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Antibodies

Histone H3K36me3 antibody (mAb)

Clone: MABI 0333
Catalog No: 61021 Format: 100 µg $420 Buy Now

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Antibody Type:Monoclonal
Isotype:IgG1
Purification:Protein G Chromatography
Host:Mouse
Molecular Weight:17 kDa
Reactivity:Human, Mouse, Other (Wide Range)

Applications

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

for Histone H3 trimethyl Lys36 antibody (mAb) (Clone MABI 0333)Application Notes


Validated Applications:
ChIP: 5 - 10 µg per ChIP
ChIP-Seq: 5 - 10 µg each
WB: 0.5 - 2 µg/ml dilution

ChIP-Seq validation was performed by Active Motif's Epigenetics Services and the complete data set is available in the UCSC Genome Browser by clicking here.

Published Applications:
ChIP
WB
IF
See references for more information. Individual optimization may be required.

for Histone H3 trimethyl Lys36 antibody (mAb) (Clone MABI 0333)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 (mAb) (Clone MABI 0333)Buffer

PBS pH 7.5 containing 30% glycerol and 0.035% sodium azide. Sodium azide is highly toxic. For your convenience, a PBS only version of this antibody is also available.

for Histone H3 trimethyl Lys36 antibody (mAb) (Clone MABI 0333)References

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

Histone H3K36me3 antibody (mAb) tested by ChIP-Seq.
ChIP was performed using the ChIP-IT® High Sensitivity Kit (Cat. No. 53040) with 15 ug of chromatin from a human medulloblastoma cell line and 4 µg of antibody. ChIP DNA was sequenced on the Illumina HiSeq and 15 million sequence tags were mapped to identify Histone H3K36me3 binding sites. The image shows binding across a region of chromosome 7. You can view the complete data set in the UCSC Genome Browser, starting at this specific location, here.

Histone H3K36me3 antibody (mAb) tested by Western blot.

Histone H3K36me3 antibody (mAb) tested by Western blot
HeLa nuclear extract (20 µg per lane) probed with Histone H3 trimethyl Lys36 antibody (2 µg/ml dilution).

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

Histone H3K36me3 antibody (mAb) tested by dot blot.
Dot blot analysis was used to confirm the specificity of Histone H3K36me3 antibody for trimethyl Lys36 of histone H3. Recombinant methylated proteins corresponding to the immunogen and related sequences were spotted onto PVDF and probed with Histone H3K36me3 at 2 µg/ml. The amount of protein (picomoles) spotted is indicated next to each row.
Top panel - Lane 1: unmodified H3 Lys4. Lane 2: H3K4me1. Lane 3: H3K4me2. Lane 4: H3K4me3. Lane 5: unmodified H3 Lys9. Lane 6: H3K9me1. Lane 7: H3K9me2. Lane 8: H3K9me3. Lane 9: unmodified H3 Lys79. Lane 10: H3K79me1. Lane 11: H3K79me2. Lane 12: H3K79me3. Bottom panel - Lane 1: unmodified H3 Lys23. Lane 2: H3K23me1. Lane 3: H3K23me2. Lane 4: H3K23me3. Lane 5: unmodified H3 Lys27. Lane 6: H3K27me1. Lane 7: H3K27me2. Lane 8: H3K27me3. Lane 9: unmodified H3 Lys36. Lane 10: H3K36me1. Lane 11: H3K36me2. Lane 12: H3K36me3.

for Histone H3 trimethyl Lys36 antibody (mAb) (Clone MABI 0333)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 (mAb) (Clone MABI 0333)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 (mAb) (Clone MABI 0333)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-1 (Cat. No. 71015) 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 (mAb).

Bridging Antibody to Improve ChIP, MeDIP and IP

Some isotypes of mouse IgG antibodies do not bind well to protein G-conjugated agarose/magnetic beads, which are commonly used in ChIP, MeDIP and IP experiments. Capture efficiency can be improved by including an anti-mouse IgG bridging antibody because it binds with a strong affinity to both the protein G beads and the mouse IgG antibody. By increasing the affinity of the mouse primary antibody for the protein G bead, the Bridging Antibody for Mouse IgG can help improve the results of all ChIP, MeDIP and IP experiments that use Histone H3K36me3 antibody (mAb).

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 (mAb).

for Histone H3 trimethyl Lys36 antibody (mAb) (Clone MABI 0333)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 (mAb) (Clone MABI 0333)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.

This antibody is manufactured by MAB Institute, Inc.

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 (mAb)
 

Data Thumbnails

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

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

Histone H3K36me3 antibody (mAb) tested by Western blot.

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

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

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