Histone H4 [ac Lys8] Antibody - BSA Free

Western Blot: Histone H4 [ac Lys8] Antibody [NBP2-59260] - Histone extracts of HeLa cells (15 ug) were analysed by Western blot using the antibody directed against H4K8ac diluted 1:200 in TBS-Tween containing 5% skimmed milk. The position of the protein of interest is indicated on the right; the marker (in kDa) is shown on the left.

Immunocytochemistry/Immunofluorescence: Histone H4 [ac Lys8] Antibody [NBP2-59260] - Mouse NIH3T3 cells were stained with the antibody against H4K8ac and with DAPI. Cells were fixed with 4% formaldehyde for 10' and blocked with PBS/TX-100 containing 5% normal goat serum and 1% BSA. The cells were immunofluorescently labelled with the H4K8ac antibody (left) diluted 1:500 in blocking solution followed by an anti-rabbit antibody conjugated to Alexa Fluor 488. The middle panel shows staining of the nuclei with DAPI. A merge of the two stainings is shown on the right.

Chromatin Immunoprecipitation: Histone H4 [ac Lys8] Antibody [NBP2-59260] - ChIP assays were performed using human HeLa cells, the antibody directed against H4K8ac and optimized PCR primer sets for qPCR. ChIP was performed with a LowCell l# ChIP kit on sheared chromatin from 10,000 cells. A titration of the antibody consisting of 1, 2, 5, and 10 ug per ChIP experiment was analysed. IgG (1 ug/IP) was used as negative IP control. QPCR was performed with primers for the c-fos promoter and for the Sat2 satellite repeat region. Figure shows the recovery, expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis).

ELISA: Histone H4 [ac Lys8] Antibody [NBP2-59260] - To determine the titer of the antibody, an ELISA was performed using a serial dilution of the antibody directed against H4K8ac, crude serum and flow through in antigen coated wells. The antigen used was a peptide containing the histone modification of interest. By plotting the absorbance against the antibody dilution, the titer of the purified antibody was estimated to be 1:16,700.

Dot Blot: Histone H4 [ac Lys8] Antibody [NBP2-59260] - A Dot Blot analysis was performed to test the cross reactivity of the antibody against H4K8ac with peptides containing other histone modifications and the unmodified H4K8. One hundred to 0.2 pmol of the respective peptides were spotted on a membrane. The antibody was used at a dilution of 1:20,000. Figure shows a high specificity of the antibody for the modification of interest.

• Reactivity: Hu, Mu
• Applications: WB, DB, ELISA, ICC/IF, ChIP, ICC/IF
• Clonality: Polyclonal
• Host: Rabbit
• Conjugate: Unconjugated
• Format: BSA Free

Histone H4 [ac Lys8] Antibody - BSA Free Summary

• Description: Novus Biologicals Rabbit Histone H4 [ac Lys8] Antibody - BSA Free (NBP2-59260) is a polyclonal antibody validated for use in WB, ELISA, ICC/IF and ChIP. All Novus Biologicals antibodies are covered by our 100% guarantee.
• Immunogen: The exact immunogen is proprietary information.
• Modification: ac Lys8
• Isotype: IgG
• Clonality: Polyclonal
• Host: Rabbit
• Gene: H4C16
• Purity: Peptide affinity purified

Applications/Dilutions

• Dilutions:
  • Chromatin Immunoprecipitation (ChIP) 1 ug
  • Dot Blot 1:20000
  • ELISA 1:100
  • Immunocytochemistry/ Immunofluorescence 1:500
  • Immunofluorescence 1:500
  • Western Blot 1:200

Packaging, Storage & Formulations

• Storage: Store at -20C. Avoid freeze-thaw cycles.
• Buffer: PBS
• Preservative: 0.05% Sodium Azide and 0.05% ProClin 300
• Purity: Peptide affinity purified

Alternate Names for Histone H4 [ac Lys8] Antibody - BSA Free

• H4
• H4/A
• H4/B
• H4/C
• H4/D
• H4/E
• H4/G
• H4/H
• H4/I
• H4/J
• H4/K
• H4/M
• H4/N
• H4/O
• H4/p
• H4F2
• H4FA
• H4FB
• H4FC
• H4FD
• H4FE
• H4FG
• H4FH
• H4FI
• H4FJ
• H4FK
• H4FM
• H4FN
• H4FO
• H4K8ac
• H4M
• HIST1H4A
• HIST1H4B
• HIST1H4C
• HIST1H4D
• HIST1H4E
• HIST1H4F
• HIST1H4H
• HIST1H4I
• HIST1H4J
• HIST1H4K
• HIST1H4L
• HIST2H4
• HIST2H4A
• HIST2H4B
• HIST4H4
• histone 4, H4
• Histone Cluster 1 H4
• Histone Cluster 1 H4i
• Histone Cluster 4 H4
• histone cluster 4, H4
• Histone H4
• MGC24116

Background

Histone proteins H3, H4, H2A, and H2B function as building blocks to package eukaryotic DNA into repeating nucleosome units that are folded in higher order chromatin fibers. The nucleosome is composed of an octamer containing a H3/H4 tetramer and two H2A/H2B dimers, surrounded by approximately 146 base pairs of DNA. A diverse and elaborate array of post-translational modifications including acetylation, phosphorylation, methylation, ubiquitination, and ADP-ribosylation occurs on the N-terminal tail domains of histones. Methylation of position-specific lysine residues in histone N termini is a central modification for regulating epigenetic transitions in chromatin. Each methylatable lysine residue can exist in a mono, di, or tri methylated state. Arginine resdiues can also by mono or di methylated.

Limitations

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

Bioinformatics

• Gene Symbol: H4C16