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Western blot
ImmunocytochemistryAntibody data
- Antibody Data
- Antigen structure
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- Validations
- Western blot [1]
- ELISA [1]
- Chromatin Immunoprecipitation [2]
- Other assay [1]
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- Product number
- NBP2-54619 - Provider product page
- Provider
- Novus Biologicals
- Product name
- Rabbit Polyclonal Histone H2A.Z Antibody
- Antibody type
- Polyclonal
- Description
- Affinity purified.
- Reactivity
- Human, Mouse
- 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)
- Main image
- Experimental details
- Western Blot: Histone H2A.Z [ac Lys7, ac Lys11, ac Lys4] Antibody [NBP2-54619] - H2AZ [ac Lys4, ac Lys7, ac Lys11] Antibody [NBP2-54619] - Western blot was performed on whole cell extracts (25 ug, lane 1) from HeLa cells, and on 1 ug of recombinant histone H2A, H2B, H3 and H4 (lane 2, 3, 4 and 5, respectively) using the antibody against H2A.Zac. The antibody was diluted 1:1,000 in TBS-Tween containing 5% skimmed milk. 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: Histone H2A.Z [ac Lys7, ac Lys11, ac Lys4] Antibody [NBP2-54619] - H2AZ [ac Lys4, ac Lys7, ac Lys11] Antibody [NBP2-54619] - To determine the titer of the antibody, an ELISA was performed using a serial dilution of the antibody against H2A.Zac. The antigen used was a peptide containing the histone modifications of interest. By plotting the absorbance against the antibody dilution, the titer of the antibody was estimated to be 1:56,600.
Supportive validation
- Submitted by
- Novus Biologicals (provider)
- Main image
- Experimental details
- Chromatin Immunoprecipitation: Histone H2A.Z [ac Lys7, ac Lys11, ac Lys4] Antibody [NBP2-54619] - H2AZ [ac Lys7, ac Lys11, ac Lys4] Antibody [NBP2-54619] - H2AZ [ac Lys4, ac Lys7, ac Lys11] Antibody [NBP2-54619] - ChIP assays were performed using human HeLa cells, the antibody against H2A.Zac 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.
- Submitted by
- Novus Biologicals (provider)
- Main image
- Experimental details
- Chromatin Immunoprecipitation: Histone H2A.Z [ac Lys7, ac Lys11, ac Lys4] Antibody [NBP2-54619] - H2AZ [ac Lys7, ac Lys11, ac Lys4] Antibody [NBP2-54619] - H2AZ [ac Lys4, ac Lys7, ac Lys11] Antibody [NBP2-54619] - ChIP assays were performed using human K562 cells, the antibody against H2AZac and optimized PCR primer sets for qPCR. ChIP was performed using sheared chromatin from 100,000 cells. A titration of the antibody consisting of 0.2, 0.5, 1 and 2 ug per ChIP experiment was analysed. IgG (1 ug/IP) was used as negative IP control. Quantitative PCR was performed with primers specific for the promoter of the active genes CCT5 and EIF4A2, used as positive controls, and for the coding region of the inactive MYT1 gene and the Sat2 satellite repeat, used as negative controls. Figure shows the recovery, expressed as a percent of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis
Supportive validation
- Submitted by
- Novus Biologicals (provider)
- Main image
- Experimental details
- Dot Blot: Histone H2A.Z [ac Lys7, ac Lys11, ac Lys4] Antibody [NBP2-54619] - H2AZ [ac Lys4, ac Lys7, ac Lys11] Antibody [NBP2-54619] - To test the cross reactivity of the antibody against H2A.Zac, a Dot Blot analysis was performed with peptides containing different multiple acetylations and the unmodified H2A.Z. 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. Figure shows a high specificity of the antibody for the modification of interest.
