Name: Anti-NFkB p105 / p50 antibody [E381]
Brand: RabMAb
SKU: ab32360
Rating: 4 (21 reviews)

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Western blot - Anti-NFkB p105 / p50 antibody [E381] (ab32360)

Key features and details

Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
Rabbit monoclonal [E381] to NFkB p105 / p50
Suitable for: WB, IHC-P
Knockout validated
Reacts with: Mouse, Rat, Human

Alexa Fluor® 555 Alexa Fluor® 568 Alexa Fluor® 594 Alexa Fluor® 647 Alexa Fluor® 750 Carrier Free HRP

产品名称

Anti-NFkB p105 / p50抗体[E381]
参阅全部 NFkB p105 / p50 一抗
描述

兔单克隆抗体[E381] to NFkB p105 / p50
宿主

Rabbit
特异性

This antibody will detect both forms: p50 and p105.
经测试应用

适用于: WB, IHC-Pmore details
不适用于: Flow Cyt or IP
种属反应性

与反应: Mouse, Rat, Human
免疫原

Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
阳性对照
- WB: HeLa, MCF-7 and PC-12 cell lysates. IHC-P: human prostate carcinoma and bladder carcinoma tissues.
常规说明
This product is a recombinant monoclonal antibody, which offers several advantages including:
- - High batch-to-batch consistency and reproducibility
- - Improved sensitivity and specificity
- - Long-term security of supply
- - Animal-free production
For more information see here.
Our RabMAb^® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb^® patents.

形式

Liquid
存放说明

Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C. Avoid freeze / thaw cycle.
存储溶液

pH: 7.20
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol, 0.05% BSA
Concentration information loading...
纯度

Protein A purified
克隆

单克隆
克隆编号

E381
同种型

IgG
研究领域
- Cell Biology
- Apoptosis
- Intracellular
- NFkB
- p50
- Cancer
- Cell Death
- Apoptosis
- Apoptosis Markers
- NFkB
- p50

Alternative Versions
Assay kits
- Human NFkB p50 Transcription Factor Assay Kit (ab133105)
Compatible Secondaries
Isotype control
- Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)
Positive Controls
- HeLa whole cell lysate (ab150035)
- HeLa whole cell lysate (ab29545)

The Abpromise guarantee

Abpromise™承诺保证使用ab32360于以下的经测试应用

“应用说明”部分下显示的仅为推荐的起始稀释度；实际最佳的稀释度/浓度应由使用者检定。

应用	Ab评论	说明
WB	(11)	1/1000 - 1/10000. Detects a band of approximately 50, 105 kDa (predicted molecular weight: 50 kDa).
IHC-P	(5)	1/250 - 1/500. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. See IHC antigen retrieval protocols.

说明
WB 1/1000 - 1/10000. Detects a band of approximately 50, 105 kDa (predicted molecular weight: 50 kDa).
IHC-P 1/250 - 1/500. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. See IHC antigen retrieval protocols.

应用说明

Is unsuitable for Flow Cyt or IP.

功能

NF-kappa-B is a pleiotropic transcription factor which is present in almost all cell types and is involved in many biological processed such as inflammation, immunity, differentiation, cell growth, tumorigenesis and apoptosis. NF-kappa-B is a homo- or heterodimeric complex formed by the Rel-like domain-containing proteins RELA/p65, RELB, NFKB1/p105, NFKB1/p50, REL and NFKB2/p52 and the heterodimeric p65-p50 complex appears to be most abundant one. The dimers bind at kappa-B sites in the DNA of their target genes and the individual dimers have distinct preferences for different kappa-B sites that they can bind with distinguishable affinity and specificity. Different dimer combinations act as transcriptional activators or repressors, respectively. NF-kappa-B is controlled by various mechanisms of post-translational modification and subcellular compartmentalization as well as by interactions with other cofactors or corepressors. NF-kappa-B complexes are held in the cytoplasm in an inactive state complexed with members of the NF-kappa-B inhibitor (I-kappa-B) family. In a conventional activation pathway, I-kappa-B is phosphorylated by I-kappa-B kinases (IKKs) in response to different activators, subsequently degraded thus liberating the active NF-kappa-B complex which translocates to the nucleus. NF-kappa-B heterodimeric p65-p50 and RelB-p50 complexes are transcriptional activators. The NF-kappa-B p50-p50 homodimer is a transcriptional repressor, but can act as a transcriptional activator when associated with BCL3. NFKB1 appears to have dual functions such as cytoplasmic retention of attached NF-kappa-B proteins by p105 and generation of p50 by a cotranslational processing. The proteasome-mediated process ensures the production of both p50 and p105 and preserves their independent function, although processing of NFKB1/p105 also appears to occur post-translationally. p50 binds to the kappa-B consensus sequence 5'-GGRNNYYCC-3', located in the enhancer region of genes involved in immune response and acute phase reactions. In a complex with MAP3K8, NFKB1/p105 represses MAP3K8-induced MAPK signaling; active MAP3K8 is released by proteasome-dependent degradation of NFKB1/p105.
序列相似性

