Histone H3K14me3 antibody (pAb)

Catalog No: 61549 Format: 100 µl $435 Buy
Catalog No: 61550 Format: 10 µl $105 Buy

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Antibody Type:
Polyclonal
Isotype:
IgG
Purification:
Affinity Purified
Host:
Rabbit
Molecular Weight:
17 kDa
Reactivity:
Human, Other (Wide Range)

Applications

Western Blot Validated Dot Blot Validated

Application Notes


Validated Applications:
WB: 1:500 - 1:1,000 dilution

Note: many chromatin-bound proteins are not soluble in a low salt nuclear extract and fractionate to the pellet. Therefore, we recommend a High Salt / Sonication Protocol when preparing nuclear extracts for Western Blot.

Immunogen

This antibody was raised against a peptide including trimethyl lysine 14 of human Histone H3.

Buffer

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

View our Guide to Histone Modifications and Biological Function.

References

 
Histone H3K14me3 antibody (pAb) tested by Western blot.

Histone H3K14me3 antibody (pAb) tested by Western blot.
H3K14me3 detection by Western blot. The analysis was performed using 25 µg Jurkat nuclear cell extract and H3K14me3 antibody at a 1:500 dilution.

Histone H3K14me3 antibody (pAb) tested by dot blot analysis.

Histone H3K14me3 (pAb) tested by dot blot analysis.
Dot blot analysis was used to confirm the specificity of Histone H3K14me3 antibody (pAb) for trimethyl-lysine 14 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 1:1,000. The amount of peptide (in picomoles) spotted is indicated next to each row.Top panel - Lane 1: unmodified Lys14. Lane 2: monomethyl Lys14. Lane 3: dimethyl Lys14. Lane 4: trimethyl Lys14. Middle panel - Lane 1: unmodified Lys4. Lane 2: monomethyl Lys4. Lane 3: dimethyl Lys4. Lane 4: trimethyl Lys4. Lane 5: unmodified Lys9. Lane 6: monomethyl Lys9. Lane 7: dimethyl Lys9. Lane 8: trimethyl Lys9. Lane 9: unmodified Lys79. Lane 10: monomethyl Lys79. Lane 11: dimethyl Lys79. Lane 12: trimethyl Lys79. Bottom panel - Lane 1: Unmodified Lys23. Lane 2: Monomethyl Lys23. Lane 3: Dimethyl Lys23. Lane 4: Trimethyl Lys23. Lane 5: unmodified Lys27. Lane 6: monomethyl Lys27. Lane 7: dimethyl Lys27. Lane 8: trimethyl Lys27. Lane 9: unmodified Lys36. Lane 10: monomethyl Lys36. Lane 11: dimethyl Lys36. Lane 12: trimethyl Lys36.

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.

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.

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