Revision 6

#5856

Store at -20C

Bmi1 (D42B3) Rabbit mAb
CST Logo
Orders:

877-616-CELL (2355)

[email protected]

Support:

877-678-TECH (8324)

Web:

3 Trask Lane | Danvers | Massachusetts | 01923 | USA

For Research Use Only. Not for Use in Diagnostic Procedures.

Applications:
W, IP, IF-IC, ChIP, ChIP-seq, C&R, C&T

Reactivity:
H M R Mk

Sensitivity:
Endogenous

MW (kDa):
41, 43

Source/Isotype:
Rabbit IgG

UniProt ID:
#P35226

Entrez-Gene Id:
648

Product Usage Information

For optimal ChIP and ChIP-seq results, use 10 μl of antibody and 10 μg of chromatin (approximately 4 x 106 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.

Application Dilution
Western Blotting 1:1000
Immunoprecipitation 1:100
Immunofluorescence (Immunocytochemistry) 1:50 - 1:200
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.

Specificity/Sensitivity

Bmi1 (D42B3) Rabbit mAb recognizes endogenous levels of total Bmi1 protein.

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).

Background References

Species Reactivity

Species reactivity is determined by testing in at least one approved application (e.g., western blot).

Western Blot Buffer

IMPORTANT: For western blots, incubate membrane with diluted primary antibody in 5% w/v BSA, 1X TBS, 0.1% Tween® 20 at 4°C with gentle shaking, overnight.

Applications Key

W: Western Blotting IP: Immunoprecipitation IF-IC: Immunofluorescence (Immunocytochemistry) ChIP: Chromatin IP ChIP-seq: Chromatin IP-seq C&R: CUT&RUN C&T: CUT&Tag

Cross-Reactivity Key

H: Human M: Mouse R: Rat Mk: Monkey

Trademarks and Patents

Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.

All other trademarks are the property of their respective owners. Visit cellsignal.com/trademarks for more information.

限制使用

除非 CST 的合法授书代表以书面形式书行明确同意，否书以下条款适用于 CST、其关书方或分书商提供的书品。 任何书充本条款或与本条款不同的客书条款和条件，除非书 CST 的合法授书代表以书面形式书独接受， 否书均被拒书，并且无效。

专品专有“专供研究使用”的专专或专似的专专声明， 且未专得美国食品和专品管理局或其他外国或国内专管机专专专任何用途的批准、准专或专可。客专不得将任何专品用于任何专断或治专目的， 或以任何不符合专专声明的方式使用专品。CST 专售或专可的专品提供专作专最专用专的客专，且专用于研专用途。将专品用于专断、专防或治专目的， 或专专售（专独或作专专成）或其他商专目的而专专专品，均需要 CST 的专独专可。客专：(a) 不得专独或与其他材料专合向任何第三方出售、专可、 出借、捐专或以其他方式专专或提供任何专品，或使用专品制造任何商专专品，(b) 不得复制、修改、逆向工程、反专专、 反专专专品或以其他方式专专专专专品的基专专专或技专，或使用专品开专任何与 CST 的专品或服专专争的专品或服专， (c) 不得更改或专除专品上的任何商专、商品名称、徽专、专利或版专声明或专专，(d) 只能根据 CST 的专品专售条款和任何适用文档使用专品 ， (e) 专遵守客专与专品一起使用的任何第三方专品或服专的任何专可、服专条款或专似专专

Orders: 877-616-CELL (2355) • [email protected] • Support: 877-678-TECH (8324) • [email protected] • Web: cellsignal.com
For Research Use Only. Not for Use in Diagnostic Procedures.

Revision 6

#5856

Bmi1 (D42B3) Rabbit mAb

Cell Signaling Technology Logo
Western blot analysis of extracts from various cell lines using Bmi1 (D42B3) Rabbit mAb.
Western Blotting Image 1: Bmi1 (D42B3) Rabbit mAb
Confocal immunofluorescent analysis of C2C12 cells using Bmi1 (D42B3) Rabbit mAb (green). Actin filaments were labeled with DY-554 phalloidin (red).
Immunofluorescence Image 1: Bmi1 (D42B3) Rabbit mAb
Chromatin immunoprecipitations were performed with cross-linked chromatin from NCCIT cells and either Bmi1 (D42B3) Rabbit mAb or RING1B (D22F2) XP® Rabbit mAb #5694, using SimpleChIP® Enzymatic Chromatin IP Kit (Magnetic Beads) #9005. DNA Libraries were prepared using DNA Library Prep Kit for Illumina® (ChIP-seq, CUT&RUN) #56795. Bmi1 and RING1B are known to associate with each other on chromatin. The figure shows binding of both Bmi1 and RING1B across HOXD gene cluster, which are known target genes of Bmi1 and RING1B (see additional figure containing ChIP-qPCR data). For additional ChIP-seq tracks, please download the product data sheet.
Chromatin Immunoprecipitation Image 1: Bmi1 (D42B3) Rabbit mAb
Orders: 877-616-CELL (2355) • [email protected] • Support: 877-678-TECH (8324) • [email protected] • Web: cellsignal.com
For Research Use Only. Not for Use in Diagnostic Procedures.