Contains 7 ANK repeats.
Contains 1 death domain.
Contains 1 RHD (Rel-like) domain.
结构域

The C-terminus of p105 might be involved in cytoplasmic retention, inhibition of DNA-binding, and transcription activation.
Glycine-rich region (GRR) appears to be a critical element in the generation of p50.
翻译后修饰

While translation occurs, the particular unfolded structure after the GRR repeat promotes the generation of p50 making it an acceptable substrate for the proteasome. This process is known as cotranslational processing. The processed form is active and the unprocessed form acts as an inhibitor (I kappa B-like), being able to form cytosolic complexes with NF-kappa B, trapping it in the cytoplasm. Complete folding of the region downstream of the GRR repeat precludes processing.
Phosphorylation at 'Ser-903' and 'Ser-907' primes p105 for proteolytic processing in response to TNF-alpha stimulation. Phosphorylation at 'Ser-927' and 'Ser-932' are required for BTRC/BTRCP-mediated proteolysis.
Polyubiquitination seems to allow p105 processing.
S-nitrosylation of Cys-61 affects DNA binding.
细胞定位

Nucleus. Cytoplasm. Nuclear, but also found in the cytoplasm in an inactive form complexed to an inhibitor.
Target information above from: UniProt accession P19838 The UniProt Consortium
The Universal Protein Resource (UniProt) in 2010
Nucleic Acids Res. 38:D142-D148 (2010) .

Information by UniProt
数据库链接
- Entrez Gene: 4790 Human
- Entrez Gene: 18033 Mouse
- Entrez Gene: 81736 Rat
- Omim: 164011 Human
- SwissProt: P19838 Human
- SwissProt: P25799 Mouse
- SwissProt: Q63369 Rat
- Unigene: 618430 Human
- Unigene: 256765 Mouse
- Unigene: 2411 Rat
see all
别名
- DKFZp686C01211 antibody
- DNA binding factor KBF1 antibody
- DNA binding factor KBF1 EBP1 antibody
- DNA-binding factor KBF1 antibody
- EBP 1 antibody
- EBP-1 antibody
- EBP1 antibody
- KBF1 antibody
- MGC54151 antibody
- nf b antibody
- NF kappa B antibody
- NF kappaB antibody
- NF kappabeta antibody
- NF kB1 antibody
- NF-kappaB antibody
- NFkappaB antibody
- NFKB 1 antibody
- NFKB p105 antibody
- NFKB p50 antibody
- NFKB-p105 antibody
- Nfkb1 antibody
- NFKB1_HUMAN antibody
- Nuclear factor kappa B DNA binding subunit antibody
- Nuclear factor kappa-B antibody
- Nuclear factor kappa-B, subunit 1 antibody
- Nuclear factor NF kappa B p105 subunit antibody
- Nuclear factor NF kappa B p50 subunit antibody
- Nuclear factor NF-kappa-B p50 subunit antibody
- Nuclear factor of kappa light chain gene enhancer in B cells 1 antibody
- Nuclear factor of kappa light polypeptide gene enhancer in B cells 1 antibody
- Nuclear factor of kappa light polypeptide gene enhancer in B-cells 1 antibody
- p105 antibody
- p50 antibody
- p84/NF-kappa-B1 p98 antibody
- Transcription factor NFKB1 antibody
see all

Western blot - Anti-NFkB p105 / p50 antibody [E381] (ab32360)This image is courtesy of an anonymous Abreview

Anti-NFkB p105 / p50 antibody [E381] (ab32360) at 1/1000 dilution + mouse neural stem cell lysate at 40 µg

Secondary
Alexa Fluor 790 Goat fluorescent secondary at 1/10000 dilution

Performed under reducing conditions.

