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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
- NBP2-59258 - Provider product page
- Provider
- Novus Biologicals
- Product name
- Rabbit Polyclonal Histone H4 Antibody
- Antibody type
- Polyclonal
- Description
- Peptide 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)
- Main image
- Experimental details
- Western Blot: Histone H4 [Methyl Lys20] Antibody [NBP2-59258] - 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 H4K20me1. The antibody was diluted 1:1,000 in TBS-Tween containing 5% skimmed milk. The marker (in kDa) is shown on the left, the position of the protein is indicated on the right.
Supportive validation
- Submitted by
- Novus Biologicals (provider)
- Main image
- Experimental details
- ELISA: Histone H4 [Methyl Lys20] Antibody [NBP2-59258] - To determine the titer, an ELISA was performed using a serial dilution of the antibody directed against H4K20me1 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 1:51,100.
Supportive validation
- Submitted by
- Novus Biologicals (provider)
- Main image
- Experimental details
- Chromatin Immunoprecipitation: Histone H4 [Methyl Lys20] Antibody [NBP2-59258] - ChIP assays were performed using human HeLa cells, the antibody against H4K20me1 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 ug per ChIP experiment was analysed. IgG (2 ug/IP) was used as negative IP control. QPCR was performed with primers for the coding region of the active GAPDH and ACTB genes, used as positive controls, and for the GAPDH promoter and a region located 1 kb upstream of the GAPDH promoter, used as negative controls. Figure shows the recovery, expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis).
