Antibody data
- Antibody Data
- Antigen structure
- References [0]
- Comments [0]
- Validations
- Western blot [1]
- ELISA [1]
- Chromatin Immunoprecipitation [1]
- Other assay [1]
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Validation data
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- Product number
- NBP2-59260 - 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)
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- Experimental details
- Western Blot: Histone H4 [ac Lys8] Antibody [NBP2-59260] - Histone extracts of HeLa cells (15 ug) were analysed by Western blot using the antibody directed against H4K8ac diluted 1:200 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 H4 [ac Lys8] Antibody [NBP2-59260] - To determine the titer of the antibody, an ELISA was performed using a serial dilution of the antibody directed against H4K8ac, crude serum and flow through 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 purified antibody was estimated to be 1:16,700.
Supportive validation
- Submitted by
- Novus Biologicals (provider)
- Main image
- Experimental details
- Chromatin Immunoprecipitation: Histone H4 [ac Lys8] Antibody [NBP2-59260] - ChIP assays were performed using human HeLa cells, the antibody directed against H4K8ac and optimized PCR primer sets for qPCR. ChIP was performed with a LowCell l# ChIP kit on sheared chromatin from 10,000 cells. A titration of the antibody consisting of 1, 2, 5, and 10 ug per ChIP experiment was analysed. IgG (1 ug/IP) was used as negative IP control. QPCR was performed with primers for the c-fos promoter and for the Sat2 satellite repeat region. 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 Lys8] Antibody [NBP2-59260] - A Dot Blot analysis was performed to test the cross reactivity of the antibody against H4K8ac with peptides containing other histone modifications and the unmodified H4K8. 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.