Histone H4K12ac antibody (pAb)

RRID: AB_2793396
Catalog No: 39927 Format: 100 µg ¥88,000 Buy
Catalog No: 39928 Format: 10 µg ¥24,000 Buy

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

Applications

ChIP Validated Western Blot Validated Immunofluorescence Validated Dot Blot Validated Immunocytochemistry Validated

Application Notes

Applications Validated by Active Motif:
ChIP: 10 µg per ChIP
ICC/IF: 1 - 2 µg/ml dilution
WB*: 1 - 2 µg/ml 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.

Published Applications

The following applications have been published using this antibody. Unless noted above, Active Motif may not have validated the antibody for use in these applications:

ChIP-Seq
IF

View publications that use Active Motif products & services here. Enter the product number(s) to see publications & applications that use this antibody.

Immunogen

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

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. 39165) of this antibody is also available.

View our Guide to Histone Modifications and Biological Function.

 

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 3 µg 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 H4 acetyl Lys12 antibody tested by immunofluorescence.
HeLa cells stained at 2 µg/ml with Histone H4 acetyl Lys12 antibody. Top panel: Histone H4 acetyl Lys12 antibody. Middle panel: DAPI. Bottom panel: merge.

Histone H4 acetyl Lys12 antibody (pAb) tested by Western blot.
HeLa nuclear extract (20 µg per lane) probed with Histone H4 acetyl Lys12 antibody (1 µg per ml). Lane 1: no treatment. Lane 2: cells treated with sodium butyrate.

Histone H4 acetyl Lys12 antibody (pAb) tested by dot blot analysis.
Dot blot analysis was used to confirm the specificity of Histone H4 acetyl Lys12 antibody for acetyl Lys12 histone H4. Acetylated peptides corresponding to the immunogen and related peptides were spotted onto PVDF and probed with Histone H4 acetyl Lys12 antibody at a dilution of 1 µg/ml. The amount of peptide (picomoles) spotted is indicated next to each row.
Lane 1: acetyl-Lys5 peptide. Lane 2: unmodified Lys5 peptide. Lane 3: acetyl-Lys8 peptide. Lane 4: unmodified Lys8 peptide. Lane 5: acetyl-Lys12 peptide. Lane 6: unmodified Lys12 peptide. Lane 7: acetyl-Lys16 peptide. Lane 8: unmodified Lys16 peptide.

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). Histone H1 is a linker histone, present at the interface between the nucleosome core and DNA entry/exit points; it 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; they 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 nuclear extract (Catalog No. 36010) 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).

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