Explore
Validate
Learn
Western blot
ImmunocytochemistryAntibody data
- Antibody Data
- Antigen structure
- References [0]
- Comments [0]
- Validations
- Western blot [1]
- ELISA [1]
- Chromatin Immunoprecipitation [1]
- Other assay [1]
Submit
Validation data
Reference
Comment
Report error
- Product number
- NBP2-59252 - 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.
No comments: Submit comment
Supportive validation
- Submitted by
- Novus Biologicals (provider)
- Main image
- Experimental details
- Western Blot: Histone H4 [ac Lys5, ac Lys8, ac Lys12] Antibody [NBP2-59252] - 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 H4K5,8,12ac . 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 [ac Lys5, ac Lys8, ac Lys12] Antibody [NBP2-59252] - To determine the titer of the antibody, an ELISA was performed using a serial dilution of the antibody directed against H4K5,8,12ac 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 1:14,500.
Supportive validation
- Submitted by
- Novus Biologicals (provider)
- Main image
- Experimental details
- Chromatin Immunoprecipitation: Histone H4 [ac Lys5, ac Lys8, ac Lys12] Antibody [NBP2-59252] - ChIP assays were performed using human K562 cells, the antibody against H4K5,8,12ac and optimized PCR primer sets for qPCR. ChIP was performed with a ChIP- seq kit on sheared chromatin from 100,000 cells. A titration of the antibody consisting of 0.2, 0.5, 1 and 2 ug per ChIP experiment was analysed. IgG (1 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 Lys5, ac Lys8, ac Lys12] Antibody [NBP2-59252] - To test the cross reactivity of Histone H4 [ac Lys5, ac Lys8, ac Lys12] Antibody, a Dot Blot analysis was performed with peptides containing other histone modifications and the unmodified H4. 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.
