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Monoclonal antibody is produced by immunizing animals with recombinant protein specific to the carboxy terminus of human Bmi1 protein.
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 | 1:200 |
| 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 |
Specificity/Sensitivity
Species Reactivity:
Human, Monkey
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.
参考图片
CUT&Tag was performed with NCCIT cells and Bmi1 (D20B7) XP® 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 cluser (upper), and the HOXD gene cluster (lower), which are known target genes of Bmi1 (see our ChIP-qPCR figure).
Western blot analysis of extracts from various cell lines using Bmi1 (D20B7) XP® Rabbit mAb.
Immunohistochemical analysis of paraffin-embedded human lung carcinoma using Bmi1 (D20B7) XP® Rabbit mAb.
Immunohistochemical analysis of paraffin-embedded human lymphoma using Bmi1 (D20B7) XP® Rabbit mAb.
Confocal immunofluorescent analysis of COS-7 cells using Bmi1 (D20B7) XP® Rabbit mAb (green). Actin filaments were labeled with DY-554 phalloidin (red).
Flow cytometric analysis of K562 cells using Bmi1 (D20B7) XP® Rabbit mAb (solid line) compared to concentration-matched Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (dashed line). Anti-rabbit IgG (H+L), F(ab')2 Fragment (Alexa Fluor® 488 Conjugate) #4412 was used as a secondary antibody.
Chromatin immunoprecipitations were performed with cross-linked chromatin from NCCIT cells and Bmi1 (D20B7) XP® Rabbit mAb using SimpleChIP® Plus Enzymatic Chromatin IP Kit (Magnetic Beads) #9005. DNA Libraries were prepared using DNA Library Prep Kit for Illumina Systems (ChIP-seq, CUT&RUN) #56795. The figure shows binding across HoxA, a known target gene of Bmi1 (see additional figure containing ChIP-qPCR data).
Chromatin immunoprecipitations were performed with cross-linked chromatin from NCCIT cells and Bmi1 (D20B7) XP® Rabbit mAb, using SimpleChIP® Plus Enzymatic Chromatin IP Kit (Magnetic Beads) #9005. DNA Libraries were prepared using DNA Library Prep Kit for Illumina Systems (ChIP-seq, CUT&RUN) #56795. The figures show binding across HoxA (upper) and HoxD (lower), known target genes of Bmi1 (see additional figure containing ChIP-qPCR data).
Chromatin immunoprecipitations were performed with cross-linked chromatin from NCCIT cells and either Bmi1 (D20B7) XP® 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).
CUT&RUN was performed with NCCIT cells and either Bmi1 (D20B7) XP® Rabbit mAb or CBX4 (E6L7X) Rabbit mAb #30559, using CUT&RUN Assay Kit #86652. DNA libraries were prepared using DNA Library Prep Kit for Illumina Systems (ChIP-seq, CUT&RUN) #56795. The figure shows binding across HoxA1 gene, a known target gene of Bmi1 (see additional figure containing CUT&RUN-qPCR data).
CUT&RUN was performed with NCCIT cells and either Bmi1 (D20B7) XP® Rabbit mAb or CBX4 (E6L7X) Rabbit mAb #30559, using CUT&RUN Assay Kit #86652. DNA libraries were prepared using DNA Library Prep Kit for Illumina Systems (ChIP-seq, CUT&RUN) #56795. The figure shows binding across HoxA gene (upper) and HoxD gene (lower), known target genes of CBX4 and Bmi1 (see additional figure containing CUT&RUN-qPCR data).
CUT&RUN was performed with NCCIT cells and treatment and either Bmi1 (D20B7) XP® 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&Tag was performed with NCCIT cells and Bmi1 (D20B7) XP® 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).