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Western blot
ImmunocytochemistryAntibody data
- Antibody Data
- Antigen structure
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- Validations
- Western blot [1]
- ELISA [1]
- Other assay [1]
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- Product number
- NBP2-59265 - 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 [Dimethyl Arg3] Antibody [NBP2-59265] - Western blot was performed on whole cell (25 ug, lane 1) and histone extracts (15 ug, lane 2) from HeLa cells, and on 1 ug of recombinant histone H2A, H2B, H3 and H4 (lane 3, 4, 5 and 6, respectively) using the antibody against H4R3me2(sym). The antibody was 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
- ELISA: Histone H4 [Dimethyl Arg3] Antibody [NBP2-59265] - To determine the titer of the antibody, an ELISA was performed using a serial dilution of the antibody directed against H4R3me2(sym) 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 1:11,750.
Supportive validation
- Submitted by
- Novus Biologicals (provider)
- Main image
- Experimental details
- Dot Blot: Histone H4 [Dimethyl Arg3] Antibody [NBP2-59265] - To test the cross reactivity of the antibody against H4R3me2(sym), a Dot Blot analysis was performed with peptides containing other histone arginine methylations and the unmodified H4R3. 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 the antibody is specific for the H4R3 symmetric dimethylation and also recognizes the H2AR3 symmetric dimethylation.
