab213204, ab176842 and ab181602 were used as loading controls.

All lanes:

Western blot - Anti-LXR alpha antibody [EPR6508(N)] (<a href='/products/primary-antibodies/lxr-alpha-antibody-epr6508n-ab176323'>ab176323</a>) at 1/1000 dilution

Lane 1:

HEK 293 transfected with blank vectors, whole cell lysate at 20 µg

Lane 2:

HEK 293 transfected with blank vectors, non-nuclear fraction at 20 µg

Lane 3:

HEK 293 transfected with blank vectors, nuclear fraction at 20 µg

Lane 4:

HEK 293 transfected with His tagged human LXR alpha full length expression vectors, non-nuclear fraction at 20 µg

Lane 5:

HEK 293 transfected with His tagged human LXR alpha full length expression vectors, nuclear fraction at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/20000 dilution

Predicted band size: 50 kDa

Observed band size: 50 kDa

false

Exposure time: 10s

• WB

Supplier Data

Western blot - Anti-Histone H3 antibody [EPR16987] - Nuclear Marker and ChIP Grade (AB176842)

Blocking and diluting buffer and concentration : 5% NFDM/TBST.

Negative control : Hela (PMID : 35164755).

Samples are non-boiled as boiling may cause protein aggregation.

The molecular weight observed is consistent with what has been described in the literature (PMID : 36631513).

In Western blot, anti- H3 antibody (ab176842) loading control staining at 1/100000 dilution.

All lanes:

Western blot - Anti-VE Cadherin antibody [EPR27436-55] (<a href='/products/primary-antibodies/ve-cadherin-antibody-epr27436-55-ab313632'>ab313632</a>) at 1/1000 dilution

Lane 1:

HUVEC (human umbilical vein endothelial cell) whole cell lysate at 20 µg

Lane 2:

HeLa (human cervix adenocarcinoma epithelial cell) whole cell lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/100000 dilution

Observed band size: 87-100 kDa,130 kDa

false

Exposure time: 3s

• WB

Supplier Data

Western blot - Anti-Histone H3 antibody [EPR16987] - Nuclear Marker and ChIP Grade (AB176842)

Blocking and diluting buffer and concentration : 5% NFDM/TBST

Negative control : kidney (PMID : 11922625).

In Western blot, anti- H3 antibody (ab176842) loading control staining at 1/100000 dilution.

This blot was developed using a high sensitivity ECL substrate. The high-sensitivity ECL substrate used allows for the detection of proteins in the mid-femtogram range.

All lanes:

Western blot - Anti-DDX53/CT26 antibody [EPR27384-107] (<a href='/products/primary-antibodies/ddx53-ct26-antibody-epr27384-107-ab313333'>ab313333</a>) at 1/1000 dilution

Lane 1:

Human testis tissue nuclear fraction at 40 µg

Lane 2:

Human kidney tissue nuclear fraction at 40 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG (HRP) with minimal cross-reactivity with human IgG at 1/2000 dilution

Observed band size: 74 kDa

true

Exposure time: 70s

• WB

Supplier Data

Western blot - Anti-Histone H3 antibody [EPR16987] - Nuclear Marker and ChIP Grade (AB176842)

In Western blot, Anti-Histone H3 antibody [EPR16987] - Nuclear Marker and ChIP Grade (ab176842) staining at 1/100000 dilution.

In Western blot, Anti-ErbB2 / HER2 antibody - Total protein control (ab214275) staining at 1/1000 dilution.

All lanes:

Western blot - Anti-ErbB2 / HER2 (phospho Y1221 + Y1222) antibody [EPR28391-67] (<a href='/products/primary-antibodies/erbb2-her2-phospho-y1221-y1222-antibody-epr28391-67-ab316758'>ab316758</a>) at 1/1000 dilution

Lane 1:

Untreated T47D (human ductal breast epithelial tumor epithelial cell) starved for 4 hours whole cell lysate (untreated membrane) at 20 µg with 5% NFDM/TBST

Lane 2:

T47D starved for 4 hours, then treated with /ml Neuregulin for 10 minutes whole cell lysate (untreated membrane) at 20 µg with 5% NFDM/TBST

Lane 3:

T47D starved for 4 hours, then treated with /ml Neuregulin for 10 minutes whole cell lysate (alkaline phosphatase treated membrane) at 20 µg with 5% NFDM/TBST

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/100000 dilution

Observed band size: 180 kDa,15 kDa

false

Exposure time: 103s

• WB

Supplier Data

Western blot - Anti-Histone H3 antibody [EPR16987] - Nuclear Marker and ChIP Grade (AB176842)

