Histone H4K5ac antibody (mAb)

RRID: AB_2793668
Clone: MABI 0405
Catalog No: 61523 Format: 100 µg $510 Buy

Request a quote for a bulk order Request Quote


Product Review Submit a Review


Antibody Type:
Monoclonal
Isotype:
IgG1
Purification:
Protein G Chromatography
Host:
Mouse
Molecular Weight:
8 kDa
Reactivity:
Human, Wide Range Predicted

Applications

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

Application Notes

Applications Validated by Active Motif:
ChIP: 2 - 5 µg per ChIP
WB*: 0.5 - 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:

WB
ICC/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 antibody was raised against a synthetic peptide containing acetyl-lysine 5 of human Histone H4.

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.

 

Histone H4K5ac antibody (mAb) 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 H4K5ac (mAb) or the equivalent amount of mouse 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 PABPC1 gene. Data are presented as Fold Enrichment of the ChIP antibody signal versus the negative control IgG using the ddCT method.

Histone H4K5ac antibody (mAb) tested by Western blot.
HeLa nuclear extract (20 µg per lane) probed with Histone H4K5ac (mAb) at a 2 µg/ml dilution.
Lane 1: No treatment.
Lane 2: cells treated with sodium butyrate.

Histone H4K5ac 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 H4K5ac antibody (mAb) was applied at a dilution of 0.7 µg/ml 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.

Histone H4K5ac antibody (mAb) tested by dot blot analysis.
Dot blot analysis was used to confirm the specificity of Histone H4K5ac antibody (mAb) for acetyl-Lys5 histone H4. Acetylated peptides corresponding to the immunogen and related peptides were spotted onto PVDF and probed with Histone H4K5ac antibody (mAb) at a dilution of 2 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. Lane 9: acetyl-Lys20 peptide. Lane 10: unmodified Lys20 peptide. Lane 11: acetyl-Lys31 peptide. Lane 12: unmodified Lys31 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. Histone H4 molecules acetylated at Lys5 or Lys8 are distributed in overlapping, but non-identical, islands throughout the euchromatic chromosome arms.

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 H4K5ac antibody (mAb).

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.

This antibody is manufactured by MAB Institute, Inc.

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