Histone H3 (phospho T11) polyclonal antibody
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Specification
Product Description
Rabbit polyclonal antibody raised against synthetic peptide of Histone H3 (phospho T11).
Immunogen
A synthetic peptide (conjugated with KLH) corresponding to human histone H3, phosphorylated at threonine 11.
Host
Rabbit
Reactivity
Human
Form
Liquid
Purification
Affinity purification
Recommend Usage
ELISA (1:1000)
Western Blot (1:1000)
ChIP (5 ug/CHIP)
Dot Blot (1:20000)
Immunofluorescence (1:200)
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 (2 ug/IP) was used as negative IP control. QPCR was performed with primers for the EIF4A2 and GAPDH promoters, used as positive controls, and for the coding region of the inactive MYT1 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).Western Blot (Cell lysate)
Western Blot (Cell lysate) analysis of (1) 25 ug whole cell extracts of HeLa cells, (2) 15 ug histone extracts of HeLa cells treated with colcemid, (3) 1 ug of recombinant histone H2A, (4) 1 ug of recombinant histone H2B, (5) 1 ug of recombinant histone H3, and (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 H3 (phospho T11) 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 1:25700.Dot Blot
Cross reactivity tests using the Histone H3 (phospho T11) antibody.
Dot Blot analysis was performed with peptides containing other histone phosphorylations and the unmodified Histone H3 (phospho T11). One hundred to 0.2 pmol of the respective peptides were spotted on a membrane. The antibody was used at a dilution of 1:20000. The figure shows a high specificity of the antibody for the modification of interest. -
Gene Info — HIST1H3A
Entrez GeneID
8350Protein Accession#
Q93081Gene Name
HIST1H3A
Gene Alias
H3/A, H3FA
Gene Description
histone cluster 1, H3a
Omim ID
602810Gene Ontology
HyperlinkGene Summary
Histones are basic nuclear proteins that are responsible for the nucleosome structure of the chromosomal fiber in eukaryotes. This structure consists of approximately 146 bp of DNA wrapped around a nucleosome, an octamer composed of pairs of each of the four core histones (H2A, H2B, H3, and H4). The chromatin fiber is further compacted through the interaction of a linker histone, H1, with the DNA between the nucleosomes to form higher order chromatin structures. This gene is intronless and encodes a member of the histone H3 family. Transcripts from this gene lack polyA tails; instead, they contain a palindromic termination element. This gene is found in the large histone gene cluster on chromosome 6p22-p21.3. [provided by RefSeq
Other Designations
H3 histone family, member A|histone 1, H3a
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