Histone H2B (K12ac) polyclonal antibody
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More Files
- More Functions
ChIP
ChIP assays were performed using human HeLa cells. A titration of the antibody consisting of 0.5, 1, 2 and, 5 ug per ChIP experiment was analysed. IgG (1 ug/IP) was used as negative IP control. QPCR was performed with primers for a region approximately 1 kb upstream of the GAPDH and ACTB promoters, used as positive controls, and for the coding region of the inactive MYOD1 gene and the Sat2 satellite repeat, used as negative controls. The figure shows the recovery, expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis).
ChIP-Seq
ChIP was performed on sheared chromatin from 1.5 million HeLaS3 cells using antibody. The figure shows the enrichment along the complete sequence and a 1 Mb region of the X-chromosome and in genomic regions of chromosome 7, surrounding the ACTB gene, and of chromosome 12, surrounding the GAPDH gene. The position of the amplicon used for ChIP-qPCR is indicated by an arrow.
Western Blot
Western Blot analysis of (1) 25 ug whole cell extracts of Hela cells, (2) 15 ug histone extracts of Hela cells, (3) 1 ug of recombinant histone H2A, (4) 1 ug of recombinant histone H2B, (5) 1 ug of recombinant histone H3, (6) 1 ug of recombinant histone H4.
Immunofluorescence
Immunofluorescent staining of Hela cell line with antibody followed by an anti-rabbit antibody conjugated to Alexa488 (left). The middle panel shows staining of the nuclei with DAPI. A merge of the two stainings (right).
Enzyme-linked Immunoabsorbent Assay
ELISA is a quantitative method used to determine the titer of the antibody using a serial dilution of antibody against Histone H2B (K12ac) 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 antibody was estimated to be 1:38200.
Dot Blot
Cross reactivity test using the Histone H2B (K12ac) antibody.
Dot Blot analysis was performed with peptides containing other histone acetylations and the unmodified H2B. One hundred to 0.2 pmol of the respective peptides were spotted on a membrane. The antibody was used at a dilution of 1:5000. The figure shows a high specificity of the antibody for the modification of interest.
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Specification
Product Description
Rabbit polyclonal antibody raised against synthetic peptide of Histone H2B (K12ac).
Immunogen
A synthetic peptide (conjugated with KLH) corresponding to Histone H2B, acetylated at lysine 12.
Host
Rabbit
Reactivity
Human
Form
Liquid
Purification
Affinity purification
Recommend Usage
ELISA (1:1000)
Western Blot (1:1000)
ChIP (0.5-1 ul/ChIP)
Dot Blot (1:5000)
Immunofluorescence (1:500)
The optimal working dilution should be determined by the end user.Storage Buffer
In PBS (0.05% sodium azide, 0.05% proclin 300).
Storage Instruction
Store at -20°C. For long term storage store at -80°C.
Aliquot to avoid repeated freezing and thawing.Note
This product contains sodium azide: a POISONOUS AND HAZARDOUS SUBSTANCE which should be handled by trained staff only.
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Applications
ChIP
ChIP assays were performed using human HeLa cells. A titration of the antibody consisting of 0.5, 1, 2 and, 5 ug per ChIP experiment was analysed. IgG (1 ug/IP) was used as negative IP control. QPCR was performed with primers for a region approximately 1 kb upstream of the GAPDH and ACTB promoters, used as positive controls, and for the coding region of the inactive MYOD1 gene and the Sat2 satellite repeat, used as negative controls. The figure shows the recovery, expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis).ChIP-Seq
ChIP was performed on sheared chromatin from 1.5 million HeLaS3 cells using antibody. The figure shows the enrichment along the complete sequence and a 1 Mb region of the X-chromosome and in genomic regions of chromosome 7, surrounding the ACTB gene, and of chromosome 12, surrounding the GAPDH gene. The position of the amplicon used for ChIP-qPCR is indicated by an arrow.Western Blot
Western Blot analysis of (1) 25 ug whole cell extracts of Hela cells, (2) 15 ug histone extracts of Hela cells, (3) 1 ug of recombinant histone H2A, (4) 1 ug of recombinant histone H2B, (5) 1 ug of recombinant histone H3, (6) 1 ug of recombinant histone H4.Immunofluorescence
Immunofluorescent staining of Hela cell line with antibody followed by an anti-rabbit antibody conjugated to Alexa488 (left). The middle panel shows staining of the nuclei with DAPI. A merge of the two stainings (right).Enzyme-linked Immunoabsorbent Assay
ELISA is a quantitative method used to determine the titer of the antibody using a serial dilution of antibody against Histone H2B (K12ac) 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 antibody was estimated to be 1:38200.Dot Blot
Cross reactivity test using the Histone H2B (K12ac) antibody.
Dot Blot analysis was performed with peptides containing other histone acetylations and the unmodified H2B. One hundred to 0.2 pmol of the respective peptides were spotted on a membrane. The antibody was used at a dilution of 1:5000. The figure shows a high specificity of the antibody for the modification of interest. -
Gene Info — HIST1H2BC
Entrez GeneID
8347Protein Accession#
P62807Gene Name
HIST1H2BC
Gene Alias
H2B.1, H2B/l, H2BFL, MGC104246, dJ221C16.3
Gene Description
histone cluster 1, H2bc
Omim ID
602847Gene Ontology
HyperlinkGene Summary
Histones are basic nuclear proteins that are responsible for the nucleosome structure of the chromosomal fiber in eukaryotes. Two molecules of each of the four core histones (H2A, H2B, H3, and H4) form an octamer, around which approximately 146 bp of DNA is wrapped in repeating units, called nucleosomes. The linker histone, H1, interacts with linker DNA between nucleosomes and functions in the compaction of chromatin into higher order structures. This gene is intronless and encodes a member of the histone H2B family. Transcripts from this gene lack polyA tails but instead contain a palindromic termination element. This gene is found in the large histone gene cluster on chromosome 6. [provided by RefSeq
Other Designations
H2B histone family, member L|OTTHUMP00000016141|histone 1, H2bc
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