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Antibodies

Histone H3K4me2 antibody (mAb)

Clone: MABI 0303
Catalog No: 39679 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 Immunocytochemistry Validated

for Histone H3 dimethyl Lys4 antibody (mAb) (Clone MABI 0303)Application Notes


Validated Applications:
ChIP: 5 - 10 µg per ChIP
ChIP-Seq: 5 - 10 µg each
ICC/IF: 1 - 2 µl/ml dilution
WB: 2 - 5 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
See references for more information. Individual optimization may be required.

for Histone H3 dimethyl Lys4 antibody (mAb) (Clone MABI 0303)Immunogen

This Histone H3 dimethyl Lys4 antibody was raised against a synthetic peptide containing dimethyl Lys4 of human histone H3.

for Histone H3 dimethyl Lys4 antibody (mAb) (Clone MABI 0303)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 dimethyl Lys4 antibody (mAb) (Clone MABI 0303)References

 
Histone H3K4me2 antibody (mAb) tested by ChIP-Seq.

Histone H3K4me2 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 12 million sequence tags were mapped to identify Histone H3K4me2 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 H3K4me2 antibody (mAb) tested by immunofluorescence.

Histone H3K4me2 antibody (mAb) tested by immunofluorescence.
Staining of HeLa cells with Histone H3 dimethyl Lys4 antibody (mAb) (1 µg/ml, top panel) and DAPI (middle panel), and a merge of both images (bottom panel).

Histone H3K4me2 antibody (mAb) tested by Histone Peptide Array

Histone H3K4me2 antibody (mAb) specificity tested by peptide array analysis.
Peptide array analysis was used to confirm the specificity of this antibody for its intended modification. Histone H3 dimethyl Lys4 antibody was applied at a dilution of 1:2,000 to Active Motif's MODified™ Histone Peptide Array (Catalog No. 13001). The arrays were scanned with ArrayAnalysis Software 7 and the results plotted. Specificity data is shown for the most reactive peptides and those related to the immunogen. Recognition of the H3 dimethyl Lys4 peptide is inhibited by Thr3 phosphorylation and blocked by a citrulline at position 2.
Array Data File

Histone H3K4me2 antibody (mAb) tested by Western blot.

Histone H3K4me2 antibody (mAb) tested by Western blot.
HeLa acid extract (20 µg) probed with Histone H3 dimethyl Lys4 antibody (mAb) (2 µg/ml dilution).

Histone H3K4me2 antibody (mAb) tested by dot blot analysis.

Histone H3K4me2 antibody (mAb) tested by dot blot analysis.
Dot blot analysis was used to confirm the specificity of Histone H3 dimethyl Lys4 antibody (mAb) for dimethyl-Lys4 histone H3. Methylated peptides corresponding to the immunogen and related peptides were spotted onto PVDF and probed with the antibody at 1 µg/ml. The amount of peptide (picomoles) spotted is indicated next to each row.
Top panel: Lane 1: Unmod H3 aa 2-9 peptide lysine 4. Lane 2: Monomethyl lysine 4. Lane 3: Dimethyl lysine 4. Lane 4: Trimethyl lysine 4. Lane 5: Unmod H3 5-22 peptide. Lane 6: Monomethyl lysine 9. Lane 7: Dimethyl lysine 9. Lane 8: Trimethyl lysine 9. Bottom panel: Lane 1: dimethyl lysine 14. Lane 2: monomethyl lysine 18. Lane 3: dimethyl lysine 18. Lane 4: trimethyl lysine 18. Lane 5: unmod H3 aa 18-27 peptide. Lane 6: monomethyl lysine 23. Lane 7: dimethyl lysine 23. Lane 8: trimethyl lysine 23. Lane 9: unmod H3 aa 22-32 peptide. Lane 10: monomethyl lysine 27. Lane 11: dimethyl lysine 27. Lane 12: trimethyl lysine 27.

for Histone H3 dimethyl Lys4 antibody (mAb) (Clone MABI 0303)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 4 of histone H3 can be mono-, di- or trimethylated by different histone methyltransferases (HMTs) such as SET1 or ASH1. Methylation of Lys4 is often associated with transcriptional activation. The demethylase LSD1 is able to demethylate histone H3 Lys4.

for Histone H3 dimethyl Lys4 antibody (mAb) (Clone MABI 0303)qPCR Primers

Active Motif's Human Positive Control Primer Set ACTB-2 (Cat. No. 71005) 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 H3K4me2 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 H3K4me2 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 H3K4me2 antibody (mAb).

for Histone H3 dimethyl Lys4 antibody (mAb) (Clone MABI 0303)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 dimethyl Lys4 antibody (mAb) (Clone MABI 0303)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 H3K4me2 antibody (mAb)
 

Data Thumbnails

Histone H3K4me2 antibody (mAb) tested by ChIP-Seq.

ChIP-Seq of Histone H3K4me2 mAb.
(Click image to enlarge and see details.)

Histone H3K4me2 antibody (mAb) tested by immunofluorescence.

Immunofluorescence stain of Histone H3K4me2 mAb.
(Click image to enlarge and see details.)

Histone H3K4me2 antibody (mAb) tested by Histone Peptide Array

Histone H3K4me2 antibody (mAb) tested by Histone Peptide Array
(Click image to enlarge and see details.)

Histone H3K4me2 antibody (mAb) tested by Western blot.

Western blot of Histone H3K4me2 mAb.
(Click image to enlarge and see details.)

Histone H3K4me2 antibody (mAb) tested by dot blot analysis.

Dot blot of Histone H3K4me2 mAb.
(Click image to enlarge and see details.)