BMI-1 antibody (mAb)

RRID: AB_2615070
Clone: AF27
Aliases: PCGF4, RNF51, FLVI2/BMI1
Catalog No: 39993 Format: 100 µg $435 Buy
Catalog No: 39994 Format: 10 µg $105 Buy

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Antibody Type:
Monoclonal
Isotype:
IgG2a
Purification:
Protein G Chromatography
Host:
Mouse
Molecular Weight:
40 kDa
Reactivity:
Human, Mouse

Applications

ChIP Validated ChIP-Seq Validated Western Blot Validated

Application Notes


Validated Applications:
ChIP-Seq: 4 µg per ChIP
WB: 0.5 - 2 µg/mL

For optimal results, we recommend the addition of 0.05% Tween 20 to all blocking solutions to reduce background. Individual optimization may be required.

Published Applications:
ChIP
WB
IHC (FFPE)
IF


See references for more information.

Also available as an AbFlex® engineered recombinant antibody. For details on the corresponding AbFlex Recombinant Antibody, see Catalog No. 91195.

Immunogen

This BMI-1 antibody was raised against a recombinant protein corresponding to full-length mouse BMI-1.

Buffer

Purified IgG in 70 mM Tris (pH 8), 105 mM NaCl, 31 mM glycine, 0.07 mM EDTA, 30% glycerol and 0.035% sodium azide. Sodium azide is highly toxic.

References

 

BMI-1 antibody (mAb) tested by ChIP-Seq.
Chromatin immunoprecipitation (ChIP) was performed using the ChIP-IT® High Sensitivity Kit (Cat. No. 53040) with 30 µg of chromatin from mouse embryonic fibroblast (MEF) cells and 4 µg BMI-1 antibody. ChIP DNA was sequenced on the Illumina NextSeq and 10 million sequence tags were mapped to identify BMI-1 binding sites.

BMI-1 antibody (mAb) tested by Western blot.
20 µg of K562 cell nuclear extract was run on SDS-PAGE and probed with BMI-1 antibody at 0.5 µg/ml. For optimal results, we recommend the addition of 0.05% Tween 20 to all blocking solutions to reduce background.

Background

BMI-1 is a member of the Polycomb PRC1 complex that is recruited to transcriptionally repressed genes subsequent to histone H3 lysine 27 methylation in order to maintain repression. In the PRC1 complex, BMI-1 is required to stimulate the E3 ubiquitin-protein ligase activity of RNF2/RING2, resulting in the ubiquitylation of histone H2A at lysine 119. BMI-1 acts as an oncogene and cooperates with c-myc in the initiation of lymphoma. High levels of expression of BMI-1 are observed in metastatic melanoma, and BMI-1 has been implicated in several other cancers. BMI-1 is required for hematopoietic stem cell self renewal.

Learn more about the PRC1 Complex and Stem Cell Identity

Chromatin IP Assays

This antibody has been validated for use in ChIP and/or ChIP-Seq, and can be used with Active Motif's ChIP-IT® High Sensitivity Kit or our magnetic bead-based ChIP-IT® Express Kits. For an overview of all of our ChIP products, please visit our Chromatin IP Page to learn about ChIP products designed for use with BMI-1 antibody (mAb).

Bridging Antibody to Improve ChIP, MeDIP and IP

Some isotypes of mouse IgG antibodies do not bind well to protein G-conjugated agarose/magnetic beads, which are commonly used in ChIP, MeDIP and IP experiments. Capture efficiency can be improved by including an anti-mouse IgG bridging antibody because it binds with a strong affinity to both the protein G beads and the mouse IgG antibody. By increasing the affinity of the mouse primary antibody for the protein G bead, the Bridging Antibody for Mouse IgG can help improve the results of all ChIP, MeDIP and IP experiments that use BMI-1 antibody (mAb).

Storage

Some products may be shipped at room temperature. This will not affect their stability or performance. Avoid repeated freeze/thaw cycles by aliquoting items into single-use fractions for storage at -20°C for up to 2 years. Keep all reagents on ice when not in storage.

Guarantee

This product is guaranteed for 12 months from date of receipt.

This product is for research use only and is not for use in diagnostic procedures.

Application Key

  • ChIP = Chromatin Immunoprecipitation;
  • DB = Dot Blot;
  • ELISA = Enzyme-linked Immunosorbent Assay;
  • EMSA = Electrophoretic Mobility Shift Assay
  • FC = Flow Cytometry;
  • ICC = Immunocytochemistry;
  • IF = Immunofluorescence;
  • IHC = Immunohistochemistry;
  • IP = Immunoprecipitation;
  • MeDIP = Methyl-DNA Immunoprecipitation;
  • TR-FRET = Time-Resolved Fluorescence Resonance Energy Transfer;
  • WB = Western Blotting