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Histone H2B acetyl Lys5 antibody (pAb)
| Catalog No: 39123 | Format: 200 µl | $340 | Buy Now |
| Catalog No: 39124 | Format: 10 µl | $70 | Buy Now |
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Applications Table
| Western Blot | Chromatin IP | ChIP-Seq / ChIP-chip |
|---|---|---|
| 1:2,000 - 1:10,000 | 5 - 10 µl per ChIP | modENCODE validated. |
for Histone H2B acetyl Lys5 antibody (pAb)Background
Histone H2B 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. Histone H2A and Histone H2B are acetylated in bulk chromatin by p300 and form acetylated Histone H2A/Histone H2B heterodimers. When DNA associates with intact core histone octamers that contain acetylated H2A/H2B dimers, the inhibition of transcriptional initiation significantly decreases, indicating that acetylation of their lysine residues may mediate transcription.
for Histone H2B acetyl Lys5 antibody (pAb)Immunogen
This antibody was raised against a peptide including acetyl-lysine 5 of human histone H2B.
for Histone H2B acetyl Lys5 antibody (pAb)Application Notes
This Histone H2B acetyl Lys5 antibody (H2BK5ac) has been validated for use in chromatin IP (5 - 10 µl per ChIP), ChIP-Seq & ChIP-chip (5 µl each) and Western blotting (1:2,000 - 1:10,000 dilution). Specificity for H2B acetylated at lysine 5 has been confirmed by dot blot. This Histone H2B acetyl Lys5 antibody is specific for histone H2B acetylated at lysine 5. Due to evolutionary conservation, a wide range of species reactivity is expected.
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 H2B acetyl Lys5 antibody was validated for ChIP-Seq in the study (see References).
for Histone H2B acetyl Lys5 antibody (pAb)Buffer
Rabbit serum containing 30% glycerol and 0.035% sodium azide.
for Histone H2B acetyl Lys5 antibody (pAb)Positive Control
Active Motif's HeLa acid extract (Sodium butyrate treated) (Catalog No. 36202) can be used as a positive control for Histone H2B acetyl Lys5 antibody.
Chromatin IP Assays
Active Motif has a variety of Chromatin Immunoprecipitation Kits for every need.
Please visit our Chromatin IP Page to learn about ChIP products designed for use with Histone H2B acetyl Lys5 antibody (pAb).
for Histone H2B acetyl Lys5 antibody (pAb)References
This antibody has been used as described below:
Histone H2B acetyl Lys5 pAb tested by Western blot.
HeLa acid extract (5 µg per lane) was probed with Histone H2B acetyl Lys5 polyclonal antibody (1:2,000 dilution).
Lane 1: No treatment.
Lane 2: Cells treated with sodium butyrate.
Lane 3: Human recombinant H2B (200 ng).
Histone H2B acetyl Lys5 pAb tested by dot blot analysis.
Dot blot analysis was used to confirm the specificity of Histone H2B acetyl Lys5 pAb for acetyl-Lys5 of histone H2B. Decreasing amounts of modified and unmodified peptides were spotted onto PVDF and probed with the antibody at a dilution of 1:5,000.
Lane 1: Peptide acetylated at lysine 5 of H2B.
Lane 2: Unmodified lysine 5 peptide.
Lane 3: Peptide acetylated at lysine 16 of H2B.
Lane 4: Unmodified lysine 16 peptide.
Lane 5: Peptide acetylated at lysine 120 of H2B.
Lane 6: Unmodified lysine 120 peptide.
Histone H2B acetyl Lys5 pAb tested by ChIP analysis.
ChIP performed on HeLa cell chromatin using 39123. PCR was performed using primers specific for the promoter region of the human GAPDH gene.
Lane 1: negative IgG control.
Lane 2: ChIP using 10 µl of 39123.
Lane 3: Input DNA control.
Histone H2B acetyl Lys5 pAb tested by ChIP analysis.
Chromatin IP performed using the ChIP-IT® Express Kit (Catalog No. 53008) and HeLa Chromatin (1.5 x 10^6 cell equivalents per ChIP) using 10 µl of Histone H2B acetyl Lys5 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 H2B acetyl Lys5 pAb tested by ChIP-Seq.
ChIP was performed using chromatin from the H1 human embryonic stem cell line. ChIP DNA was sequenced on the Illumina GA II and sequence tags were mapped to identify H2BK5Ac binding. The image shows H2BK5Ac binding across a 1 MB region on chromosome 1.
for Histone H2B acetyl Lys5 antibody (pAb)Storage
Antibodies in solution can be stored at -20°C for 2 years. Avoid repeated freeze/thaw cycles and keep on ice when not in storage.
for Histone H2B acetyl Lys5 antibody (pAb)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
- FACS = Flow Cytometry;
- ICC = Immunocytochemistry;
- IF = Immunofluorescence;
- IHC = Immunohistochemistry;
- IP = Immunoprecipitation;
- MeDIP = Methyl-DNA Immunoprecipitation;
- WB = Western Blotting
