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Antibodies

Histone H3K36me3 antibody (pAb)

Catalog No: 61101 Format: 100 µg $420 Buy Now
Catalog No: 61102 Format: 10 µl $95 Buy Now

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Antibody Type:Polyclonal
Isotype:IgG
Purification:Protein A Chromatography
Host:Rabbit
Molecular Weight:17 kDa
Reactivity:Human, Mouse, Other (Wide Range)

Applications

ChIP Validated ChIP-Seq Validated Western Blot Validated Immunofluorescence Validated Dot Blot Validated Immunocytochemistry Validated

for Histone H3 trimethyl Lys36 antibody (pAb)Application Notes


Validated Applications:
ChIP: 10 µg per ChIP
ChIP-Seq: 4 µg each
ICC/IF: 1 µg/ml dilution
WB: 0.1 - 1 µg/ml dilution

Published Applications:
ChIP-Seq
ChIP
WB

for Histone H3 trimethyl Lys36 antibody (pAb)Immunogen

This Histone H3 trimethyl Lys36 antibody was raised against a peptide containing trimethyl Lys36 of human histone H3.

for Histone H3 trimethyl Lys36 antibody (pAb)Buffer

Purified IgG in PBS with 30% glycerol and 0.035% sodium azide. Sodium azide is highly toxic.

for Histone H3 trimethyl Lys36 antibody (pAb)References

 
Histone H3K36me3 antibody (pAb) tested by ChIP-Seq.

Histone H3 trimethyl Lys36 antibody (pAb) tested by ChIP-Seq.
ChIP was performed using the ChIP-IT® High Sensitivity Kit (Cat. No. 53040) with 30 µg of chromatin from a human kidney. ChIP DNA was sequenced on the Illumina GA II and 30 million sequence tags were mapped to identify H3K36me3 binding across the genome. The image is zoomed in to show H3K36me3 binding across 5 different genes on chromosome 20.

Histone H3K36me3 antibody (pAb) tested by ChIP.

Chromatin IP:
ChIP performed using HeLa Chromatin (1.5 x 106 cell equivalents per ChIP) and 10 µg of Histone H3 trimethyl Lys36 antibody (pAb) or the equivalent amount of rabbit IgG as a negative control. Real time, quantitative PCR (RT-qPCR) was performed on DNA purified from each of the ChIP reactions using a primer pair specific for either the PABPC1 (+ 0.5) or the PABPC1 ( +18) gene. Data are presented as Fold Enrichment of the ChIP antibody signal versus the negative control IgG using the ddCT method.

Histone H3K36me3 antibody (pAb) tested by immunofluorescence.

Histone H3K36me3 antibody (pAb) tested by immunofluorescence.
HeLa cells stained with Histone H3K36me3 antibody (green) at a 1 ug/ml dilution. Tubulin staining was done with our alpha Tubulin antibody, catalog no.39527 (red) at a 1:1,000 dilution.

Histone H3K36me3 antibody (pAb) tested by Western blot.

Western Blot:
Nuclear extract of HeLa cells (20 µg) probed with the Histone H3 trimethyl Lys36 antibody at a dilution of 1 µg/ml.

Histone H3K36me3 antibody (pAb) tested by dot blot analysis.

Specificity Data:
Dot blot analysis was used to confirm the specificity of Histone H3 trimethyl Lys36 antibody for trimethyl-lysine 36 of histone H3. Peptides corresponding to regions around major sites of histone H3 methylation were spotted onto PVDF and probed with the antibody at a dilution of 1 µg/ml. The amount of peptide (in picomoles) spotted is indicated next to each row. Top panel - Lane 1: unmodified Lys4. Lane 2: monomethyl Lys4. Lane 3: dimethyl Lys4. Lane 4: trimethyl Lys4. Lane 5: unmodified Lys9. Lane 6: monomethyl Lys9. Lane 7: dimethyl Lys9. Lane 8: trimethyl Lys9. Lane 9: unmodified Lys79. Lane 10: monomethyl Lys79. Lane 11: dimethyl Lys79. Lane 12: trimethyl Lys79. Bottom panel - Lane 1: Unmodified Lys23. Lane 2: Monomethyl Lys23. Lane 3: Dimethyl Lys23. Lane 4: Trimethyl Lys23. Lane 5: unmodified Lys27. Lane 6: monomethyl Lys27. Lane 7: dimethyl Lys27. Lane 8: trimethyl Lys27. Lane 9: unmodified Lys36. Lane 10: monomethyl Lys36. Lane 11: dimethyl Lys36. Lane 12: trimethyl Lys36.

