Histone H3K36me3 antibody (pAb)

RRID: AB_2615073
Catalog No: 61101 Format: 100 µg $470 Buy
Catalog No: 61102 Format: 10 µg $115 Buy
Catalog No: 61902 Format: 50 µg $275 Buy

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

Applications

CUT&RUN Validated CUT&Tag Validated ChIP Validated ChIP-Seq Validated Western Blot Validated

Application Notes

Applications Validated by Active Motif:
ChIP: 10 µg per ChIP
ChIP-Seq: 4 µg each
WB: 0.5 - 2 µg/ml dilution
CUT&Tag: 1 µg per 50 µl reaction

For Histone H3K36me3, we also offer AbFlex® Histone H3K36me3 Recombinant Antibody (rAb). For details, see Catalog No. 91265.

Published Applications

The following applications have been published using this antibody. Unless noted above, Active Motif may not have validated the antibody for use in these applications:

ChIP-qPCR, ChIP-Seq
CUT&RUN
CUT&Tag
ICC/IF
WB

View publications that use Active Motif products & services here. Enter the product number(s) to see publications & applications that use this antibody.

Immunogen

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

Buffer

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

 

Histone H3K36me3 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 glioma cell line. ChIP DNA was sequenced on the Illumina NextSeq and 16.8 million sequence tags were mapped to identify H3K36me3 binding on chromosome 7. The antibody was used at 4 µg per 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.

Western blot of Histone H3K36me3 pAb.
Nuclear extract of HeLa cells (20 µg) probed with Histone H3K36me3 antibody at a dilution of 0.5 µg/ml.

Dot blot specificity analysis of Histone H3K26me3 pAb:
Dot blot analysis was used to confirm the specificity of Histone H3K36me3 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: K4me1. Lane 3: K4me1. Lane 4: K4me2. Lane 5: K4me3. Lane 6: unmodified Lys9. Lane 7: K9me1. Lane 8: K9me2. Lane 9: K9me3. Lower panel - Lane 1: unmodified Lys27. Lane 2: K27me1. Lane 3: K27me2. Lane 4: K27me3. Lane 5: unmodified Lys36. Lane 6: K36me1. Lane 7: K36me2. Lane 8: K36me3.

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.

Positive Control

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

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).

Storage

Some products may be shipped at room temperature. This will not affect their stability or performance. Avoid repeated freeze/thaw cycles by aliquoting items into single-use fractions for storage at -20°C for up to 2 years. Keep all reagents on ice when not in storage.

Guarantee

This product is guaranteed for 12 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;
  • ChIP = ChIP Sequencing;
  • CUT&RUN = Cleavage Under Targets and Release Using Nuclease;
  • CUT&Tag = Cleavage Under Targets and Tagmentation;
  • 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;
  • NEU = Neutralizing;
  • WB = Western Blotting