Predicted band size: 50 kDa
Observed band size: 100,50 kDa why is the actual band size different from the predicted?

Exposure time: 1 hour
See Abreview
Western blot - Anti-NFkB p105 / p50 antibody [E381] (ab32360)

All lanes : Anti-NFkB p105 / p50 antibody [E381] (ab32360)

Lane 1 : Wild-type HAP1 cell lysate
Lane 2 : NFkB p105 / p50 knockout HAP1 cell lysate
Lane 3 : HeLa cell lysate
Lane 4 : Jurkat cell lysate

Lysates/proteins at 20 µg per lane.

Predicted band size: 50 kDa

Lanes 1 - 4: Merged signal (red and green). Green - ab32360 observed at 120, 50 kDa. Red - loading control, ab8245, observed at 37 kDa.

ab32360 was shown to specifically react with NFkB p105 / p50 when NFkB p105 / p50 knockout samples were used. Wild-type and NFkB p105 / p50 knockout samples were subjected to SDS-PAGE. ab32360 and ab8245 (loading control to GAPDH) were diluted at 1/1000 and 1/10 000 respectively and and incubated overnight at 4°C. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye^® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preadsorbed (ab216776) secondary antibodies at 1/10000 dilution for 1 hour at room temperature before imaging.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-NFkB p105 / p50 antibody [E381] (ab32360)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human bladder carcinoma tissue labelling NFkB p105 / p50 with purified ab32360 at 1/250. Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9. ab97051, a HRP-conjugated goat anti-rabbit IgG (H+L) was used as the secondary antibody (1/500). Negative control using PBS instead of primary antibody. Counterstained with hematoxylin.
Western blot - Anti-NFkB p105 / p50 antibody [E381] (ab32360)

All lanes : Anti-NFkB p105 / p50 antibody [E381] (ab32360) at 1/1000 dilution

Lane 1 : Wild-type HeLa cell lysate
Lane 2 : NFKB1 CRISPR/Cas9 edited HeLa cell lysate

Lysates/proteins at 20 µg per lane.

Performed under reducing conditions.

Predicted band size: 50 kDa
Observed band size: 105 kDa why is the actual band size different from the predicted?

Lanes 1- 2: Merged signal (red and green). Green - ab32360 observed at 105 kDa. Red - Anti-GAPDH antibody [6C5] - Loading Control (ab8245) observed at 37 kDa.

ab32360 was shown to react with NFkB p105 / p50 in wild-type HeLa cells in western blot. The band observed in CRISPR/Cas9 edited cell line ab264823 (CRISPR/Cas9 edited cell lysate ab257003) lane below 105kDa may represent truncated forms and cleaved fragments. This has not been investigated further. Wild-type HeLa and NFKB1 CRISPR/Cas9 HeLa HeLa cell lysates were subjected to SDS-PAGE. Membrane was blocked for 1 hour at room temperature in 0.1% TBST with 3% non-fat dried milk. ab32360 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C at a 1 in 1000 dilution and a 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye^®800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye^®680RD) preadsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.
Western blot - Anti-NFkB p105 / p50 antibody [E381] (ab32360)

Anti-NFkB p105 / p50 antibody [E381] (ab32360) at 1/10000 dilution (purified) + HeLa cell lysate at 20 µg

Secondary
Peroxidase conjugated goat anti-rabbit IgG (H+L) at 1/1000 dilution

Predicted band size: 50 kDa
Observed band size: 105,50 kDa why is the actual band size different from the predicted?

Blocking and dilution buffer: 5% NFDM/TBST.
Western blot - Anti-NFkB p105 / p50 antibody [E381] (ab32360)

Anti-NFkB p105 / p50 antibody [E381] (ab32360) at 1/50000 dilution (purified) + MCF-7 cell lysate at 10 µg

Secondary
Peroxidase conjugated goat anti-rabbit IgG (H+L) at 1/1000 dilution

Predicted band size: 50 kDa
Observed band size: 105 kDa why is the actual band size different from the predicted?
Additional bands at: 50 kDa (possible non-specific binding)

Blocking and dilution buffer: 5% NFDM/TBST.
Western blot - Anti-NFkB p105 / p50 antibody [E381] (ab32360)

All lanes : Anti-NFkB p105 / p50 antibody [E381] (ab32360) at 1/10000 dilution (purified)

Lane 1 : PC-12 cell lysate
Lane 2 : NIH/3T3 cell lysate

Lysates/proteins at 20 µg per lane.

