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Ubiquityl-Histone H2A (Lys119) (D27C4) XP ® Rabbit mAb
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Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues near the carboxy terminus of human histone H2A protein in which Lys119 is mono-ubiquitinated.
Product Usage Information
For optimal ChIP and ChIP-seq results, use 5 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 | 1:2000 |
| Immunoprecipitation | 1:100 |
| Immunofluorescence (Immunocytochemistry) | 1:800 - 1:1600 |
| Flow Cytometry (Fixed/Permeabilized) | 1:800 - 1:3200 |
| Chromatin IP | 1:100 |
| Chromatin IP-seq | 1:100 |
| CUT&RUN | 1:50 |
| CUT&Tag | 1:50 |
Specificity/Sensitivity
Species Reactivity:
Human, Mouse, Rat, 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 #45772.
参考图片
CUT&Tag was performed with NCCIT cells and Ubiquityl-Histone H2A (Lys119) (D27C4) 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 HOXA genes (upper) and HOXD genes (lower), which are known target genes of H2AK119Ub (see our ChIP-qPCR figure).
Western blot analysis of extracts from various cell lines using Ubiquityl-Histone H2A (Lys119) (D27C4) XP® Rabbit mAb.
Confocal immunofluorescent analysis of HeLa cells using Ubiquityl-Histone H2A (Lys119) (D27C4) XP® Rabbit mAb (green) and COX IV (4D11-B3-E8) Mouse mAb #11967 (red).
Flow cytometric analysis of HeLa cells using Ubiquityl-Histone H2A (Lys119) (D27C4) 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 either Ubiquityl-Histone H2A (Lys119) (D27C4) XP® Rabbit mAb or RING1B (D22F2) XP® Rabbit mAb #5694, using SimpleChIP® Enzymatic Chromatin IP Kit (Magnetic Beads) #9005. DNA Libraries were prepared using SimpleChIP® ChIP-seq DNA Library Prep Kit for Illumina® #56795. H2AK119Ub and RING1B are known to associate with each other on chromatin. The figure shows binding of both H2AK119Ub and RING1B across HOXD genes.
Chromatin immunoprecipitations were performed with cross-linked chromatin from NCCIT cells and either Ubiquityl-Histone H2A (Lys119) (D27C4) XP® Rabbit mAb or RING1B (D22F2) XP® Rabbit mAb #5694, using SimpleChIP® Enzymatic Chromatin IP Kit (Magnetic Beads) #9005. DNA Libraries were prepared using SimpleChIP® ChIP-seq DNA Library Prep Kit for Illumina® #56795. H2AK119Ub and RING1B are known to associate with each other on chromatin. The figure shows binding of both H2AK119Ub and RING1B across HOXA genes (upper) and HOXD genes (lower), which are known target genes of H2AK119Ub and RING1B (see additional figure containing ChIP-qPCR data).
Chromatin immunoprecipitations were performed with cross-linked chromatin from NCCIT cells and either Ubiquityl-Histone H2A (Lys119) (D27C4) 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, SimpleChIP® Human GAPDH Exon 1 Primers #5516, 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 Ubiquityl-Histone H2A (Lys119) (D27C4) XP® Rabbit mAb, using CUT&RUN Assay Kit #86652. The DNA library was prepared using DNA Library Prep Kit for Illumina Systems (ChIP-seq, CUT&RUN) #56795. The figure shows binding across TMEM54 gene.
CUT&RUN was performed with NCCIT cells and Ubiquityl-Histone H2A (Lys119) (D27C4) XP® Rabbit mAb, using CUT&RUN Assay Kit #86652. The DNA library was prepared using DNA Library Prep Kit for Illumina Systems (ChIP-seq, CUT&RUN) #56795. The figures show binding across TMEM54 gene (upper) and SEMA6C gene (lower).
CUT&RUN was performed with NCCIT cells and either Ubiquityl-Histone H2A (Lys119) (D27C4) 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, SimpleChIP® Human HoxA2 Promoter Primers, 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 Ubiquityl-Histone H2A (Lys119) (D27C4) 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 HOXD genes, which are known target genes of H2AK119Ub (see our ChIP-qPCR figure).