Description
Product Usage Information
For optimal ChIP and ChIP-seq results, use 10 μl of antibody and 10 μg of chromatin (approximately 4 x 10 6 cells) per IP. This antibody has been validated using SimpleChIP ® Enzymatic Chromatin IP Kits. The CUT&RUN dilution was determined using CUT&RUN Assay Kit # 86652 . The CUT&Tag dilution was determined using CUT&Tag Assay Kit # 77552 .
Western Blotting: 1:1000
Immunoprecipitation: 1:200
Immunohistochemistry (Paraffin): 1:200
Immunofluorescence (Immunocytochemistry): 1:600
Flow Cytometry (Fixed/Permeabilized): 1:100
Chromatin IP: 1:50
Chromatin IP-seq: 1:50
CUT&RUN: 1:50
CUT&Tag: 1:50
Storage
Supplied in 10 mM sodium HEPES (pH 7.5), 150 mM NaCl, 100 µg/ml BSA, 50% glycerol and less than 0.02% sodium azide. Store at -20°C. Do not aliquot the antibody. For a carrier free (BSA and azide free) version of this product see product # 39168 .
Protocol
Available protocols: Western Blotting, Immunoprecipitation, Immunohistochemistry (Paraffin), Immunofluorescence (Immunocytochemistry), Flow Cytometry (Fixed/Permeabilized), Chromatin IP, Chromatin IP-seq, CUT&RUN, CUT&Tag
Specificity / Sensitivity
Bmi1 (D20B7) Rabbit Monoclonal Antibody recognizes endogenous levels of total Bmi1 protein.
Species Reactivity: Human, Monkey
Source / Purification
Monoclonal antibody is produced by immunizing animals with recombinant protein specific to the carboxy terminus of human Bmi1 protein.
Background
The polycomb group (PcG) of proteins contributes to the maintenance of cell identity, stem cell self-renewal, cell cycle regulation, and oncogenesis by maintaining the silenced state of genes that promote cell lineage specification, cell death, and cell-cycle arrest (1-4). PcG proteins exist in two complexes that cooperate to maintain long-term gene silencing through epigenetic chromatin modifications. The first complex, EED-EZH2, is recruited to genes by DNA-binding transcription factors and methylates histone H3 on Lys27. This histone methyl-transferase activity requires the Ezh2, Eed, and Suz12 subunits of the complex (5). Histone H3 methylation at Lys27 facilitates the recruitment of the second complex, PRC1, which ubiquitinylates histone H2A on Lys119 (6). Bmi1 is a component of the PRC1 complex, which together with Ring1 strongly enhances the E3 ubiquitin ligase activity of the Ring2 catalytic subunit (7). Bmi1 plays an important role in the regulation of cell proliferation and senescence through repression of the p16 INK4A and p19 ARF genes and is required for maintenance of adult hematopoietic and neural stem cells (3,4,8-10).
Alternate Names
B lymphoma Mo-MLV insertion region 1 homolog; BMI1; BMI1 polycomb ring finger oncogene; BMI1 polycomb ring finger proto-oncogene; BMI1 proto-oncogene, polycomb ring finger; flvi-2/bmi-1; FLVI2/BMI1; MGC12685; murine leukemia viral (bmi-1) oncogene homolog; oncogene BMI-1; PCGF4; Polycomb complex protein BMI-1; polycomb group protein Bmi1; polycomb group ring finger 4; Polycomb group RING finger protein 4; RING finger protein 51; RNF51
