Name: Anti-Bmi1 antibody [EPR3745(2)]
Brand: RabMAb
SKU: ab126783
Rating: 3 (3 reviews)

Product name

Anti-Bmi1 antibody [EPR3745(2)]
See all Bmi1 primary antibodies
Description

Rabbit monoclonal [EPR3745(2)] to Bmi1
Host species

Rabbit
Tested applications

Suitable for: IP, WB, IHC-P, ICC/IFmore details
Species reactivity

Reacts with: Rat, Human
Immunogen

A synthetic peptide, corresponding to residues in Human Bmi1 (UniProt P35226).
Positive control
- WB: 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
General notes

Mouse: Internal data indicated that the antibody is not suitable for WB application in mouse species.
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.
This product is a recombinant rabbit monoclonal antibody.

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...
Purity

Protein A purified
Clonality

Monoclonal
Clone number

EPR3745(2)
Isotype

IgG
Research areas

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
IP		1/500.
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. See IHC antigen retrieval protocols.
ICC/IF		1/100 - 1/500.

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
modifications

Monoubiquitinated (By similarity). May be polyubiquitinated; which does not lead to proteasomal degradation.
Cellular localization

Nucleus. Cytoplasm.
Target information above from: UniProt accession P35226 The UniProt Consortium
The Universal Protein Resource (UniProt) in 2010
Nucleic Acids Res. 38:D142-D148 (2010) .

Information by UniProt
Database links
- Entrez Gene: 100532731 Human
- Entrez Gene: 648 Human
- Entrez Gene: 307151 Rat
- Omim: 164831 Human
- SwissProt: P35226 Human
- Unigene: 380403 Human
Alternative names
- B lymphoma Mo MLV insertion region (mouse) antibody
- B lymphoma Mo MLV insertion region 1 homolog antibody
- Bmi 1 antibody
- BMI1 antibody
- BMI1 polycomb ring finger oncogene antibody
- BMI1_HUMAN antibody
- Flvi 2/bmi 1 antibody
- FLVI2/BMI1 antibody
- MGC12685 antibody
- Murine leukemia viral (bmi 1) oncogene homolog antibody
- Oncogene BMI 1 antibody
- PCGF 4 antibody
- PCGF4 antibody
- Polycomb complex protein BMI 1 antibody
- Polycomb complex protein BMI-1 antibody
- Polycomb group protein Bmi1 antibody
- Polycomb group ring finger 4 antibody
- Polycomb group RING finger protein 4 antibody
- RING finger protein 51 antibody
- RNF 51 antibody
- RNF51 antibody
see all

Western blot - Anti-Bmi1 antibody [EPR3745(2)] (ab126783)

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.
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.
Immunocytochemistry/ Immunofluorescence - Anti-Bmi1 antibody [EPR3745(2)] (ab126783)

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).
Immunoprecipitation - Anti-Bmi1 antibody [EPR3745(2)] (ab126783)

ab126783 (purified) at 1/500 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, HRP Veriblot for IP (ab131366) was used as the secondary antibody (1/1000).

Blocking/Dilution buffer and concentration: 5% NFDM/TBST.
Western blot - Anti-Bmi1 antibody [EPR3745(2)] (ab126783)

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.
Western blot - Anti-Bmi1 antibody [EPR3745(2)] (ab126783)

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.
Western blot - Anti-Bmi1 antibody [EPR3745(2)] (ab126783)

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.
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.
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.
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.
Immunocytochemistry/ Immunofluorescence - Anti-Bmi1 antibody [EPR3745(2)] (ab126783)

Immunocytochemistry/Immunofluorescence analysis of SW480 cells labelling Bmi1 with unpurified ab126783 at a dilution of 1/100.

Western blot protocols
Immunohistochemistry protocols
Immunocytochemistry & immunofluorescence protocols

This product has been referenced in:

Kaowinn S et al. STAT1-HDAC4 signaling induces epithelial-mesenchymal transition and sphere formation of cancer cells overexpressing the oncogene, CUG2. Oncol Rep 40:2619-2627 (2018). Read more (PubMed: 30226605) »
Ji D et al. Enhancement of Sensitivity to Chemo/Radiation Therapy by Using miR-15b against DCLK1 in Colorectal Cancer. Stem Cell Reports 11:1506-1522 (2018). Read more (PubMed: 30449704) »

See all 13 Publications for this product

Publishing research using ab126783? Please let us know so that we can cite the reference in this datasheet.

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1-3 of 3 Abreviews

Application

Western blot

Sample

Mouse Tissue lysate - whole (Prefrontal cortex and hippocampus)

Gel Running Conditions

Reduced Denaturing (3-8% Tris-Acetate)

Loading amount

20 µg

Specification

Prefrontal cortex and hippocampus

Blocking step

Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 23°C

Abcam user community

Verified customer

Submitted Jan 04 2018

Application

Western blot

Loading amount

100 µg

Gel Running Conditions

Reduced Denaturing (4-20)

Sample

Mouse Cell lysate - nuclear (neural stem cells)

Specification

neural stem cells

Blocking step

licor PBS blocking buffer as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 100% · Temperature: 20°C

Abcam user community

Verified customer

Submitted Dec 11 2014

Application

Immunocytochemistry/ Immunofluorescence

Blocking step

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

Sample

Mouse Cell (heart)

Specification

heart

Permeabilization

Fixative

Acetone

Abcam user community

Verified customer

Submitted Dec 10 2014

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

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Anti-Bmi1 antibody [EPR3745(2)] (ab126783)

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