Antibody data
- Antibody Data
- Antigen structure
- References [0]
- Comments [0]
- Validations
- Western blot [2]
- Immunocytochemistry [1]
- Chromatin Immunoprecipitation [2]
- Other assay [1]
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Validation data
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- Product number
- NBP2-59236 - Provider product page
- Provider
- Novus Biologicals
- Product name
- Mouse Monoclonal Histone H4 Antibody
- Antibody type
- Monoclonal
- Description
- Protein A purified.
- Reactivity
- Human
- Host
- Mouse
- 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 [Methyl Lys20] Antibody (C.15200147) [NBP2-59236] - 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 H4K20me1 . The antibody was diluted 1:1,000 in TBS-Tween containing 5% skimmed milk. The marker (in kDa) is shown on the left, the position of the protein is indicated on the right
- Submitted by
- Novus Biologicals (provider)
- Main image
- Experimental details
- Western Blot: Histone H4 [Methyl Lys20] Antibody (C.15200147) [NBP2-59236] - Histone extracts of HeLa cells (15 ug) were analysed by Western blot using the monoclonal antibody against H4K20me1 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
- Immunocytochemistry/Immunofluorescence: Histone H4 [Methyl Lys20] Antibody (C.15200147) [NBP2-59236] - Human osteosarcoma (U2OS) cells were stained with the antibody against H4K20me1 and with DAPI. Cells were fixed with ice cold methanol for 10 min and blocked with PBS/TX-100 containing 5% normal goat serum. Top panel: cells were immunofluorescently labeled with the H4K20me1 antibody (left), diluted 1:200 in blocking solution followed by an anti- mouse antibody conjugated to Alexa488 or with DAPI (right), which specifically labels DNA. Bottom panel: staining of the cells with the H4K20me1 antibody after incubation of the antibody with blocking peptide at 5 ng/uL.
Supportive validation
- Submitted by
- Novus Biologicals (provider)
- Main image
- Experimental details
- Chromatin Immunoprecipitation: Histone H4 [Methyl Lys20] Antibody (C.15200147) [NBP2-59236] - ChIP assays were performed using human HeLa cells, the antibody against H4K20me1 and optimized PCR primer sets for qPCR. ChIP was performed with the the iDeal ChIP-seq kit, using sheared chromatin from 1 million cells. A titration of the antibody consisting of 0.5, 1, 2 and 5 ug per ChIP experiment was analysed. IgG (1 ug/IP) was used as negative IP control. QPCR was performed with primers for the coding region of the active GAPDH and ACTB genes, used as positive controls, and for the GAPDH promoter and a region located 1 kb upstream of the GAPDH promoter, 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).
- Submitted by
- Novus Biologicals (provider)
- Main image
- Experimental details
- Chromatin Immunoprecipitation: Histone H4 [Methyl Lys20] Antibody (C.15200147) [NBP2-59236] - ChIP assays were performed using human HeLa cells, the monclonal antibody against H4K20me1 and optimized PCR primer sets for qPCR. ChIP was performed using sheared chromatin from 10,000 cells. Two different quantities of antibody (3 and 12 uL per ChIP experiment) were analysed. IgG (1 ug/IP) was used as negative IP control. QPCR was performed with primers for the GAPDH promoter and for the inactive gene TSH2B. 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 [Methyl Lys20] Antibody (C.15200147) [NBP2-59236] - To check the specificity of the monoclonal antibody against H4K20me1, a Dot Blot was performed with with peptides containing different modifications or unmodified sequences of histone H4. One hundred to 0.2 pmol of peptide containing the respective histone modification were spotted on a membrane. The antibody was used at a dilution of 1:10,000. Figure shows a high specificity of the antibody for the modification of interest.