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Mono-Methyl-Histone H3 (Lys4) (D1A9) XP ® Rabbit mAb
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Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to the amino terminus of histone H3 in which lysine 4 is mono-methylated.
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 |
| Immunofluorescence (Immunocytochemistry) | 1:400 - 1:1600 |
| Flow Cytometry (Fixed/Permeabilized) | 1:200 - 1:800 |
| Chromatin IP | 1:50 |
| Chromatin IP-seq | 1:50 |
| 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 #53138.
参考图片
CUT&Tag was performed with HeLa cells and Mono-Methyl-Histone H3 (Lys4) (D1A9) 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 chromosome 8 (upper), including ADAM9 (lower), a known target gene of H3K4me1 (see our ChIP-qPCR figure).
Western blot analysis of extracts from various cell lines using Mono-Methyl-Histone H3 (Lys4) (D1A9) XP® Rabbit mAb.
Confocal immunofluorescent analysis of HeLa cells using Mono-Methyl-Histone H3 (Lys4) (D1A9) XP® Rabbit mAb (green) and MEK1/2 (L38C12) Mouse mAb #4694 (blue). Actin filaments were labeled with Dy-554 phalloidin (red).
Flow cytometric analysis of HeLa cells using Mono-Methyl-Histone H3 (Lys4) (D1A9) 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® 647 Conjugate) #4414 was used as a secondary antibody.
Chromatin immunoprecipitations were performed with cross-linked chromatin from Hela cells and Mono-Methyl-Histone H3 (Lys4) (D1A9) XP® Rabbit mAb, using SimpleChIP® Enzymatic Chromatin IP Kit (Magnetic Beads) #9003. DNA Libraries were prepared using DNA Library Prep Kit for Illumina® (ChIP-seq, CUT&RUN) #56795. The figure shows binding across ADAM9, a known target gene of H3K4me1 (see additional figure containing ChIP-qPCR data).
Chromatin immunoprecipitations were performed with cross-linked chromatin from Hela cells and Mono-Methyl-Histone H3 (Lys4) (D1A9) XP® Rabbit mAb, using SimpleChIP® Enzymatic Chromatin IP Kit (Magnetic Beads) #9003. DNA Libraries were prepared using DNA Library Prep Kit for Illumina® (ChIP-seq, CUT&RUN) #56795. The figure shows binding across chromosome 8 (upper), including ADAM9 (lower), a known target gene of H3K4me1 (see additional figure containing ChIP-qPCR data).
Chromatin immunoprecipitations were performed with cross-linked chromatin from K562 cells and either Mono-Methyl-Histone H3 (Lys4) (D1A9) 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 ADAM9 Intron 11 Primers #73401, human ADAM18 intron 14 primers, human Trio inton 1 primers, and SimpleChIP® Human Trio Exon 57 Primers #90568. 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 was performed with HeLa cells and Mono-Methyl-Histone H3 (Lys4) (D1A9) XP® Rabbit mAb using CUT&RUN Assay Kit #86652. DNA library was prepared using DNA Library Prep Kit for Illumina (ChIP-seq, CUT&RUN) #56795. The figures show binding across Adam9, a known target gene of Mono-Methyl-Histone H3 (Lys4) (see additional figure containing qPCR data).
CUT&RUN was performed with HeLa cells and Mono-Methyl-Histone H3 (Lys4) (D1A9) XP® Rabbit mAb using CUT&RUN Assay Kit #86652. DNA library was prepared using DNA Library Prep Kit for Illumina (ChIP-seq, CUT&RUN) #56795. The figures show binding across chromosome 16 (upper), including Adam9 (lower), a known target gene of Mono-Methyl-Histone H3 (Lys4) (see additional figure containing qPCR data).
CUT&RUN was performed with HeLa cells and either Mono-Methyl-Histone H3 (Lys4) (D1A9) 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 ADAM9 Intron 11 Primers #73401, human Trio intron 1 primers, and human ADAM18 intron 14 primers. 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 HeLa cells and Mono-Methyl-Histone H3 (Lys4) (D1A9) 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 ADAM9, a known target gene of H3K4me1 (see our ChIP-qPCR figure).