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Cat Number:CI1050
Size:50ug
Concentration:50ug/47ul
Price:£390.00
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Shipping:£10.00

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H4K20me3, Rabbit Polyclonal Antibody

Partner: Boster Biological Technology

Applications:ChIP, ChIP-qPCR, ChIP-seq, Flow Cytometry, IF, IHC, WB
Species Reactivity:Human, Mouse
Isotype:Rabbit IgG
Immunogen:This antibody is raised in rabbit against the region of histone H4 containing the trimethylated lysine 20 (H4K20me3), using a KLH-conjugated synthetic peptide.
Conjugation:Unconjugated
Description:Rabbit Polyclonal H4K20me3 Antibody. Validated in ChIP, ChIP-qPCR, ChIP-seq, Flow Cytometry, IF, IHC, WB and tested in Human, Mouse.
Synonym:Histone H4;HIST1H4A;H4/A, H4FA;HIST1H4B;H4/I, H4FI;HIST1H4C;H4/G, H4FG;HIST1H4D;H4/B, H4FB;HIST1H4E;H4/J, H4FJ;HIST1H4F;H4/C, H4FC;HIST1H4H;H4/H, H4FH;HIST1H4I;H4/M, H4FM;HIST1H4J;H4/E, H4FE;HIST1H4K;H4/D, H4FD;HIST1H4L;H4/K, H4FK;HIST2H4A;H4/N, H4F2, H4FN, HIST2H4;HIST2H4B;H4/O, H4FO;HIST4H4;
Storage:At -20C for one year, at 4C for one month. Avoid repeated freezing and thawing.
Usage:Research use only

 

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ChIP assays were performed using human HeLa cells, Anti-H4K20me3 polyclonal antibody (Catalog # CI1050) and optimized PCR primer sets for qPCR. A titration of the antibody consisting of 1, 2, 5, and 10 ??g per ChIP experiment was analysed. IgG (1 ??g/IP) was used as negative IP control. QPCR was performed with primers for promoters of the active genes c-fos and GAPDH, used as negative controls, and for the Sat2 satellite repeat region used as a positive control.|ChIP was performed with 1 μg of Anti-H4K20me3 polyclonal antibody (Catalog # CI1050) on sheared chromatin from 1 million HeLaS3 cells. The IP’d DNA was analysed by QPCR with optimized PCR primer pairs for the promoter and coding region of the active GAPDH gene, for the coding region of the ZNF510 gene and for the Sat2 satellite repeat (figure 2A). The IP’d DNA was subsequently analysed on an Illumina Genome Analyzer. Library preparation, cluster generation and sequencing were performed according to the manufacturer’s instructions. The 36 bp tags were aligned to the human genome using the ELAND algorithm. Figure 2B shows the signal distribution along the long arm of chromosome 19 and a zoomin to an enriched region containing several ZNF repeat genes. Figure 2C and D show the enrichment at ZNF12 and ZNF510 on chromosome 7 and 9, respectively. These results clearly show anenrichment of H4K20me3 at ZNF repeat genes.|ChIP was performed with 1 μg of Anti-H4K20me3 polyclonal antibody (Catalog # CI1050) on sheared chromatin from 1 million HeLaS3 cells. The IP’d DNA was analysed by QPCR with optimized PCR primer pairs for the promoter and coding region of the active GAPDH gene, for the coding region of the ZNF510 gene and for the Sat2 satellite repeat (figure 2A). The IP’d DNA was subsequently analysed on an Illumina Genome Analyzer. Library preparation, cluster generation and sequencing were performed according to the manufacturer’s instructions. The 36 bp tags were aligned to the human genome using the ELAND algorithm. Figure 2B shows the signal distribution along the long arm of chromosome 19 and a zoomin to an enriched region containing several ZNF repeat genes. Figure 2C and D show the enrichment at ZNF12 and ZNF510 on chromosome 7 and 9, respectively. These results clearly show anenrichment of H4K20me3 at ZNF repeat genes.|A Dot Blot analysis was performed to test the cross reactivity of Anti-H4K20me3 polyclonal antibody (Catalog # CI1050) with peptides containing other histone modifications and the unmodified H4K20. 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. This figure shows a high specificity of the antibody for the modification of interest.|To determine the titer of the antibody, an ELISA was performed using a serial dilution of Anti-H4K20me3 polyclonal antibody (Catalog # CI1050), 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 antibody was estimated to be 1:7,400.|Western blot analysis of H4K20me3 expression in histone extracts from HeLa cells (15 μg). H4K20me3 was detected using Anti-H4K20me3 polyclonal antibody (Catalog # CI1050) at 1/1000 dilution.|Immunofluorescence images stained on Human osteosarcoma (U2OS) cells: (Figure A left) Cells stained with anti-H4K20me3 polyclonal antibody (Catalog # CI1050) at 1/300. (Figure A right) Nuclei stained with DAPI. (Figure B) staining of the cells with the H4K20me3 antibody after incubation of the antibody with 5 ng/μl blocking peptide.|

