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Antibodies

Histone H3K27me1 antibody (mAb)

Clone: MABI 0321
Catalog No: 61015 Format: 100 µg $420 Buy Now

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Antibody Type:Monoclonal
Isotype:IgG2a
Purification:Protein G Chromatography
Host:Mouse
Molecular Weight:17 kDa
Reactivity:Human, Other (Wide Range)

Applications

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

for Histone H3 monomethyl Lys27 antibody (mAb) (Clone MABI 0321)Application Notes


Validated Applications:
ChIP: 2 - 5 µg per ChIP
ChIP-Seq: 2 - 5 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
ICC/IF
WB
See references for more information. Individual optimization may be required.

for Histone H3 monomethyl Lys27 antibody (mAb) (Clone MABI 0321)Immunogen

This Histone H3 monomethyl Lys27 antibody was raised against a peptide containing monomethyl Lys27 of human Histone H3.

for Histone H3 monomethyl Lys27 antibody (mAb) (Clone MABI 0321)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 monomethyl Lys27 antibody (mAb) (Clone MABI 0321)References

 
Histone H3K27me1 antibody (mAb) tested by ChIP-Seq.

Histone H3K27me1 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 B cell lymphoma cell line and 2 µg of antibody. ChIP DNA was sequenced on the Illumina HiSeq and 15 million sequence tags were mapped to identify Histone H3K27me1 binding sites. The image shows binding across a region of chromosome 6. You can view the complete data set in the UCSC Genome Browser, starting at this specific location, here.

Histone H3K27me1 antibody (mAb) tested by ChIP.

Histone H3K27me1 antibody (mAb) tested by ChIP.
Chromatin IP performed using the ChIP-IT® Express Kit (Catalog No. 53008) and HeLa Chromatin (1.5 x 106 cell equivalents per ChIP) using 10 µg of Histone H3 monomethyl Lys27 pAb or the equivalent amount of rabbit IgG. RT-qPCR was performed on DNA purified from each of the ChIP DNA using a primer pair for the MyoD gene. Data are presented as Fold Enrichment of the antibody signal versus the negative control IgG using the ddCT method.

Histone H3K27me1 antibody (mAb) tested by Western blot.

Histone H3K27me1 antibody (mAb) tested by Western blot.
HeLa nuclear extract (20 µg per lane) probed with Histone H3 monomethyl Lys27 antibody (2 µg/ml dilution).

Histone H3K27me1 antibody (mAb) tested by dot blot analysis.

Histone H3K27me1 antibody (mAb) tested by dot blot analysis.
Dot blot analysis was used to confirm the specificity of Histone H3 monomethyl Lys27 antibody for monomethyl Lys27 of histone H3. Recombinant methylated histone proteins corresponding to the immunogen and related sequences were spotted onto PVDF and probed with Histone H3 monomethyl Lys27 at 2 µg/ml. The amount of protein (picomoles) spotted is indicated next to each row. Lane 1: unmodified H3 protein. Lane 2: monomethyl Lys4 protein. Lane 3: dimethyl Lys4 protein. Lane 4: trimethyl Lys4 protein. Lane 5: monomethyl Lys9 protein. Lane 6: dimethyl Lys9 protein. Lane 7: trimethyl Lys9 protein. Lane 8: monomethyl Lys27 protein. Lane 9: dimethyl Lys27 protein. Lane 10: trimethyl Lys27 protein.

for Histone H3 monomethyl Lys27 antibody (mAb) (Clone MABI 0321)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). Histone H1 is a linker histone, present at the interface between the nucleosome core and DNA entry/exit points. Histone H1 is responsible for establishing higher-order chromatin structure. 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. Lysine 27 of histone H3 can be mono-, di- or trimethylated (Histone H3 monomethyl Lys27, Histone H3 dimethyl Lys27 or Histone H3 trimethyl Lys27) by different histone methyltransferases such as EZH2 or NSD3. Methylation of this residue is mainly associated with transcriptional repression.

for Histone H3 monomethyl Lys27 antibody (mAb) (Clone MABI 0321)Positive Control

Active Motif's HeLa nuclear extract (Catalog No. 36010) can be used as a positive control for Histone H3 monomethyl Lys27 antibody.

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 H3K27me1 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 H3K27me1 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 H3K27me1 antibody (mAb).

for Histone H3 monomethyl Lys27 antibody (mAb) (Clone MABI 0321)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 monomethyl Lys27 antibody (mAb) (Clone MABI 0321)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 H3K27me1 antibody (mAb)
 

Data Thumbnails

Histone H3K27me1 antibody (mAb) tested by ChIP-Seq.

ChIP-Seq of Histone H3K27me1 mAb.
(Click image to enlarge and see details.)

Histone H3K27me1 antibody (mAb) tested by ChIP.

ChIP of Histone H3K27me1 mAb.
(Click image to enlarge and see details.)

Histone H3K27me1 antibody (mAb) tested by Western blot.

Western blot of Histone H3K27me1 mAb.
(Click image to enlarge and see details.)

Histone H3K27me1 antibody (mAb) tested by dot blot analysis.

Dot blot of Histone H3K27me1 mAb.
(Click image to enlarge and see details.)