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Western blot
ImmunocytochemistryAntibody data
- Antibody Data
- Antigen structure
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- Validations
- Western blot [1]
- Chromatin Immunoprecipitation [1]
- Other assay [1]
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- Product number
- NBP2-59246 - Provider product page
- Provider
- Novus Biologicals
- Product name
- Mouse Monoclonal Histone H4 Antibody
- Antibody type
- Monoclonal
- Description
- Protein A purified.
- Reactivity
- Human
- Host
- Mouse
- 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 [Dimethyl Lys20] Antibody (C.15200220) [NBP2-59246] - Histone extracts (15 ug) from HeLa cells were analyzed using the antibody against H4K20me2 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 (kDa) is shown on the left.
Supportive validation
- Submitted by
- Novus Biologicals (provider)
- Main image
- Experimental details
- Chromatin Immunoprecipitation: Histone H4 [Dimethyl Lys20] Antibody (C.15200220) [NBP2-59246] - ChIP assays were performed using HeLa cells, the monoclonal antibody against H4K20me2 and optimized PCR primer sets for qPCR. ChIP was performed with the Auto Histone ChIP-seq kit on the IP-Star automated system, using sheared chromatin from 1 million cells. Respectively 1 and 5 ug of antibody was used per ChIP experiment. IgG (1 ug/IP) was used as negative IP control. QPCR was performed with primers for the promoters of the GAPDH and EIF4A2 genes, used as negative controls, and for the Sat2 and SatA satellite repeats, used as positive controls. Figure shows the recovery, expressed as a % 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 H4 [Dimethyl Lys20] Antibody (C.15200220) [NBP2-59246] - To test the cross reactivity of the antibody against H4K20me2, analysis was performed with peptides containing other histone modifications and the unmodified H4K20. 100 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.
