H4K5 8 12 16ac Polyclonal Antibody
Catalogue Number: BS-53098R-BSS
BS-53098R Image
BS-53098R Image
BS-53098R Image
BS-53098R Image
bs-53098R Image
BS-53098R Image
To test the cross-reactivity of the Bioss antibody against H4K5,8,12,16ac (cat. No. bs-53098R), a Dot Blot analysis was performed 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. The figure shows a high specificity of the antibody for the modification of interest.
BS-53098R Image
To determine the titer of the antibody, an ELISA was performed using a serial dilution of the Bioss antibody directed against H4K5,8,12,16ac (cat. No. bs-53098R) 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 1:21,200.
BS-53098R Image
ChIP was performed with 2 µg of the Bioss antibody against H4K5,8,12,16ac (cat. No. bs-53098R) on sheared chromatin from 1 million HeLa cells using a ChIP-seq kit as described above. The IP'd DNA was subsequently analyzed on an Illumina Genome Analyzer. Library preparation, cluster generation, and sequencing were performed according to the manufacturer's instructions. The 36 bp tags were aligned to the human genome using the ELAND algorithm. Figure shows the signal distribution along the complete length of chromosome 5 (figure A) and a zoomin to a 600 kb region (figure B). Figure C and D show the enrichment in two genomic regions on chromosome 3 and 12, respectively, containing EIF4A2 and GAPDH positive controls. The position of the amplicon used for validating the QPCR results is shown with an arrow.
BS-53098R Image
ChIP assays were performed using human HeLa cells, the Bioss antibody against H4K5,8,12,16ac (cat. No. bs-53098R) and optimized PCR primer sets for qPCR. ChIP was performed with a ChIP- seq kit on sheared chromatin from 1 million cells. A titration of the antibody consisting of 1, 2, 5 and 10 µg per ChIP experiment was analyzed. IgG (2 µg/IP) was used as negative IP control. QPCR was performed with primers for the 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. Figure 1 shows the recovery, expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis).
bs-53098R Image
Mouse NIH3T3 cells were stained with Bioss antibody against H4K5,8,12,16ac (cat. No. bs-53098R) 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 labeled with the H4K5,8,12,16ac antibody (left) diluted 1:500 in blocking solution followed by an anti-rabbit antibody conjugated to Alexa488. The middle panel shows staining of the nuclei with DAPI. A merge of the two stainings is shown on the right.
| Manufacturer: | Bioss Inc |
| Shelf Life: | 12 months |
| Preservative: | 0.05% Sodium azide; ProClin™ 300 |
| Type: | ChIP Validated Antibodies |
| Shipping Condition: | Blue Ice |
| Unit(s): | 50 ug |
| Host name: | Rabbit |
| Clone: | |
| Isotype: | |
| Immunogen: | Polyclonal antibody raised in rabbit against the region of histone H4 containing the acetylated lysines 5, 8, 12 and 16 (H4K5,8,12,16ac), using a KLH-conjugated synthetic peptide |
| Application: | ELISA, IF, ChIP, DB |
Description
Description: Core component of nucleosome. Nucleosomes wrap and compact DNA into chromatin, limiting DNA accessibility to the cellular machineries which require DNA as a template. Histones thereby play a central role in transcription regulation, DNA repair, DNA replication and chromosomal stability. DNA accessibility is regulated via a complex set of post-translational modifications of histones, also called histone code, and nucleosome remodeling.
Additional Text
Uniprot ID
P62805
Purification
Affinity Purified
Gene ID
121504; 554313; 8294; 8359; 8360; 8361; 8362; 8363; 8364; 8365; 8366; 8367; 8368; 8370
Antibody Clonality
Polyclonal
Concentration
0.75 ug/ul
Modification
Unmodified
Gene Name
H4-16, H4C1, H4C11, H4C12, H4C13, H4C14, H4C15, H4C2, H4C3, H4C4, H4C5, H4C6, H4C8, H4C9
Storage Note
Store at -20°C for 12 months.
bs-53098R Datasheet
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