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antibody from Novus Biologicals
Targeting: H3C14
H3, H3.2, H3/m, H3F2, H3FM, HIST2H3C, MGC9629
Western blot
ImmunocytochemistryAntibody 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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- Product number
- NBP2-59279 - Provider product page
- Provider
- Novus Biologicals
- Product name
- Rabbit Polyclonal Histone H3 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 H3 [ac Lys4] Antibody [NBP2-59279] - Whole cell (25 ug, lane 1) and histone extracts (15 ug, lane 2) from HeLa cells were analysed by Western blot using the antibody against H3K4ac, diluted 1:1,000 in TBSTween containing 5% BSA. Observed molecular weight is ~15 kDa.
Supportive validation
- Submitted by
- Novus Biologicals (provider)
- Main image
- Experimental details
- ELISA: Histone H3 [ac Lys4] Antibody [NBP2-59279] - To determine the titer of the antibody, an ELISA was performed using a serial dilution of the antibody directed against H3K4ac 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 1:197,500.
Supportive validation
- Submitted by
- Novus Biologicals (provider)
- Main image
- Experimental details
- Chromatin Immunoprecipitation: Histone H3 [ac Lys4] Antibody [NBP2-59279] - ChIP assays were performed using human HeLa cells, the antibody against H3K4ac and optimized PCR primer sets for qPCR. ChIP was performed using sheared chromatin from 4 million 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 GAPDH and EIF4A2 promoters, used as positive controls, and for the HBB promoter and the Sat2 satellite repeat, 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 H3 [ac Lys4] Antibody [NBP2-59279] - A Dot Blot analysis was performed to test the cross reactivity of the antibody against H3K4ac with peptides containing other histone H3 and H4 modifications and the unmodified H3K4 sequence. One hundred to 0.2 pmol of the respective peptides were spotted on a membrane. The antibody was used at a dilution of 1:5,000. Figure shows a high specificity of the antibody for the modification of interest.