All lanes:

Western blot - Anti-KACL antibody [EPR27403-26] (<a href='/products/primary-antibodies/kacl-antibody-epr27403-26-ab309102'>ab309102</a>) at 1/1000 dilution

Lane 1:

U-937 (human histiocytic lymphoma monocyte) whole cell lysate at 20 µg

Lane 2:

THP-1 (human monocytic leukemia monocyte) whole cell lysate at 20 µg

Lane 3:

HeLa (human cervix adenocarcinoma epithelial cell) whole cell lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/100000 dilution

Observed band size: 32 kDa

false

Exposure time: 15s

• WB

Supplier Data

Western blot - Anti-Histone H3 antibody [EPR16987] - Nuclear Marker and ChIP Grade (AB176842)

Blocking and diluting buffer and concentration : 5% NFDM/TBST.

Negative control : SH-SY5Y (PMID : 30650643 and PMID : 27345400)

In Western blot, Anti-Histone H3 antibody [EPR16987] - Nuclear Marker and ChIP Grade (ab176842) staining at 1/100000 dilution.

All lanes:

Western blot - Anti-Cytokeratin 19 antibody [EP1580Y] - Mouse IgG2a (Chimeric) (<a href='/products/primary-antibodies/cytokeratin-19-antibody-ep1580y-mouse-igg2a-chimeric-ab323562'>ab323562</a>) at 1/1000 dilution

Lane 1:

MCF7 (human breast adenocarcinoma epithelial cell) whole cell lysate at 20 µg

Lane 2:

SH-SY5Y (human neuroblastoma epithelial cell) whole cell lysate at 40 µg

Secondary

All lanes:

Western blot - Goat Anti-Mouse IgG Fc (HRP) preadsorbed (<a href='/products/secondary-antibodies/goat-mouse-igg-fc-hrp-preadsorbed-ab98717'>ab98717</a>) at 1/20000 dilution

Observed band size: 44 kDa,17 kDa

false

Exposure time: 4s

• WB

Supplier Data

Western blot - Anti-Histone H3 antibody [EPR16987] - Nuclear Marker and ChIP Grade (AB176842)

Blocking and diluting buffer and concentration : 5% NFDM/TBST.

In Western blot, Anti-Histone H3 antibody [EPR16987] - Nuclear Marker and ChIP Grade (ab176842) staining at 1/100000 dilution.

All lanes:

Western blot - Anti-SPG3A/ATL1 antibody [EPR28057-64] (<a href='/products/primary-antibodies/spg3a-atl1-antibody-epr28057-64-ab316111'>ab316111</a>) at 1/1000 dilution

Lane 1:

293T (human embryonic kidney epithelial cell) transfected with scrambled siRNA control whole cell lysate at 20 µg

Lane 2:

293T transfected with siRNA specifically targeting ATL1 whole cell lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/100000 dilution

Observed band size: 63 kDa,15 kDa

true

Exposure time: 92s

• WB

Supplier Data

Western blot - Anti-Histone H3 antibody [EPR16987] - Nuclear Marker and ChIP Grade (AB176842)

In Western blot, Anti-Histone H3 antibody [EPR16987] - Nuclear Marker and ChIP Grade (ab176842) staining at 1/100000 dilution.

All lanes:

Western blot - Anti-AIMP2/p38 antibody [EPR29127-48] (<a href='/products/primary-antibodies/aimp2-p38-antibody-epr29127-48-ab316937'>ab316937</a>) at 1/1000 dilution

Lane 1:

HeLa (human cervical adenocarcinoma epithelial cell) transfected with scrambled siRNA control whole cell lysate at 20 µg with 5% NFDM/TBST

Lane 2:

HeLa transfected with siRNA specifically targeting AIMP2 whole cell lysate at 20 µg with 5% NFDM/TBST

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/100000 dilution

Observed band size: 35 kDa,15 kDa

false

Exposure time: 180s

• WB

Supplier Data

Western blot - Anti-Histone H3 antibody [EPR16987] - Nuclear Marker and ChIP Grade (AB176842)

Blocking and diluting buffer and concentration : 5% NFDM/TBST.

Negative control : Raji (PMID : 32808354), K-562 (PMID : 32808354), HeLa.

In Western blot, Anti-Histone H3 antibody [EPR16987] - Nuclear Marker and ChIP Grade (ab176842) staining at 1/100000 dilution.

