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AbFlex® BMI-1 antibody (rAb)

Catalog No: 91195 Format: 100 µg ¥62,000 Buy Now
Catalog No: 91196 Format: 10 µg ¥17,000 Buy Now

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Antibody Type:Recombinant
Isotype:IgG2a
Purification:Protein A Chromatography
Host:Mouse
Reactivity:Human, Mouse

Applications

ChIP Validated ChIP-Seq Validated Western Blot Validated

for AbFlex BMI-1 antibody (rAb)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.

for AbFlex BMI-1 antibody (rAb)Immunogen

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

for AbFlex BMI-1 antibody (rAb)Buffer

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

for AbFlex BMI-1 antibody (rAb)References

 
AbFlex<sup>®</sup> BMI-1 antibody (rAb) tested by Western blot.

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

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.

for AbFlex BMI-1 antibody (rAb)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).

for AbFlex BMI-1 antibody (rAb)Storage

Some products may be shipped at room temperature. This will not affect their stability or performance. Upon receipt, unconjugated antibodies may be stored at -20°C for up to 2 years. Fluorophore- & enzyme-conjugated antibodies should be stored at 4°C. Fluorophore-conjugated antibodies should be protected from light. Keep reagents on ice when not in storage; to avoid repeated freeze/thaw cycles, we recommend aliquoting items that will be stored frozen into single-use fractions prior to freezing.

for AbFlex BMI-1 antibody (rAb)Guarantee

This product is guaranteed for 6 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
 

Technical Data Sheet

AbFlex® BMI-1 antibody (rAb)
 

Data Thumbnails

AbFlex<sup>®</sup> BMI-1 antibody (rAb) tested by Western blot.

Western blot of AbFlex® BMI-1 antibody (rAb).
(Click image to enlarge and see details.)

AbFlex® BMI-1 recombinant antibody (rAb) tested by ChIP-Seq

AbFlex® BMI-1 recombinant antibody (rAb) tested by ChIP-Seq
(Click to enlarge and view details)