for Histone H3 trimethyl Lys36 antibody (pAb)Background

Histone H3 is one of the core components of the nucleosome. The nucleosome is the smallest subunit of chromatin and consists of 147 base pairs of DNA wrapped around an octamer of core histone proteins (two each of Histone H2A, Histone H2B, Histone H3 and Histone H4). Chromatin is subject to a variety of chemical modifications, including post-translational modifications of the histone proteins and the methylation of cytosine residues in the DNA. Reported histone modifications include acetylation, methylation, phosphorylation, ubiquitylation, glycosylation, ADP-ribosylation, carbonylation and SUMOylation; these modifications play a major role in regulating gene expression. The methylation of histones can occur on two different residues: arginine or lysine. Histone methylation can be associated with transcriptional activation or repression, depending on the methylated residue. Histone H3 is methylated at lysine 36 by the Set2 (yeast) and NSD1 (mammals) methyltransferases. Dimethylation of lysine 36 of histone H3 is involved with transcriptional elongation by RNA pol II holoenzyme and is a marker of transcribed genes.

for Histone H3 trimethyl Lys36 antibody (pAb)Positive Control

Active Motif's HeLa nuclear extract (Catalog No. 36010) can be used as a positive control for Histone H3 trimethyl Lys36 antibody.

for Histone H3 trimethyl Lys36 antibody (pAb)qPCR Primers

Active Motif's Human Positive Control Primer Set ACTB-1 (Cat. No. 71003) and Human Negative Control Primer Set 1 (Cat. No. 71001) have been validated for performing qPCR and endpoint PCR when this antibody has been used for ChIP on human samples. For ChIP experiments with this antibody on mouse samples, the Mouse Positive Control Primer Set Actb-2 (Cat. No. 71017) and Mouse Negative Control Primer Set 1 (Cat. No. 71011) have been validated for both qPCR and endpoint PCR.

Chromatin IP Assays

This antibody has been validated for use in ChIP and/or ChIP-Seq, and can be used with Active Motif's ChIP-IT® High Sensitivity Kit or our magnetic bead-based ChIP-IT® Express Kits. For an overview of all of our ChIP products, please visit our Chromatin IP Page to learn about ChIP products designed for use with Histone H3K36me3 antibody (pAb).

Compatible Secondary Antibodies

Active Motif has developed a variety of high-quality secondary antibodies for most applications, with anti-rabbit and anti-mouse conjugates to a broad line of high-quality fluorescent molecules, as well as horse radish peroxidase (HRP). Visit our Secondary Antibody Conjugates page to find the perfect secondaries for immunofluorescence experiments with Histone H3K36me3 antibody (pAb).

for Histone H3 trimethyl Lys36 antibody (pAb)Storage

Some products may be shipped at room temperature. This will not affect their stability or performance. Upon receipt, unconjugated antibodies may be stored at -20°C for up to 2 years. Fluorophore- & enzyme-conjugated antibodies should be stored at 4°C. Fluorophore-conjugated antibodies should be protected from light. Keep reagents on ice when not in storage; to avoid repeated freeze/thaw cycles, we recommend aliquoting items that will be stored frozen into single-use fractions prior to freezing.

for Histone H3 trimethyl Lys36 antibody (pAb)Guarantee

This product is guaranteed for 6 months from date of receipt.

This product is for research use only and is not for use in diagnostic procedures.

Application Key

  • ChIP = Chromatin Immunoprecipitation;
  • DB = Dot Blot;
  • ELISA = Enzyme-linked Immunosorbent Assay;
  • EMSA = Electrophoretic Mobility Shift Assay
  • FC = Flow Cytometry;
  • ICC = Immunocytochemistry;
  • IF = Immunofluorescence;
  • IHC = Immunohistochemistry;
  • IP = Immunoprecipitation;
  • MeDIP = Methyl-DNA Immunoprecipitation;
  • TR-FRET = Time-Resolved Fluorescence Resonance Energy Transfer;
  • WB = Western Blotting
 

Technical Data Sheet

Histone H3K36me3 antibody (pAb)
 

Data Thumbnails

Histone H3K36me3 antibody (pAb) tested by ChIP-Seq.

ChIP-Seq of Histone H3K36me3 pAb.
(Click image to enlarge and see details.)

Histone H3K36me3 antibody (pAb) tested by ChIP.

ChIP of Histone H3K36me3 pAb.
(Click image to enlarge and see details.)

Histone H3K36me3 antibody (pAb) tested by immunofluorescence.

Immunofluorescence stain of Histone H3K36me3 pAb.
(Click image to enlarge and see details.)

Histone H3K36me3 antibody (pAb) tested by Western blot.

Western blot of Histone H3K36me3 pAb.
(Click image to enlarge and see details.)

Histone H3K36me3 antibody (pAb) tested by dot blot analysis.

Dot blot of Histone H3K36me3 pAb.
(Click image to enlarge and see details.)