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Western blot
ImmunocytochemistryAntibody data
- Antibody Data
- Antigen structure
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- Validations
- Western blot [1]
- ELISA [1]
- Chromatin Immunoprecipitation [1]
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- Product number
- NBP2-54618 - Provider product page
- Provider
- Novus Biologicals
- Product name
- Rabbit Polyclonal Histone H2A.Z Antibody
- Antibody type
- Polyclonal
- Description
- Affinity purified.
- Reactivity
- Human
- Host
- Rabbit
- Isotype
- IgG
- Vial size
- 50 ug
- Storage
- Store at -20C. Avoid freeze-thaw cycles.
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Supportive validation
- Submitted by
- Novus Biologicals (provider)
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- Experimental details
- Western Blot: H2AZ Antibody [NBP2-54618] - Western blot was performed on whole cell (25 ug, lane 1) and histone extracts (15 ug, lane 2) from HeLa cells, and on 1 ug of recombinant histone H2A, H2B, H3 and H4 (lane 5, 6, 7 and 8, respectively) using the antibody against H2A.Z. The antibody was diluted 1:1,000 in TBS-Tween containing 5% skimmed milk. Alternatively, Western blot was performed on histone extracts after incubation of the antibody with 1 ug of the peptide used for immunisation of the rabbit (1 hour at room temperature) (lane 3) or with a peptide containing a sequence from the central part of the H2A.Z protein (lane 4). The position of the protein of interest is indicated on the right, the marker (in kDa) is shown on the left.
Supportive validation
- Submitted by
- Novus Biologicals (provider)
- Main image
- Experimental details
- ELISA: H2AZ Antibody [NBP2-54618] - To determine the titer of the antibody, an ELISA was performed using a serial dilution of the antibody against H2A.Z. 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 1:87,500.
Supportive validation
- Submitted by
- Novus Biologicals (provider)
- Main image
- Experimental details
- Chromatin Immunoprecipitation: H2AZ Antibody [NBP2-54618] - ChIP assays were performed using human HeLa cells, the antibody against H2A.Z and optimized PCR primer pairs for qPCR. ChIP was performed using sheared chromatin from 1,000,000 cells. A titration consisting of 1, 2, 5 and 10 ug of antibody per ChIP experiment was analyzed. IgG (2 ug/IP) was used as a negative IP control. Quantitative PCR was performed with primers specific for the promoter of the active genes c-fos and EIF2S3, used as positive controls, and for the inactive TSH2B gene and the Sat2 satellite repeat, used as negative controls.
