Histone H4K16ac antibody (pAb)

RRID: AB_2636968
Catalog No: 39167 Format: 100 µl ¥88,000 Buy
Catalog No: 39068 Format: 50 µl ¥52,000 Buy
Catalog No: 39168 Format: 10 µl ¥24,000 Buy

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

Applications

CUT&Tag Validated ChIP Validated ChIP-Seq Validated Western Blot Validated Immunofluorescence Validated Dot Blot Validated

Application Notes

Applications Validated by Active Motif:
ChIP: 10 µl per ChIP
ChIP-Seq: 10 µl each
WB: 1:1,000 - 1:5,000 dilution
CUT&Tag: 1-2 µl per 50 µl reaction

ChIP-Seq validation was performed by Active Motif's Epigenetics Services; the complete data set is available in the UCSC Genome Browser by clicking here.

For optimal results in Western blotting, primary antibody incubations should be performed overnight at 4°C. Individual optimization may be required.

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-qPCR, Chip-Seq
Native ChIP
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 Histone H4 acetyl Lys16 antibody was raised against a peptide including acetyl-lysine 16 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. 39929) of this antibody that was purified by Protein A Chromatography is also available.

 

Histone H4K16ac antibody (pAb) tested by ChIP-Seq.
ChIP was performed using the ChIP-IT® High Sensitivity Kit (Cat. No. 53040) with 15 ug of chromatin from HeLa cells and 10 µl of antibody. ChIP DNA was sequenced on the Illumina HiSeq and 12 million sequence tags were mapped to identify Histone H4K16ac binding sites. The image shows binding across a region of chromosome 5. You can view the complete data set in the UCSC Genome Browser, starting at this specific location, here.

Histone H4K16ac antibody (pAb) tested by CUT&Tag
CUT&Tag was performed using 100,000 K562 cells and sequenced using 38 base-pair, paired-end reads on the Illumina NovaSeq. Data was collected from 6 million reads, and H4K16ac data is shown for Chromosome 6.

Histone H4K16ac antibody (pAb) tested by Western blot.
HeLa nuclear extract (20 µg) probed with Histone H4 acetyl Lys16 polyclonal antibody (1:1,000 dilution).
    Lane 1: No treatment.
    Lane 2: Cells treated with sodium butyrate.

Histone H4K16ac antibody (pAb) specificity tested by peptide array analysis.
Peptide array analysis was used to confirm the specificity of this antibody for its intended modification. Histone H4 acetyl Lys16 antibody was applied at a dilution of 1:10,000 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. In the array, the acetyl Lys16 modification is found on peptides with two different backbone sequences (H4 amino acids 1-19 and amino acids 11-30) so reactive and related peptides for both peptides is shown.
Array Data File

Histone H4K16ac antibody (pAb) tested by dot blot analysis.
Dot blot analysis was used to confirm the specificity of Histone H4 acetyl Lys16 pAb for histone H4 acetyl Lys16. Acetylated peptides corresponding to the immunogen and related H4 sequences were spotted onto PVDF and probed with the antibody at 1:4,000. The amount of peptide (picomoles) spotted is indicated next to each row.
Lane 1: Unmodified Lys8 peptide. Lane 2: Acetyl Lys5 peptide. Lane 3: Acetyl Lys8 peptide. Lane 4: Acetyl Lys12 peptide. Lane 5: Unmodified Lys16 peptide. Lane 6: Acetyl Lys16 peptide. Lane 7: Acetyl Lys20 peptide. Lane 8: Unmodified Lys31 peptide. Lane 9: Acetyl Lys31 peptide. Lane 10: Acetyl Lys59 peptide. Lane 11: Acetyl Lys91 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. NoRC is a SMARCA5 (SNF2h)-containing chromatin remodeling complex. The bromodomain of TIP5, the large subunit of NoRC, interacts with acetylated Histone H4 Lys16, (H4K16ac) and cooperates with an adjacent PHD finger to recruit histone deacetylases and DNA methyltransferases to rDNA, leading to the silencing of rDNA.

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 Lys16 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 H4K16ac 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