Product Info Summary
SKU: | CI1033 |
---|---|
Size: | 50μg/66μl |
Reactive Species: | Human, Mouse |
Host: | Rabbit |
Application: | ChIP, ChIP-seq, Dot blot, ELISA, IF, WB |
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Product info
Product Name
Anti-H4K5,8,12ac HIST1H4A Antibody
SKU/Catalog Number
CI1033
Size
50μg/66μl
Form
Liquid
Description
Boster Bio Anti-H4K5,8,12ac HIST1H4A Antibody (Catalog# CI1033). Tested in ChIP, ChIP-seq, ELISA, Dot blot, WB, IF applications. This antibody reacts with Human, Mouse.
Storage & Handling
Store at -20°C. For long-term storage, store at -80°C. Avoid multiple freeze-thaw cycles.
Cite This Product
Anti-H4K5,8,12ac HIST1H4A Antibody (Boster Biological Technology, Pleasanton CA, USA, Catalog # CI1033)
Host
Rabbit
Contents
Affinity purified polyclonal antibody in PBS containing 0.05% azide and 0.05% ProClin 300.
Clonality
Polyclonal
Immunogen
This antibody is raised in rabbit against the region of histone H4 containing the acetylated lysines 5, 8 and 12 (H4K5,8,12ac), using a KLH-conjugated synthetic peptide.
*Blocking peptide can be purchased. Costs vary based on immunogen length. Contact us for pricing.
Reactive Species
CI1033 is reactive to H4C1 in Human, Mouse
Applications
CI1033 is guaranteed for ChIP, ChIP-seq, Dot blot, ELISA, IF, WB Boster Guarantee
Observed Molecular Weight
Calculated molecular weight
11.367kDa
Background of H4C1
Histones are the main constituents of the protein part of chromosomes of eukaryotic cells. They are rich in the amino acids arginine and lysine and have been greatly conserved during evolution. Histones pack the DNA into tight masses of chromatin. Two core histones of each class H2A, H2B, H3 and H4 assemble and are wrapped by 146 base pairs of DNA to form one octameric nucleosome. Histone tails undergo numerous post-translational modifications, which either directly or indirectly alter chromatin structure to facilitate transcriptional activation or repression or other nuclear processes. In addition to the genetic code, combinations of the different histone modifications reveal the so-called “histone code”. Histone methylation and demethylation is dynamically regulated by respectively histone methyl transferases and histone demethylases. Acetylation of histone H4 is associated with active genes.
Antibody Validation
Boster validates all antibodies on WB, IHC, ICC, Immunofluorescence, and ELISA with known positive control and negative samples to ensure specificity and high affinity, including thorough antibody incubations.
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Assay dilution & Images
Assay Dilutions Recommendation
The recommendations below provide a starting point for assay optimization. The actual working concentration varies and should be decided by the user.
User needs to optimize the dilution ratio for this antibody.
Validation Images & Assay Conditions
Click image to see more details
ChIP assays were performed using human K562 cells, Anti-H4K5,8,12ac polyclonal antibody (Catalog # CI1033) and optimized PCR primer sets for qPCR. A titration of the antibody consisting of 0.2, 0.5, 1 and 2 μg per ChIP experiment was analysed. IgG (1 μg/IP) was used as negative IP control. QPCR was performed with primers for promoter of the active gene EIF4A2 and for a region 1 kb upstream of the GAPDH gene, used as positive controls, and for the inactive MYOD1 gene and the Sat2 satellite repeat region used as negative controls.
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ChIP was performed with 0.5 μg of Anti-H4K5,8,12ac polyclonal antibody (Catalog # CI1033) on sheared chromatin from 100,000 K562 cells. The IP DNA was subsequently analysed on an Illumina Genome Analyzer. Library preparation, cluster generation and sequencing were performed according to the manufacturer instructions. The 36 bp tags were aligned to the human genome using the ELAND algorithm. Figure 2 shows the signal distribution along the complete length of chromosome 2 (figure 2A) and a zoomin to a 600 kb region (figure 2B). Figure 2C and D show the enrichment in two genomic regions on chromosome 3 and 12, respectively, containing EIF4A2 and GAPDH positive controls.
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To determine the titer of the antibody, an ELISA was performed using a serial dilution of Anti-H4K5,8,12ac polyclonal antibody (Catalog # CI1033) 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 1:14,500.
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Western blot analysis of H4K5,8,12ac expression in HeLa whole cell lysates (25 μg, lane 1), histone extracts from HeLa cells (15 μg, lane 2), recombinant histone H2A (1 μg, lane 3), recombinant histone H2B (1 μg, lane 4), recombinant histone H3 (1 μg, lane 5) and recombinant histone H4 (1 μg, lane 6). H4K5,8,12ac was detected using Anti-H4K5,8,12ac polyclonal antibody (Catalog # CI1033) at 1/1000 dilution.
Click image to see more details
A Dot Blot analysis was performed to test the cross reactivity of Anti-H4K5,8,12ac polyclonal antibody (Catalog # CI1033) with peptides containing other histone modifications and the unmodified H4. 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. This figure shows a high specificity of the antibody for the modification of interest.
Click image to see more details
Immunofluorescence images stained on HeLa cells: (Figure A Left) Cells stained with anti-H4K5,8,12ac polyclonal antibody (Catalog # CI1033) at 1/500 dilution. (Figure A Middle) Nuclei stained with DAPI. (Figure A Right) Merged images of two stains from the left and middle. Figure B, C, D and E: staining of the cells with the H4K5,8,12ac antibody after incubation of the antibody with 10ng/μl of the following blocking peptides: H4K5,8,12 unmodified (figure B), H4K5,8,12ac (figure C), H2A.ZK5,7,11ac (figure D) and H4K5,8,12,16ac (figure E).
Protein Target Info & Infographic
Gene/Protein Information For H4C1 (Source: Uniprot.org, NCBI)
Gene Name
H4C1
Full Name
Histone H4
Weight
11.367kDa
Superfamily
histone H4 family
Alternative Names
Histone H4 H4C1 H4-161, H4C12, H4C13, H4C14, H4C15, H4C2, H4C3, H4C4, H4C5, H4C6, H4C8, H4C9, H4FA, HIST1H4A, H4C1 H4 clustered histone 1 histone H4|H4 histone family, member A|histone 1, H4a|histone cluster 1 H4 family member a|histone cluster 1, H4a
*If product is indicated to react with multiple species, protein info is based on the gene entry specified above in "Species".For more info on H4C1, check out the H4C1 Infographic
We have 30,000+ of these available, one for each gene! Check them out.
In this infographic, you will see the following information for H4C1: database IDs, superfamily, protein function, synonyms, molecular weight, chromosomal locations, tissues of expression, subcellular locations, post-translational modifications, and related diseases, research areas & pathways. If you want to see more information included, or would like to contribute to it and be acknowledged, please contact [email protected].
Specific Publications For Anti-H4K5,8,12ac HIST1H4A Antibody (CI1033)
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No publications found for CI1033
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