Recombinant Anti-Bmi1 antibody [EPR3745(2)] (ab126783)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR3745(2)] to Bmi1 - ChIP Grade
- Suitable for: IP, ChIC/CUT&RUN-seq, WB, IHC-P, ICC/IF
- Knockout validated
- Reacts with: Rat, Human
Related conjugates and formulations
Overview
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Product name
Anti-Bmi1 antibody [EPR3745(2)]
See all Bmi1 primary antibodies -
Description
Rabbit monoclonal [EPR3745(2)] to Bmi1 - ChIP Grade -
Host species
Rabbit -
Tested applications
Suitable for: IP, ChIC/CUT&RUN-seq, WB, IHC-P, ICC/IFmore details -
Species reactivity
Reacts with: Rat, Human -
Immunogen
Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
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Positive control
- WB: MCF7, A431, HEK293T, K562, SAOS-2, SW480, MOLT4, PC-12 and HT1080 cell lysates. IHC-P: Human tonsil, colonic adenocarcinoma, lung adenocarcinoma, breast carcinoma and thyroid gland carcinoma tissues. ICC/IF: SW480 and HeLa cells. IP: K-562 cell lysate ChIP/CUT&RUN Seq: NCCIT
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General notes
Mouse: Internal data indicated that the antibody is not suitable for WB application in mouse species.
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
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated 'PUR' on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.
Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C. Stable for 12 months at -20°C. -
Storage buffer
pH: 7.20
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol, 0.05% BSA -
Concentration information loading...
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Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR3745(2) -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
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Compatible Secondaries
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Isotype control
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KO cell lines
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KO cell lysates
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KO cell pellets
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Positive Controls
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Recombinant Protein
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab126783 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Application | Abreviews | Notes |
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IP |
1/50.
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ChIC/CUT&RUN-seq |
Use at an assay dependent concentration.
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WB | (2) |
1/10000 - 1/50000. Detects a band of approximately 40 kDa (predicted molecular weight: 36 kDa).
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IHC-P |
1/100 - 1/500. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
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ICC/IF |
1/100 - 1/500.
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Notes |
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IP
1/50. |
ChIC/CUT&RUN-seq
Use at an assay dependent concentration. |
WB
1/10000 - 1/50000. Detects a band of approximately 40 kDa (predicted molecular weight: 36 kDa). |
IHC-P
1/100 - 1/500. Perform heat mediated antigen retrieval before commencing with IHC staining protocol. |
ICC/IF
1/100 - 1/500. |
Target
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Function
Component of the Polycomb group (PcG) multiprotein PRC1 complex, a complex required to maintain the transcriptionally repressive state of many genes, including Hox genes, throughout development. PcG PRC1 complex acts via chromatin remodeling and modification of histones; it mediates monoubiquitination of histone H2A 'Lys-119', rendering chromatin heritably changed in its expressibility. In the PRC1 complex, it is required to stimulate the E3 ubiquitin-protein ligase activity of RNF2/RING2. -
Sequence similarities
Contains 1 RING-type zinc finger. -
Post-translational
modificationsMonoubiquitinated (By similarity). May be polyubiquitinated; which does not lead to proteasomal degradation. -
Cellular localization
Nucleus. Cytoplasm. - Information by UniProt
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Database links
- Entrez Gene: 100532731 Human
- Entrez Gene: 648 Human
- Entrez Gene: 307151 Rat
- Omim: 164831 Human
- SwissProt: P35226 Human
- Unigene: 380403 Human
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Alternative names
- B lymphoma Mo MLV insertion region (mouse) antibody
- B lymphoma Mo MLV insertion region 1 homolog antibody
- Bmi 1 antibody
see all
Images
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All lanes : Anti-Bmi1 antibody [EPR3745(2)] (ab126783) at 1/10000 dilution
Lane 1 : Wild-type MCF7 cell lysate
Lane 2 : BMI1 knockout MCF7 cell lysate
Lane 3 : A431 cell lysate
Lysates/proteins at 20 µg per lane.
Performed under reducing conditions.
Predicted band size: 36 kDa
Observed band size: 37 kDa why is the actual band size different from the predicted?Lanes 1- 3: Merged signal (red and green). Green - ab126783 observed at 37 kDa. Red - Anti-alpha Tubulin antibody [DM1A] - Loading Control (ab7291) observed at 50 kDa.
ab126783 was shown to react with Bmi1 in wild-type MCF7 cells in western blot. Loss of signal was observed when knockout cell line ab262319 (knockout cell lysate ab256851) was used. Wild-type MCF7 and BMI1 knockout MCF7 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. ab126783 and Anti-alpha Tubulin antibody [DM1A] - Loading Control (ab7291) overnight at 4°C at a 1 in 10000 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.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Bmi1 antibody [EPR3745(2)] (ab126783)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human breast carcinoma tissue labelling Bmi1 with purified ab126783 at 1/500. 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.
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Immunocytochemistry/Immunofluorescence analysis of HeLa cells labelling Bmi1 with purified ab126783 at 1/500. Cells were fixed with 4% paraformaldehyde and permeabilized with 0.1% Triton X-100. ab150077, an Alexa Fluor® 488-conjugated goat anti-rabbit IgG (1/500) was used as the secondary antibody. DAPI (blue) was used as the nuclear counterstain. ab7291, a mouse anti-tubulin (1/1000) and ab150120, an Alexa Fluor® 594-conjugated goat anti-mouse IgG (1/500) were also used.
Control 1: primary antibody (1/500) and secondary antibody, ab150120, an Alexa Fluor® 594-conjugated goat anti-mouse IgG (1/500).
Control 2: ab7291 (1/1000) and secondary antibody, ab150077, an Alexa Fluor® 488-conjugated goat anti-rabbit IgG (1/500).
