Anti-ETS2 抗体 [EPR22419]

• IP

Supplier Data

Immunoprecipitation - Anti-ETS2 antibody [EPR22419] (AB219948)

ETS2 was immunoprecipitated from 0.35 mg K562 (human chronic myelogenous leukemia cell line from bone marrow ) whole cell lysate with ab219948 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab219948 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/5000 dilution.

Lane 1 : K562 whole cell lysate 10 μg (Input).
Lane 2 : ab219948 IP in K562 whole cell lysate.
Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab219948 in K562 whole cell lysate.

Blocking/Dilution buffer : 5% NFDM/TBST.
Exposure time : 3 minutes.

The two immunoprecipitated bands represent non-phosphorylated and phosphorylated ETS2.

All lanes:

Immunoprecipitation - Anti-ETS2 antibody [EPR22419] (ab219948)

Predicted band size: 53 kDa

Observed band size: 53 kDa

false

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-ETS2 antibody [EPR22419] (AB219948)

Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized RAW 264.7 (mouse macrophage cell line transformed with Abelson murine leukemia virus) cells labeling ETS2 with ab219948 at 1/500 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing nuclear staining in RAW 264.7 cells treated with lipopolysaccharide (500 ng/ml) for 6 hours. The nuclear counterstain is DAPI (blue).
Counterstained with ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor^® 594) at a 1/200 dilution (red).
Secondary antibody contril : Used PBS instead of primary antibody, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) (ab150077) secondary antibody at 1/1000 dilution.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ETS2 antibody [EPR22419] (AB219948)

Immunohistochemical analysis of paraffin-embedded rat thymus tissue labeling ETS2 with ab219948 at 1/500 dilution, followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP). Nuclear staining on rat thymus (PMID : 3472202, PMID : 26926107) is observed. Counterstained with hematoxylin.
Secondary antibody only control : Used PBS instead of primary antibody, secondary antibody is a ready to use Goat Anti-Rabbit IgG H&L (HRP).
Perform heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0).

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ETS2 antibody [EPR22419] (AB219948)

Immunohistochemical analysis of paraffin-embedded rat cerebellum tissue labeling ETS2 with ab219948 at 1/500 dilution, followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP).

Negative control : No staining on rat cerebellum (PMID : 3472202) is observed. Counterstained with hematoxylin.

Perform heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0).

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ETS2 antibody [EPR22419] (AB219948)

Immunohistochemical analysis of paraffin-embedded mouse thymus tissue labeling ETS2 with ab219948 at 1/500 dilution, followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP). Nuclear staining on mouse thymus (PMID : 3472202, PMID : 26926107) is observed. Counterstained with hematoxylin.
Secondary antibody only control : Used PBS instead of primary antibody, secondary antibody is a ready to use Goat Anti-Rabbit IgG H&L (HRP).
Perform heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0).

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ETS2 antibody [EPR22419] (AB219948)

Immunohistochemical analysis of paraffin-embedded mouse cerebrum tissue labeling ETS2 with ab219948 at 1/500 dilution, followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP).

Negative control : No staining on mouse cerebrum (PMID : 3472202) is observed. Counterstained with hematoxylin.

Perform heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0).

• WB

Supplier Data

Western blot - Anti-ETS2 antibody [EPR22419] (AB219948)

Blocking/Dilution buffer : 5% NFDM/TBST.

Exposure times : Lane 1 : 26 seconds, Lane 2 : 3 minutes.

Lane 1 was developed using a higher sensitivity ECL substrate.

All lanes:

Western blot - Anti-ETS2 antibody [EPR22419] (ab219948) at 1/1000 dilution

Lane 1:

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

Lane 2:

Human thymus at 10 µg

Secondary

Lane 1:

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

Lane 2:

Western blot - VeriBlot for IP Detection Reagent (HRP) (<a href='/products/reagents/veriblot-for-ip-detection-reagent-hrp-ab131366'>ab131366</a>) at 1/1000 dilution

Predicted band size: 53 kDa

Observed band size: 53 kDa

false

• WB

Lab

Western blot - Anti-ETS2 antibody [EPR22419] (AB219948)

Western blot : Recombinant Anti-ETS2 antibody [EPR22419] ab219948 staining at 1/1000 dilution, shown in green; Mouse anti-CANX ab238078 loading control staining at 1/20000 dilution, shown in magenta. A band was observed at 53 kDa. To generate this image, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 5pc Milk in TBS-0.1 % Tween^® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit 800CW and Goat anti-Mouse 680RD at 1/20,000 dilution.

All lanes:

Western blot - Anti-ETS2 antibody [EPR22419] (ab219948) at 1/1000 dilution

Lane 1:

Wild-type THP-1 Nuclear Fraction at 40 µg

Lane 2:

Western blot - Human ETS2 knockout THP1 cell line (<a href='/products/cell-lines/human-ets2-knockout-thp1-cell-line-ab290239'>ab290239</a>) at 40 kDa

Lane 3:

K562 Nuclear Fraction at 20 µg

Lane 4:

T-47D Nuclear Fraction at 20 µg

Secondary

All lanes:

Goat anti-Rabbit 800CW & Goat anti-Mouse 680RD at 1/20000 dilution

Predicted band size: 53 kDa

Observed band size: 53 kDa

false

• WB

Lab

Western blot - Anti-ETS2 antibody [EPR22419] (AB219948)

Western blot : Recombinant Anti-ETS2 antibody [EPR22419] ab219948 staining at 1/1000 dilution, shown in green; Mouse anti-CANX ab238078 loading control staining at 1/20000 dilution, shown in magenta. A band was observed at 53 kDa in Wild-type THP-1 Nuclear cell lysates with no signal observed at this size in ETS2 knockout THP-1 Nuclear cell line. To generate this image, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 5pc Milk in TBS-0.1 % Tween^® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit 800CW and Goat anti-Mouse 680RD at 1/20,000 dilution.

All lanes:

Western blot - Anti-ETS2 antibody [EPR22419] (ab219948) at 1/1000 dilution

Lane 1:

Wild-type THP-1 Nuclear at 40 µg

Lane 2:

Western blot - Human ETS2 knockout THP1 cell line (<a href='/products/cell-lines/human-ets2-knockout-thp1-cell-line-ab290239'>ab290239</a>) at 40 µg

Lane 3:

K-562 Nuclear at 20 µg

Lane 4:

T-47D Nuclear at 20 µg

Secondary

All lanes:

Goat anti-Rabbit 800CW & Goat anti-Mouse 680RD at 1/20000 dilution

Predicted band size: 53 kDa

Observed band size: 53 kDa,77 kDa

false

• WB

Supplier Data

Western blot - Anti-ETS2 antibody [EPR22419] (AB219948)

Blocking/Dilution buffer : 5% NFDM/TBST.

The expression profile observed is consistent with what has been described in the literature (PMID : 15590657). The doublets represent ETS2 and its phosphorylated form.

This blot was developed using a higher sensitivity ECL substrate.

All lanes:

Western blot - Anti-ETS2 antibody [EPR22419] (ab219948) at 1/1000 dilution

Lane 1:

Untreated RAW 264.7 (mouse Abelson murine leukemia virus-induced tumor macrophage), whole cell lysate at 20 µg

Lane 2:

RAW 264.7 treated with 500ng/ml lipopolysaccharides(LPS) for 6 hours 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: 53 kDa

Observed band size: 50-55 kDa

false

Exposure time: 5s