Secondary
All lanes : Peroxidase conjugated goat anti-rabbit IgG (H+L) at 1/1000 dilution

Predicted band size: 50 kDa
Observed band size: 105 kDa why is the actual band size different from the predicted?
Additional bands at: 50 kDa (possible non-specific binding)

Blocking and dilution buffer: 5% NFDM/TBST.
Western blot - Anti-NFkB p105 / p50 antibody [E381] (ab32360)

Anti-NFkB p105 / p50 antibody [E381] (ab32360) at 1/5000 dilution (unpurified) + HeLa cell lysate

Predicted band size: 50 kDa
Observed band size: 105,50 kDa why is the actual band size different from the predicted?
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-NFkB p105 / p50 antibody [E381] (ab32360)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human prostate carcinoma tissue labelling NFkB p105 / p50 with unpurified ab32360 at a dilution of 1/250.
Anti-NFkB p105 / p50 antibody [E381] (ab32360)

IHC-P protocol

Click here to view the general protocols

SDS download

Country/Region

Language
Datasheet download

Download

发表研究结果有使用 ab32360？请让我们知道，以便我们可以引用本数据表中的参考文章。

ab32360 被引用在 210 文献中.

Liu J et al. Tanshinone IIA promotes apoptosis by downregulating BCL2 and upregulating TP53 in triple-negative breast cancer. Naunyn Schmiedebergs Arch Pharmacol 396:365-374 (2023). PubMed: 36374307
Xie J et al. Akkermansia muciniphila protects mice against an emerging tick-borne viral pathogen. Nat Microbiol 8:91-106 (2023). PubMed: 36604506
Wu H et al. Phospholipase D2 targeted by miR-5132-5p alleviates cerulein-induced acute pancreatitis via the Nrf2/NFκB pathway. Immun Inflamm Dis 11:e831 (2023). PubMed: 37249288
Takada H et al. Single-cell transcriptomics uncovers EGFR signaling-mediated gastric progenitor cell differentiation in stomach homeostasis. Nat Commun 14:3750 (2023). PubMed: 37386010
Qi H et al. Knockdown of DEC2 expression inhibits the proliferation of mesangial cells through suppressed glycolysis and p38 MAPK/Erk pathway in lupus nephritis. Mol Med 29:99 (2023). PubMed: 37488524

View all Publications for this product

客户评价及客户问答

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1-10 of 22 Abreviews or Q&A

Application

Immunocytochemistry

Sample

Dog Cell (melanoma cells)

Permeabilization

Yes - Triton X-100

Specification

melanoma cells

Fixative

Formaldehyde

The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Abcam user community

Verified customer

提交于 Apr 08 2022

Application

Western blot

Sample

Dog Cell lysate - whole cell (melanoma ces)

Gel Running Conditions

Reduced Denaturing (12%)

Loading amount

10 µg

Treatment

100 pM canine recombinant IL-1β

Specification

melanoma ces

Blocking step

blockace as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 100%

The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

DR. 令中野

Verified customer

提交于 Apr 08 2022

Application

Western blot

Sample

Human Cell lysate - whole cell (293T)

Gel Running Conditions

Reduced Denaturing (12%)

Loading amount

50000 cells

Specification

293T

Blocking step

Milk as blocking agent for 45 minute(s) · Concentration: 1.5% · Temperature: 21°C

The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Abcam user community

Verified customer

提交于 Aug 14 2019

Application

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)

Sample

Human Tissue sections (Bone marrow)

Antigen retrieval step

Heat mediated - Buffer/Enzyme Used: sodiumcitrate buffer pH 6.0

Permeabilization

Specification

Bone marrow

Blocking step

Optiview Peroxidase Inhibitor as blocking agent for 4 minute(s) · Concentration: 3.0% · Temperature: 36°C

Fixative

Formaldehyde

The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Abcam user community

Verified customer

提交于 Oct 16 2018

Application

Western blot

Sample

Mouse Cell lysate - whole cell (mouse neural stem cell)

Gel Running Conditions

Reduced Denaturing (4-12%)