Revision 6

#5856

Bmi1 (D42B3) Rabbit mAb

Cell Signaling Technology Logo
Chromatin immunoprecipitations were performed with cross-linked chromatin from NCCIT cells and either Bmi1 (D42B3) Rabbit mAb or RING1B (D22F2) XP® Rabbit mAb #5694, using SimpleChIP® Enzymatic Chromatin IP Kit (Magnetic Beads) #9005. DNA Libraries were prepared using DNA Library Prep Kit for Illumina® (ChIP-seq, CUT&RUN) #56795. Bmi1 and RING1B are known to associate with each other on chromatin. The figure shows binding of both Bmi1 and RING1B across HOXA gene cluster (upper) and HOXD gene cluster (lower), which are known target genes of Bmi1 and RING1B (see additional figure containing ChIP-qPCR data).
Chromatin Immunoprecipitation Image 2: Bmi1 (D42B3) Rabbit mAb
Chromatin immunoprecipitations were performed with cross-linked chromatin from NCCIT cells and either Bmi1 (D42B3) Rabbit mAb or Normal Rabbit IgG #2729 using SimpleChIP® Enzymatic Chromatin IP Kit (Magnetic Beads) #9003. The enriched DNA was quantified by real-time PCR using SimpleChIP® Human HoxA1 Intron 1 Primers #7707, SimpleChIP® Human HoxA2 Promoter Primers #5517, and SimpleChIP® Human α Satellite Repeat Primers #4486. The amount of immunoprecipitated DNA in each sample is represented as signal relative to the total amount of input chromatin (equivalent to one).
Chromatin Immunoprecipitation Image 3: Bmi1 (D42B3) Rabbit mAb
CUT&RUN was performed with NCCIT cells and either Bmi1 (D42B3) Rabbit mAb or RING1B (D22F2) XP® Rabbit mAb #5694, using CUT&RUN Assay Kit #86652. DNA Libraries were prepared using DNA Library Prep Kit for Illumina® (ChIP-seq, CUT&RUN) #56795. RING1B and Bmi1 are both PRC1 components. The figure shows binding across COMMD3 gene.
CUT & RUN Image 1: Bmi1 (D42B3) Rabbit mAb
Orders: 877-616-CELL (2355) • [email protected] • Support: 877-678-TECH (8324) • [email protected] • Web: cellsignal.com
For Research Use Only. Not for Use in Diagnostic Procedures.

Revision 6

#5856

Bmi1 (D42B3) Rabbit mAb

Cell Signaling Technology Logo
CUT&RUN was performed with NCCIT cells and either Bmi1 (D42B3) Rabbit mAb or RING1B (D22F2) XP® Rabbit mAb #5694, using CUT&RUN Assay Kit #86652. DNA Libraries were prepared using DNA Library Prep Kit for Illumina® (ChIP-seq, CUT&RUN) #56795. RING1B and Bmi1 are both PRC1 components. The figures show binding across HOXD (upper) and COMMD3 (lower) genes.
CUT & RUN Image 2: Bmi1 (D42B3) Rabbit mAb
CUT&RUN was performed with NCCIT cells and either Bmi1 (D42B3) Rabbit mAb or Rabbit (DA1E) mAb IgG XP® Isotype Control (CUT&RUN) #66362, using CUT&RUN Assay Kit #86652. The enriched DNA was quantified by real-time PCR using SimpleChIP® Human HoxA1 Intron 1 Primers #7707, SimpleChIP® Human HoxA2 Promoter Primers #5517, and SimpleChIP® Human α Satellite Repeat Primers #4486. The amount of immunoprecipitated DNA in each sample is represented as signal relative to the total amount of input chromatin, which is equivalent to one.
CUT & RUN Image 3: Bmi1 (D42B3) Rabbit mAb
CUT&Tag was performed with NCCIT cells and Bmi1 (D42B3) Rabbit mAb, using CUT&Tag Assay Kit #77552. DNA library was prepared using CUT&Tag Dual Index Primers and PCR Master Mix for Illumina Systems #47415. The figure shows binding across HOXA1, a known target gene of Bmi1 (see our ChIP-qPCR figure).
CUT & Tag Image 1: Bmi1 (D42B3) Rabbit mAb
Orders: 877-616-CELL (2355) • [email protected] • Support: 877-678-TECH (8324) • [email protected] • Web: cellsignal.com
For Research Use Only. Not for Use in Diagnostic Procedures.

Revision 6

#5856

Bmi1 (D42B3) Rabbit mAb

Cell Signaling Technology Logo
CUT&Tag was performed with NCCIT cells and Bmi1 (D42B3) Rabbit mAb, using CUT&Tag Assay Kit #77552. DNA library was prepared using CUT&Tag Dual Index Primers and PCR Master Mix for Illumina Systems #47415. The figures show binding across the HOXA gene cluster (upper), and the HOXD gene cluster (lower), which are known target genes of Bmi1 (see our ChIP-qPCR figure).
CUT & Tag Image 2: Bmi1 (D42B3) Rabbit mAb
Orders: 877-616-CELL (2355) • [email protected] • Support: 877-678-TECH (8324) • [email protected] • Web: cellsignal.com
For Research Use Only. Not for Use in Diagnostic Procedures.