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Antibodies

AbFlex® Histone H3K36me3 antibody (rAb)

Catalog No: 91265 Format: 100 µg $375 Buy Now
Catalog No: 91266 Format: 10 µg $75 Buy Now

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Antibody Type:Recombinant
Isotype:IgG2a
Purification:Protein A Chromatography
Host:Mouse
Molecular Weight:17 kDa
Reactivity:Other (Wide Range)

Applications

ChIP Validated ChIP-Seq Validated Western Blot Validated

for AbFlex Histone H3K36me3 antibody (rAb)Application Notes


Validated Applications:
ChIP-Seq: 4 µg per ChIP
WB: 0.5 - 2 µg / mL

for AbFlex Histone H3K36me3 antibody (rAb)Immunogen

This antibody was raised against a peptide containing trimethly-lysine 36 of histone H3.

for AbFlex Histone H3K36me3 antibody (rAb)Buffer

Purified IgG in 140 mM Hepes, pH 7.5, 70 mM NaCl, 32 mM NaOAc, 0.035% sodium azide, 30% glycerol.

View our Guide to Histone Modifications and Biological Function.

 
AbFlex<sup>®</sup> Histone H3K36me3 antibody (rAb) tested by ChIP-Seq.

AbFlex® Histone H3K36me3 recombinant antibody (rAb) tested by ChIP-Seq
Chromatin immunoprecipitation (ChIP) was performed using the ChIP-IT® High Sensitivity Kit (Cat. No. 53040) with 30 µg of HAP1 cell line- myeloid leukemia chromatin and 4 µg of Histone H3K36me3 antibody. ChIP DNA was sequenced on the Illumina NextSeq and 15.4 million sequence tags were mapped to identify Histone H3K36me3 binding sites on chromosome 16. You can view the complete data set in the UCSC Genome Browser, starting at this specific location, here.

AbFlex<sup>®</sup> Histone H3K36me3 antibody (rAb) tested by Western blot.

AbFlex® Histone H3K36me3 recombinant antibody (rAb) tested by Western blot
20 µg of HeLa cell nuclear extract (Lane 1) and 15 µg HeLa whole cell acid extract(Lane 2) were run on SDS-PAGE and probed with AbFlex H3K36me3 antibody at 0.5 µg/ml.

Histone H3K36me3 antibody (rAb) specificity tested by peptide array analysis.

Histone H3K36me3 antibody (rAb) specificity tested by peptide array analysis.
Peptide array analysis was used to confirm the specificity of this antibody for its intended modification. Histone H3K36me3 antibody was applied at a dilution of 0.2 µg/ml to Active Motif's MODified™ Histone Peptide Array (Catalog No. 13001). The arrays were scanned with ArrayAnalysis Software 16 and the results plotted. Specificity data is shown for the most reactive peptides and those related to the immunogen
Array Data File

for AbFlex Histone H3K36me3 antibody (rAb)Background

AbFlex® antibodies are recombinant antibodies (rAbs) that have been generated using defined DNA sequences to produce highly specific, reproducible antibodies. Each AbFlex antibody contains a 6XHis tag, an avidin tag sequence for enzymatic biotin conjugation using the biotin ligase, BirA, and a sortase recognition motif (LPXTG) to attach a variety of labels directly to the antibody including fluorophores, enzymatic substrates (HRP, AP), peptides, drugs as well as solid supports. AbFlex H3K36me3 antibody was expressed as full-length IgG with mouse immunoglobulin heavy and light chains (IgG2a isotype) in mammalian 293 cells.

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.

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 AbFlex® Histone H3K36me3 antibody (rAb).

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 AbFlex® Histone H3K36me3 antibody (rAb).

for AbFlex Histone H3K36me3 antibody (rAb)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 AbFlex Histone H3K36me3 antibody (rAb)Guarantee

This product is for research use only and is not for use in diagnostic procedures. This product is guaranteed for 6 months from date of arrival.

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

AbFlex® Histone H3K36me3 antibody (rAb)
 

Data Thumbnails

AbFlex<sup>®</sup> Histone H3K36me3 antibody (rAb) tested by ChIP-Seq.

ChIP-Seq of AbFlex® Histone H3K36me3 antibody (rAb).
(Click image to enlarge and see details.)

AbFlex<sup>®</sup> Histone H3K36me3 antibody (rAb) tested by Western blot.

Western blot of AbFlex® Histone H3K36me3 antibody (rAb).
(Click image to enlarge and see details.)

Histone H3K36me3 antibody (rAb) specificity tested by peptide array analysis.

Histone H3K36me3 antibody (rAb) specificity tested by peptide array analysis.
(Click to enlarge and view details)