ChIP assays were performed using human HeLa cells, Anti-H4K20me3 polyclonal antibody (Catalog # CI1050) and optimized PCR primer sets for qPCR. A titration of the antibody consisting of 1, 2, 5, and 10 ??g per ChIP experiment was analysed. IgG (1 ??g/IP) was used as negative IP control. QPCR was performed with primers for promoters of the active genes c-fos and GAPDH, used as negative controls, and for the Sat2 satellite repeat region used as a positive control.|ChIP was performed with 1 μg of Anti-H4K20me3 polyclonal antibody (Catalog # CI1050) on sheared chromatin from 1 million HeLaS3 cells. The IP’d DNA was analysed by QPCR with optimized PCR primer pairs for the promoter and coding region of the active GAPDH gene, for the coding region of the ZNF510 gene and for the Sat2 satellite repeat (figure 2A). The IP’d DNA was subsequently analysed on an Illumina Genome Analyzer. Library preparation, cluster generation and sequencing were performed according to the manufacturer’s instructions. The 36 bp tags were aligned to the human genome using the ELAND algorithm. Figure 2B shows the signal distribution along the long arm of chromosome 19 and a zoomin to an enriched region containing several ZNF repeat genes. Figure 2C and D show the enrichment at ZNF12 and ZNF510 on chromosome 7 and 9, respectively. These results clearly show anenrichment of H4K20me3 at ZNF repeat genes.|ChIP was performed with 1 μg of Anti-H4K20me3 polyclonal antibody (Catalog # CI1050) on sheared chromatin from 1 million HeLaS3 cells. The IP’d DNA was analysed by QPCR with optimized PCR primer pairs for the promoter and coding region of the active GAPDH gene, for the coding region of the ZNF510 gene and for the Sat2 satellite repeat (figure 2A). The IP’d DNA was subsequently analysed on an Illumina Genome Analyzer. Library preparation, cluster generation and sequencing were performed according to the manufacturer’s instructions. The 36 bp tags were aligned to the human genome using the ELAND algorithm. Figure 2B shows the signal distribution along the long arm of chromosome 19 and a zoomin to an enriched region containing several ZNF repeat genes. Figure 2C and D show the enrichment at ZNF12 and ZNF510 on chromosome 7 and 9, respectively. These results clearly show anenrichment of H4K20me3 at ZNF repeat genes.|A Dot Blot analysis was performed to test the cross reactivity of Anti-H4K20me3 polyclonal antibody (Catalog # CI1050) with peptides containing other histone modifications and the unmodified H4K20. 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. This figure shows a high specificity of the antibody for the modification of interest.|To determine the titer of the antibody, an ELISA was performed using a serial dilution of Anti-H4K20me3 polyclonal antibody (Catalog # CI1050), 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 antibody was estimated to be 1:7,400.|Western blot analysis of H4K20me3 expression in histone extracts from HeLa cells (15 μg). H4K20me3 was detected using Anti-H4K20me3 polyclonal antibody (Catalog # CI1050) at 1/1000 dilution.|Immunofluorescence images stained on Human osteosarcoma (U2OS) cells: (Figure A left) Cells stained with anti-H4K20me3 polyclonal antibody (Catalog # CI1050) at 1/300. (Figure A right) Nuclei stained with DAPI. (Figure B) staining of the cells with the H4K20me3 antibody after incubation of the antibody with 5 ng/μl blocking peptide.|