POL II phospho S5 Antibody, IgG1, Mouse, Monoclonal
Biozol Catalog Number:
ROC-200-301-V01
Supplier Catalog Number:
200-301-V01
Alternative Catalog Number:
ROC-200-301-V01
Manufacturer:
Rockland Immunochemicals
Host:
Mouse
Category:
Antikörper
Application:
ChIP, ELISA, IF, WB
Species Reactivity:
Human
Immunogen:
Anti-Pol II S5p Antibody was produced in mice by repeated immunization with the YSPTSPS repeat in the B1 subunit of RNA polymerase II phosphorylated at Ser5 of the repeated sequence.
Conjugation:
Unconjugated
Alternative Names:
DNA-directed RNA polymerase II subunit RPB1, RNA polymerase II subunit B1, DNA directed RNA polymerase II subunit A, DNA-directed RNA polymerase III largest subunit, RNA-directed RNA polymerase II subunit RPB1
0.01 M Sodium Phosphate, 0.25 M Sodium Chloride, pH 7.2
Form:
Liquid (sterile filtered)
Target:
Human
Antibody Type:
Primary Antibody
Application Dilute:
ELISA: 1:3,000, ChIP: 1 µg/ChIP, IF Microscopy: 1:500, WB: 1:1,000
Application Notes:
Anti-Pol II pS5 Antibody is tested for Chromatin Immunoprecipitation Sequencing, Chromatin Immunoprecipitation, ELISA, Immunofluorescence and Western Blots. Specific conditions for reactivity should be optimized by the end user. Expect a band approximate
Chromatin immunoprecipitation assays were performed using sheared chromatin from 1 million HeLa cells for Anti-Pol II S5p Antibody and optimized PCR primer pairs for qPCR. A titration consisting of 1, 2, 5 and 10 µg of antibody per ChIP experiment was analyzed. IgG (2 µg/IP) was used as a negative IP control. Quantitative PCR was performed with primers specific for the promoter and the coding region of the constitutively expressed GAPDH and ACTB genes, used as positive controls, and for exon 2 of the inactive myoglobin (MB) gene and the Sat2 satellite repeat, used as negative controls.
Western Blot results of Mouse anti-Pol II S5p antibody. Lane 1: 25 µg HeLa nuclear extracts. Primary antibody: Mouse anti-Pol II S5p antibody at 1:1,000. Secondary antibody: Peroxidase anti-mouse secondary antibody at 1:10,000 for 45 min at RT. Block: TBS-Tween containing 5% BLOTTO overnight at 4C. Predicted/Observed size: ~217 kDa for Mouse Pol II S5p.
Immunofluorescence Microscopy results of Mouse anti-Pol II S5p antibody. Tissue: HeLa cells. Fixation: methanol. Block: PBS/TX-100 containing 5% normal goat serum and 1% BSA. Primary antibody: Pol II S5p antibody at 1:500 for 1 hr at RT (left). Secondary antibody: anti-Mouse Alexa594 secondary antibody at 1:10,000 for 45 min at RT. Staining: Pol II S5p antibody as red fluorescent signal (left), DAPI blue (middle), merge of the two stainings (right).
Chromatin Immunoprecipitation was performed on sheared chromatin from 1 million HeLaS3 cells using 1 µg of Pol II S5p antibody. The IPd DNA was subsequently analyzed on an Illumina Genome Analyzer. The 36 bp tags were aligned to the human genome using the ELAND algorithm. Fig 4 shows the peak distribution along the complete sequence, a 150 kb region of the X-chromosome (fig 5), and in a two genomic regions surrounding the GAPDH (fig 6) and ACTB positive control genes (fig 7).
Chromatin Immunoprecipitation was performed on sheared chromatin from 1 million HeLaS3 cells using 1 µg of Pol II S5p antibody. The IPd DNA was subsequently analyzed on an Illumina Genome Analyzer. The 36 bp tags were aligned to the human genome using the ELAND algorithm. Fig 4 shows the peak distribution along the complete sequence, a 150 kb region of the X-chromosome (fig 5), and in a two genomic regions surrounding the GAPDH (fig 6) and ACTB positive control genes (fig 7).
Chromatin Immunoprecipitation was performed on sheared chromatin from 1 million HeLaS3 cells using 1 µg of Pol II S5p antibody. The IPd DNA was subsequently analysed on an Illumina Genome Analyzer. The 36 bp tags were aligned to the human genome using the ELAND algorithm. Fig 4 shows the peak distribution along the complete sequence, a 150 kb region of the X-chromosome (fig 5), and in a two genomic regions surrounding the GAPDH (fig 6) and ACTB positive control genes (fig 7).
Chromatin Immunoprecipitation was performed on sheared chromatin from 1 million HeLaS3 cells using 1 µg of Pol II S5p antibody. The IPd DNA was subsequently analyzed on an Illumina Genome Analyzer. The 36 bp tags were aligned to the human genome using the ELAND algorithm. Fig 4 shows the peak distribution along the complete sequence, a 150 kb region of the X-chromosome (fig 5), and in a two genomic regions surrounding the GAPDH (fig 6) and ACTB positive control genes (fig 7).
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