Antibody data
- Antibody Data
- Antigen structure
- References [0]
- Comments [0]
- Validations
- ELISA [1]
- Chromatin Immunoprecipitation [1]
- Other assay [1]
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Validation data
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- Product number
- NBP2-59253 - Provider product page
- Provider
- Novus Biologicals
- Product name
- Rabbit Polyclonal Histone H4 Antibody
- Antibody type
- Polyclonal
- Description
- Peptide 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
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- Novus Biologicals (provider)
- Main image
- Experimental details
- ELISA: Histone H4 [ac Lys13, ac Lys5, ac Lys8] Antibody [NBP2-59253] - To determine the titer of the antibody, an ELISA was performed using a serial dilution of the antibody directed against H4K5,8,12,16ac 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 1:21,200.
Supportive validation
- Submitted by
- Novus Biologicals (provider)
- Main image
- Experimental details
- Chromatin Immunoprecipitation: Histone H4 [ac Lys13, ac Lys5, ac Lys8] Antibody [NBP2-59253] - ChIP assays were performed using human HeLa cells, the antibody against H4K5,8,12,16ac 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 promoter of the active gene EIF4A2 and for a region 1 kb upstream of the GAPDH gene, used as positive controls, and for the inactive MYOD1 gene and the Sat2 satellite repeat region 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).
Supportive validation
- Submitted by
- Novus Biologicals (provider)
- Main image
- Experimental details
- Dot Blot: Histone H4 [ac Lys13, ac Lys5, ac Lys8] Antibody [NBP2-59253] - 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. Thr figure shows a high specificity of the antibody for the modification of interest.