Loading amount

40 µg

Specification

mouse neural stem cell

Blocking step

BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 20°C

The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Abcam user community

Verified customer

提交于 Jun 26 2018

Application

Immunocytochemistry/ Immunofluorescence

Sample

Human Cell (A375 melanoma cell line)

Permeabilization

Yes - 0,5% Triton-100x

Specification

A375 melanoma cell line

Blocking step

BSA as blocking agent for 30 minute(s) · Concentration: 1% · Temperature: RT°C

Fixative

Paraformaldehyde

The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Cristina Penas Lago

Verified customer

提交于 Oct 05 2017

Application

ChIP

Sample

Rabbit Cell lysate - nuclear (Primary mouse CD4+ Th1 Cells, non activated and ac)

Negative control

Insulin gene (inactive in T cells) (Primer Ins neg).

Specification

Primary mouse CD4+ Th1 Cells, non activated and ac

Detection step

Real-time PCR

Type

Cross-linking (X-ChIP)
Duration of cross-linking step: 10 minute(s) and 0 second(s)
Specification of the cross-linking agent: 1% Formaldehyde

Positive control

Known p50 binding site at the transcriptional start site of NFkB2 (Primer NFkB2_TSS_1) and in the Bcl3 intron (Primer Bcl3 Intr_2).

The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Kathrin Davari

Verified customer

提交于 Sep 28 2016

Application

ChIP

Sample

Mouse Cell lysate - nuclear (Primary CD4+ Th1 cells, non activated or activated)

Negative control

Insulin gene (not active in T cells) (Primer Ins_neg).

Specification

Primary CD4+ Th1 cells, non activated or activated

Detection step

Real-time PCR

Type

Cross-linking (X-ChIP)
Duration of cross-linking step: 10 minute(s) and 0 second(s)
Specification of the cross-linking agent: 1% Formaldehyde

Positive control

Known p50 target site at the transcriptional start site of NFkB2 (Primer: NFkB2_TSS_1).

The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Kathrin Davari

Verified customer

提交于 Sep 27 2016

Application

Western blot

Sample

Rat Tissue lysate - whole (Skeletal muscle)

Gel Running Conditions

Reduced Denaturing (10%)

Loading amount

20 µg

Specification

Skeletal muscle

Blocking step

Milk as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 25°C

The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Abcam user community

Verified customer

提交于 Feb 16 2016

Application

Immunocytochemistry/ Immunofluorescence

Sample

Human Cell (Mueller cell line MIO-M1)

Permeabilization

Yes - 0.1% Tx-100

Specification

Mueller cell line MIO-M1

Blocking step

Serum as blocking agent for 30 minute(s) · Concentration: 5% · Temperature: 25°C

Fixative

Paraformaldehyde

The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Abcam user community

Verified customer

提交于 Dec 31 2015

1-10 of 22 Abreviews or Q&A

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Key features and details

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概述

产品名称

描述

宿主

特异性

经测试应用

种属反应性

免疫原

阳性对照

常规说明

性能

形式

存放说明

存储溶液

纯度

克隆

克隆编号

同种型

研究领域

相关产品

Alternative Versions

Assay kits

Compatible Secondaries

Isotype control

Positive Controls

应用

The Abpromise guarantee

靶标

功能

序列相似性

结构域

翻译后修饰

细胞定位

数据库链接

别名

图片

实验方案

数据表及文件

SDS download

Datasheet download

文献 (210)

客户评价及客户问答

Immunocytochemistry abreview for Anti-NFkB p105 / p50 antibody [E381]

Western blot abreview for Anti-NFkB p105 / p50 antibody [E381]

Western blot abreview for Anti-NFkB p105 / p50 antibody [E381]

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-NFkB p105 / p50 antibody [E381]

Western blot abreview for Anti-NFkB p105 / p50 antibody [E381]

Immunocytochemistry/ Immunofluorescence abreview for Anti-NFkB p105 / p50 antibody [E381]

ChIP abreview for Anti-NFkB p105 / p50 antibody [E381]

ChIP abreview for Anti-NFkB p105 / p50 antibody [E381]

Western blot abreview for Anti-NFkB p105 / p50 antibody [E381]

Immunocytochemistry/ Immunofluorescence abreview for Anti-NFkB p105 / p50 antibody [E381]