Application
ICC/IF analysis of HeLa cells using H2A.Z antibody (green) and DAPI (blue). 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 H2A.Z antibody at a dilution of 1:500.Application
ICC/IF analysis of HeLa cells using antigen peptite blocked H2A.Z antibody (green) and DAPI (blue). 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 H2A.Z antibody after incubation with 10 ng/μl of the antigen peptide.Application
ICC/IF analysis of HeLa cells using control peptide blocked H2A.Z antibody (green) and DAPI (blue). 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 H2A.Z antibody after incubation with 10 ng/μl of a control peptide containing a sequence from the central region of the H2A.Z protein.Application
WB analysis of whole cell (25 μg, lane 1) and histone extracts (15 μg, lane 2) from HeLa cells, and 1 μg of recombinant histone H2A, H2B, H3 and H4 (lane 5, 6, 7 and 8, respectively) using H2A.Z antibody at a dilution of 1:1,000 in TBS-Tween containing 5% skimmed milk. Alternatively, WB was performed on histone extracts after incubation of the antibody with 1 μg of antigen peptide (1 hour at room temperature) (lane 3) or with a control peptide containing a sequence from the central region of the H2A.Z protein (lane 4).Application
ChIP was performed with HeLa extracts and either control IgG (2 μg) or Histone H2A.Z antibody (1, 2, 5 and 10 μg per ChIP experiment). The precipitated DNA was detected by qPCR with primers targeting to positive controls c-fos and EIF2S3 promoters, and to negative controls TSH2B gene and the Sat2 satellite repeat.Application
ChIP was performed with K562 extracts and either control IgG (1 μg) or Histone H2A.Z antibody (0.2, 0.5, 1 and 2 μg per ChIP experiment). The precipitated DNA was detected by qPCR with primers targeting to positive controls c-fos and EIF2S3 promoters, and to negative controls MB gene and the Sat2 satellite repeat.Application
ELISA was performed using a serial dilution of Histone H2A.Z antibody in antigen coated wells. By plotting the absorbance against the antibody dilution, the titer of the antibody was estimated to be 1:87,500.| Product Name | Histone H2A.Z antibody - ChIP grade |
|---|---|
| Antibody Type | Primary Antibodies |
| Product description | Rabbit polyclonal antibody to H2A.ZHistones are basic nuclear proteins that are responsible for the nucleosome structure of the chromosomal fiber in eukaryotes. Nucleosomes consist of approximately 146 bp of DNA wrapped around a histone 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 encodes a replication-independent member of the histone H2A family that is distinct from other members of the family. Studies in mice have shown that this particular histone is required for embryonic development and indicate that lack of functional histone H2A leads to embryonic lethality. [provided by RefSeq, Jul 2008] |
| Immunogen | Polyclonal antibody raised in rabbit against histone variant H2A. |
| Clonality | Polyclonal |
|---|---|
| Host Species | Rabbit |
| Tested Applications | ELISAICC/IFWBChIP |
| ChIP/ChIP-seq (0.5-1 µg/IP), ELISA (1:5,000), Western blotting (1:1,000), Immunofluorescence (1:500): |
|
| Species Reactivity | Human |
| Concentration | 1mg/ml |
| Purification | Affinity purified |
| Gene Synonyms | H2A/z H2A.z H2AZ H2A histone family member Z H2AFZ |
|---|---|
| Target | H2A.Z |
Application
ICC/IF analysis of HeLa cells using H2A.Z antibody (green) and DAPI (blue). 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 H2A.Z antibody at a dilution of 1:500.
Application
ICC/IF analysis of HeLa cells using antigen peptite blocked H2A.Z antibody (green) and DAPI (blue). 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 H2A.Z antibody after incubation with 10 ng/μl of the antigen peptide.
Application
ICC/IF analysis of HeLa cells using control peptide blocked H2A.Z antibody (green) and DAPI (blue). 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 H2A.Z antibody after incubation with 10 ng/μl of a control peptide containing a sequence from the central region of the H2A.Z protein.
Application
WB analysis of whole cell (25 μg, lane 1) and histone extracts (15 μg, lane 2) from HeLa cells, and 1 μg of recombinant histone H2A, H2B, H3 and H4 (lane 5, 6, 7 and 8, respectively) using H2A.Z antibody at a dilution of 1:1,000 in TBS-Tween containing 5% skimmed milk. Alternatively, WB was performed on histone extracts after incubation of the antibody with 1 μg of antigen peptide (1 hour at room temperature) (lane 3) or with a control peptide containing a sequence from the central region of the H2A.Z protein (lane 4).
Application
ChIP was performed with HeLa extracts and either control IgG (2 μg) or Histone H2A.Z antibody (1, 2, 5 and 10 μg per ChIP experiment). The precipitated DNA was detected by qPCR with primers targeting to positive controls c-fos and EIF2S3 promoters, and to negative controls TSH2B gene and the Sat2 satellite repeat.
Application
ChIP was performed with K562 extracts and either control IgG (1 μg) or Histone H2A.Z antibody (0.2, 0.5, 1 and 2 μg per ChIP experiment). The precipitated DNA was detected by qPCR with primers targeting to positive controls c-fos and EIF2S3 promoters, and to negative controls MB gene and the Sat2 satellite repeat.
Application
ELISA was performed using a serial dilution of Histone H2A.Z antibody in antigen coated wells. By plotting the absorbance against the antibody dilution, the titer of the antibody was estimated to be 1:87,500.| Application Notes | ChIP/ChIP-seq (0.5-1 µg/IP), ELISA (1:5,000), Western blotting (1:1,000), Immunofluorescence (1:500): |
|---|
| Form | Liquid |
|---|---|
| Storage Instructions | Store at -20°C; for long storage, store at -80°C. Avoid multiple freeze-thaw cycles. |
| Storage Buffer | PBS containing 0.05% azide and 0.05% ProClin 300. |