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Lane 1: Wild-type HAP1 cell lysate (20 µg)
Lane 2: Bmi1 knockout HAP1 cell lysate (20 µg)
Lane 3: U2OS cell lysate (20 µg)
Lane 4: Molt-4 cell lysate (20 µg)
Lanes 1 - 4: Merged signal (red and green). Green - ab126783 observed at 42 kDa. Red - loading control, ab18058, observed at 37 kDa.ab126783 was shown to specifically react with Bmi1 when Bmi1 knockout samples were used. Wild-type and Bmi1 knockout samples were subjected to SDS-PAGE. ab126783 and ab18058 (loading control to Vinculin) were both diluted at 1/10 000 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 1/10 000 dilution for 1 h at room temperature before imaging. -
All lanes : Anti-Bmi1 antibody [EPR3745(2)] (ab126783) at 1/1000 dilution
Lane 1 : Wild-type HEK293T cell lysate
Lane 2 : BMI1 knockout HEK293T cell lysate
Lane 3 : MOLT-4 cell lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) at 1/10000 dilution
Predicted band size: 36 kDa
Observed band size: 37 kDa why is the actual band size different from the predicted?Lanes 1-3: Merged signal (red and green). Green - ab126783 observed at 37 kDa. Red - loading control ab7291 observed at 50 kDa.
ab126783 Anti-Bmi1 antibody [EPR3745(2)] was shown to specifically react with Bmi1 in wild-type HEK293T cells. Loss of signal was observed when knockout cell line ab266514 (knockout cell lysate ab256850) was used. Wild-type and Bmi1 knockout samples were subjected to SDS-PAGE. ab126783 and Anti-alpha Tubulin antibody [DM1A] - Loading Control (ab7291) were incubated at room temperature for 2. 5 hours at 1 in 1000 dilution and 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.
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ab126783 (purified) at 1/50 immunoprecipitating Bmi1 in 10 μg K-562 (Human chronic myelogenous leukemia lymphoblast) whole cell lysate (Lanes 1 and 2, observed at 43 kDa). Lane 3 - Rabbit monoclonal IgG (ab172730) instead of ab126783 in K-562 whole cell lysate. For western blotting, ab126783 at 1/500 and VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/1000 dilution.
Blocking/Dilution buffer and concentration: 5% NFDM/TBST.
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Anti-Bmi1 antibody [EPR3745(2)] (ab126783) at 1/20000 dilution (purified) + PC-12 cell lysate at 20 µg
Secondary
Peroxidase-conjugated goat anti-rabbit IgG, (H+L) at 1/1000 dilution
Predicted band size: 36 kDa
Observed band size: 40 kDa why is the actual band size different from the predicted?Blocking and dilution buffer: 5% NFDM/TBST.
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All lanes : Anti-Bmi1 antibody [EPR3745(2)] (ab126783) at 1/20000 dilution (purified)
Lane 1 : K562 cell lysate
Lane 2 : SAOS-2 cell lysate
Lane 3 : SW480 cell lysate
Lane 4 : Molt-4 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: 36 kDa
Observed band size: 40 kDa why is the actual band size different from the predicted?Blocking and dilution buffer: 5% NFDM/TBST.
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All lanes : Anti-Bmi1 antibody [EPR3745(2)] (ab126783) at 1/10000 dilution (unpurified)
Lane 1 : K562 cell lysate
Lane 2 : SAOS-2 cell lysate
Lane 3 : SW480 cell lysate
Lane 4 : MOLT4 cell lysate
Lane 5 : HT1080 cell lysate
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : HRP-conjugated goat anti-rabbit IgG at 1/2000 dilution
Predicted band size: 36 kDa -
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Bmi1 antibody [EPR3745(2)] (ab126783)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human normal tonsil tissue labelling Bmi1 with unpurifiied ab126783.
Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Bmi1 antibody [EPR3745(2)] (ab126783)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human colonic adenocarcinoma tissue labelling Bmi1 with unpurifiied ab126783.
Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Bmi1 antibody [EPR3745(2)] (ab126783)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human lung adenocarcinoma tissue labelling Bmi1 with unpurifiied ab126783.
Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Bmi1 antibody [EPR3745(2)] (ab126783)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human thyroid gland carcinoma tissue labelling Bmi1 with unpurifiied ab126783.
Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
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Immunocytochemistry/Immunofluorescence analysis of SW480 cells labelling Bmi1 with unpurified ab126783 at a dilution of 1/100.
Protocols
Datasheets and documents
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SDS download
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Datasheet download
References (26)
ab126783 has been referenced in 26 publications.
- Wu Z et al. The inhibitory effect of human DEFA5 in growth of gastric cancer by targeting BMI1. Cancer Sci 112:1075-1083 (2021). PubMed: 33503272
- Li S et al. Schisandrin B inhibits epithelial-mesenchymal transition and stemness of large-cell lung cancer cells and tumorigenesis in xenografts via inhibiting the NF-?B and p38 MAPK signaling pathways. Oncol Rep 45:N/A (2021). PubMed: 33907830
- Chen Y et al. Effects and mechanism of microRNA-218 against lung cancer. Mol Med Rep 23:N/A (2021). PubMed: 33179084
- Teng Y et al. Cytoplasmic p27 is a novel prognostic biomarker and oncogenic protein for nasopharyngeal carcinoma. Artif Cells Nanomed Biotechnol 48:336-344 (2020). PubMed: 31884829
- Luo X et al. miR-96-5p Suppresses the Progression of Nasopharyngeal Carcinoma by Targeting CDK1. Onco Targets Ther 13:7467-7477 (2020). PubMed: 32801769