AbFlex® BMI-1 antibody (rAb)

RRID: AB_2793798
Catalog No: 91195 Format: 100 µg $385 Buy
Catalog No: 91196 Format: 10 µg $80 Buy

Request a quote for a bulk order Bulk Order Button


Antibody Type:
Recombinant
Isotype:
IgG2a
Purification:
Protein A Chromatography
Host:
Mouse
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 dilution.


AbFlex™ recombinant antibodies are genetically derived from DNA sequences of parental hybridoma clones.  For details on the parental clone, see Catalog No. 39993.

Note: many chromatin-bound proteins are not soluble in a low salt nuclear extract and fractionate to the pellet. Therefore, we recommend a High Salt / Sonication Protocol when preparing nuclear extracts for Western Blot.

Immunogen

This antibody was raised against full length human BMI1 [BMI1 polycomb ring finger oncogene].

Buffer

Purified IgG in 140 mM Hepes, pH 7.5, 70 mM NaCl, 35 mM NaOAc, 0.07% sodium azide, 30% glycerol.

References

 

Western blot of AbFlex® BMI-1 antibody (rAb).
20 µg of K562 cell nuclear extract was run on SDS-PAGE and probed with AbFlex® BMI-1 antibody.

AbFlex® BMI-1 recombinant antibody (rAb) tested by ChIP-Seq
Chromatin immunoprecipitation (ChIP) was performed using the ChIP-IT® High Sensitivity Kit (Cat. No. 53040) with 30 µg of mouse embryonic fibroblast (MEF) chromatin and 4 µg of BMI-1 antibody. ChIP DNA was sequenced on the Illumina NextSeq and 9.9 million sequence tags were mapped to identify BMI-1 binding sites on chromosome 2.

Background

AbFlex® antibodies are recombinant antibodies (rAbs) that have been generated using defined DNA sequences to produce highly specific, reproducible antibodies. Each AbFlex antibody contains a 6xHis-Tag, an avidin tag sequence for enzymatic biotin conjugation using the biotin ligase, BirA, and a sortase recognition motif (LPXTG) to attach a variety of labels directly to the antibody including fluorophores, enzymatic substrates (HRP, AP), peptides, drugs as well as solid supports.  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 AbFlex® BMI-1 antibody (rAb).

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 AbFlex® BMI-1 antibody (rAb).

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