All lanes:

Western blot - Anti-CD2 antibody [EPR27426-17] (<a href='/products/primary-antibodies/cd2-antibody-epr27426-17-ab314761'>ab314761</a>) at 1/1000 dilution

Lane 1:

Jurkat (human T cell leukemia T lymphocyte from peripheral blood) whole cell lysate at 20 µg

Lane 2:

Raji (human Burkitt's lymphoma B lymphocyte) whole cell lysate at 20 µg

Lane 3:

MOLT-4 (human lymphoblastic leukemia T lymphoblast) whole cell lysate at 20 µg

Lane 4:

No-GFP-CD16. NK-92 (CD16 ectopically expressed natural killer(NK-92®) cell with no GFP tag) whole cell lysate at 20 µg

Lane 5:

HeLa (human cervical adenocarcinoma epithelial cell) whole cell lysate at 20 µg

Lane 6:

K-562 (human chronic myelogenous leukemia lymphoblast) whole cell lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/20000 dilution

Observed band size: 35-60 kDa,15 kDa

false

Exposure time: 48s

• WB

Supplier Data

Western blot - Anti-Histone H3 antibody [EPR16987] - Nuclear Marker and ChIP Grade (AB176842)

Blocking and diluting buffer and concentration : 5% NFDM/TBST. In Western blot, anti- H3 antibody (ab176842) loading control staining at 1/100000 dilution. Exposure time : 70 seconds.

All lanes:

Western blot - Anti-RANTES antibody [EPR26519-16] (<a href='/products/primary-antibodies/rantes-antibody-epr26519-16-ab307712'>ab307712</a>) at 1/1000 dilution

Lane 1:

Untreated THP-1 (human monocytic leukemia monocyte) whole cell lysate at 20 µg

Lane 2:

THP-1 treated with 100ng/ml PMA for 48 hours and 1ug/ml LPS for 8 hours, and 600ng/ml Brefeldin A for the 3 hours whole cell lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/100000 dilution

Predicted band size: 10 kDa

Observed band size: 10 kDa

false

Exposure time: 70s

• WB

Supplier Data

Western blot - Anti-Histone H3 antibody [EPR16987] - Nuclear Marker and ChIP Grade (AB176842)

Negative control : Jurkat

Low expression : liver

In Western blot, Anti-Histone H3 antibody [EPR16987] - Nuclear Marker and ChIP Grade (ab176842) staining at 1/100000 dilution.

Exposure time : Lane1-2 : 15seconds, lane3-5 : 180 seconds

All lanes:

Western blot - Anti-MNDA antibody [EPR28267-44] (<a href='/products/primary-antibodies/mnda-antibody-epr28267-44-ab316967'>ab316967</a>) at 1/1000 dilution

Lane 1:

U-937 (human histiocytic lymphoma monocyte) whole cell lysate at 20 µg with NFDM/TBST

Lane 2:

Jurkat (human t cell leukemia t lymphocyte from peripheral blood) whole cell lysate at 20 µg with NFDM/TBST

Lane 3:

Human spleen tissue lysate at 20 µg with NFDM/TBST

Lane 4:

Human tonsil tissue lysate at 20 µg with NFDM/TBST

Lane 5:

Human liver tissue lysate at 20 µg with NFDM/TBST

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/100000 dilution

Observed band size: 27 kDa,15 kDa

false

• WB

Supplier Data

Western blot - Anti-Histone H3 antibody [EPR16987] - Nuclear Marker and ChIP Grade (AB176842)

Blocking and diluting buffer and concentration : 5% NFDM/TBST.

Low expression : HT-29 (PMID : 31720873)

To minimize protein degradation, cells were lysed immediately after harvest and then applied to a gel and transfer membrane for Western blotting as soon as possible.

The identity of the band higher than 96 kDa is unknown.

In Western blot, Anti-Histone H3 antibody [EPR16987] - Nuclear Marker and ChIP Grade (ab176842) staining at 1/100000 dilution.

All lanes:

Western blot - Anti-TTK/Mps1 antibody [EPR29753-583] (<a href='/products/primary-antibodies/ttk-mps1-antibody-epr29753-583-ab323713'>ab323713</a>) at 1/1000 dilution

Lane 1:

SW480 (human colorectal adenocarcinoma epithelial cell) whole cell lysate at 20 µg

Lane 2:

HT-29 (human colorectal adenocarcinoma epithelial cell) whole cell lysate at 20 µg

Lane 3:

PC-3 (human prostate adenocarcinoma epithelial cell) whole cell lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/20000 dilution

Observed band size: 97 kDa,15 kDa

false

Exposure time: 26s