Histone H4K12ac antibody (pAb)

Catalog No: 39165 Format: 100 µl $445 Buy
Catalog No: 39066 Format: 50 µl $260 Buy
Catalog No: 39166 Format: 10 µl $105 Buy

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Antibody Type:
Polyclonal
Isotype:
Serum
Purification:
None
Host:
Rabbit
Molecular Weight:
8 kDa
Reactivity:
Budding Yeast, Human, Mouse, Other (Wide Range)

Applications

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

Application Notes


Validated Applications:
ChIP: 2 - 8 µl per ChIP
ChIP-Seq: 5 µl each
ICC/IF: 1:500 - 1:2,000 dilution
WB: 1:5,000 - 1:20,000 dilution

Published Applications:
ChIP / ChIP-Seq
WB
IF

The modENCODE and NIH Roadmap Epigenomics Mapping Consortiums have implemented rigorous standardization criteria for all assays and reagents to be used. As part of this initiative, antibody specificity testing and the ability of the antibodies to work in ChIP-Seq were assessed in a large-scale study. This Histone H4 acetyl Lys12 antibody was validated for ChIP-Seq in the study (see reference).

Immunogen

This Histone H4 acetyl Lys12 antibody was raised against a peptide including acetyl-lysine 12 of histone H4.

Buffer

Rabbit serum containing 30% glycerol and 0.035% sodium azide. Sodium azide is highly toxic. For your convenience, an IgG version (Catalog No. 39927) of this antibody that was purified by Protein A Chromatography is also available.

References

 
Histone H4K12ac antibody (pAb) tested by ChIP.

Histone H4 acetyl Lys12 antibody 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 10 µl of Histone H4 acetyl Lys12 antibody 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 H4K12ac antibody (pAb) tested by immunofluorescence.

Histone H4 acetyl Lys12 antibody tested by immunofluorescence.
Staining of HeLa cells with Histone H4 acetyl Lys12 antibody (1:500 dilution, top panel) and DAPI (bottom panel).

Histone H4K12ac antibody (pAb) tested by Western blot.

Histone H4 acetyl Lys12 antibody tested by Western blot.
HeLa acid extract probed with Histone H4 acetyl Lys12 polyclonal antibody (1:10,000 dilution).
    Lane 1: No treatment.
    Lane 2: Cells treated with sodium butyrate.

Background

Histone H4 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.

Lysine N-ε-acetylation is a dynamic, reversible and tightly regulated protein and histone modification that plays a major role in chromatin remodeling and in the regulation of gene expression in various cellular functions. The acetylation of histone H4 Lys12 is carried out by several histone acetyltransferases (HATs). Histone H4 Lys12 acetylation by TIP60 occurs in the nucleus, whereas Hat1-mediated acetylation of histone H4 Lys12 occurs in the cytoplasm, indicating the spatial regulation of histone acetylation. The acetylation of histone H4 Lys12 residue is commonly associated with transcriptional activation.

Positive Control

Active Motif's HeLa acid extract (Sodium butyrate treated) (Catalog No. 36202) can be used as a positive control for Histone H4 acetyl Lys12 antibody.

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 H4K12ac antibody (pAb).

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 H4K12ac antibody (pAb).

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