Histone H3K14ac antibody (pAb)

RRID: AB_2793310
Catalog No: 39697 Format: 100 µg Discontinued
Catalog No: 39698 Format: 10 µg Discontinued

Antibody Type:
Polyclonal
Isotype:
IgG
Purification:
Protein A Chromatography
Host:
Rabbit
Molecular Weight:
17 kDa
Reactivity:
Human, Wide Range Predicted

Applications

ChIP Validated Western Blot Validated Dot Blot Validated

Application Notes

Applications Validated by Active Motif:
ChIP: 2 - 5 µg per ChIP
WB: 0.5 - 2 µg/ml dilution

By dot blot, slight reactivity towards histone H3 acetyl Lys23 might be observed under certain conditions. Individual optimization may be required.
This product has been discontinued:
Please refer to our alternative product Catalog No. 65701, Catalog No. 91397

Immunogen

This Histone H3 acetyl Lys14 antibody was raised against a peptide including acetyl-lysine 14 of histone H3.

Buffer

Purified IgG in PBS (pH 7.5) with 30% glycerol and 0.035% sodium azide. Sodium azide is highly toxic. For your convenience, an unpurified serum version (Catalog No. 39599) of this antibody is also available.

View our Guide to Histone Modifications and Biological Function.

 

Histone H3 acetyl Lys14 pAb tested by ChIP analysis.
Chromatin IP performed using the ChIP-IT® Express Kit (Catalog No. 53008) and HeLa Chromatin (1.5 x 106 cell equivalents per ChIP) using 5 µg of Histone H3 acetyl Lys14 pAb or the equivalent amount of rabbit IgG as a negative control. Real time, quantitative PCR (RT-qPCR) was performed on DNA purified from each of the ChIP reactions using a primer pair specific for the indicated gene. Data are presented as Fold Enrichment of the ChIP antibody signal versus the negative control IgG using the ddCT method.

Histone H3 acetyl Lys14 pAb tested by Western blot.
HeLa acid extract (10 µg per lane) was probed with Histone H3 acetyl Lys14 pAb (2 µg/ml dilution).
Lane 1: No treatment.
Lane 2: Cells treated with sodium butyrate.

Histone H3 acetyl Lys14 pAb tested by dot blot analysis.
Dot blot analysis was used to confirm the specificity of Histone H3 acetyl Lys14 pAb for acetyl Lys14 histone H3. Acetylated peptides corresponding to the immunogen and related peptides were spotted onto PVDF and probed with the antibody at a dilution of 1 µg/ml. The amount of peptide (picomoles) spotted is indicated next to each row.
Lane 1: acetyl-Lys9 peptide.     Lane 2: unmodified Lys9 peptide.
Lane 3: acetyl-Lys14 peptide.   Lane 4: unmodified Lys14 peptide.
Lane 5: acetyl-Lys18 peptide.   Lane 6: acetyl-Lys23 peptide.
Lane 7: acetyl-Lys27 peptide.
No detection of peptides acetylated at either lysine 4, 36, 37, 64 or 79 of histone H3 was observed (data not shown).

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. Acetylation of histones is linked to a number of specific processes including transcriptional regulation and genomic organization.

Positive Control

Active Motif's HeLa acid extract (Sodium butyrate treated) (Catalog No. 36202) can be used as a positive control for Histone H3 acetyl Lys14 pAb.

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 H3K14ac antibody (pAb).

Storage

Some products may be shipped at room temperature. This will not affect their stability or performance. Avoid repeated freeze/thaw cycles by aliquoting items into single-use fractions for storage at -20°C for up to 2 years. Keep all reagents on ice when not in storage.

Guarantee

This product is guaranteed for 12 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;
  • ChIP = ChIP Sequencing;
  • CUT&RUN = Cleavage Under Targets and Release Using Nuclease;
  • CUT&Tag = Cleavage Under Targets and Tagmentation;
  • 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;
  • NEU = Neutralizing;
  • WB = Western Blotting