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Western blotAntibody data
- Antibody Data
- Antigen structure
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- Validations
- Western blot [1]
- ELISA [1]
- Immunocytochemistry [1]
- Other assay [2]
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- Product number
- TA347269 - Provider product page
- Provider
- OriGene
- Product name
- Mouse Monoclonal Pol II S5p Antibody
- Antibody type
- Monoclonal
- Description
- Mouse Monoclonal Pol II S5p Antibody
- Host
- Mouse
- Conjugate
- Unconjugated
- Epitope
- POLR2A
- Isotype
- IgG
- Antibody clone number
- NULL
- Vial size
- 50 µg
- Concentration
- 1 ?g/?l
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Supportive validation
- Submitted by
- OriGene (provider)
- Main image
- Experimental details
- WB using the antibody against Pol II S5p 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 (in kDa) is shown on the left.
- Validation comment
- WB
Supportive validation
- Submitted by
- OriGene (provider)
- Main image
- Experimental details
- Cross reactivity of the antibody against Pol IIS5p To test the specificity an ELISA was performed using a serial dilution of the antibody against Pol IIS5p . The wells were coated with peptides containing the unmodified C-terminal repeat sequence as well as different phosphorylated peptides. Image shows the specificity of the antibody for the S5 phosphorylation.
- Validation comment
- ELISA
Supportive validation
- Submitted by
- OriGene (provider)
- Main image
- Experimental details
- HeLa cells were stained with the antibody against Pol II S5p and with DAPI. Cells were fixed with methanol and blocked with PBS/TX-100 containing 5% normal goat serum and 1% BSA. The cells were immunofluorescently labelled with the Pol II S5p antibody (left) diluted 1:500 in blocking solution followed by an anti-mouse antibody conjugated to Alexa594. The middle panel shows staining of the nuclei with DAPI. A merge of the two stainings is shown on the right.
- Validation comment
- IF
Supportive validation
- Submitted by
- OriGene (provider)
- Main image
- Experimental details
- ChIP was performed on sheared chromatin from 1 million HeLaS3 cells using 1 ug of the ab against Pol II S5p. The IP'd DNA was subsequently analysed on an Illumina Genome Analyzer. The 36 bp tags were aligned to the human genome using the ELAND algorithm. Image shows the peak distribution along the complete sequence and a 150 kb region of the X-chromosome (figure 2A and B, respectively), and in a two genomic regions surrounding the GAPDH and ACTB positive control genes (figure 2C and D).
- Validation comment
- Assay
- Submitted by
- OriGene (provider)
- Main image
- Experimental details
- ChIP assays using HeLa cells: ChIP-seq kit, using sheared chromatin from 1 million cells. A titration of 1, 2, 5 and 10 ug ab was used. IgG (2ug/IP) was negative control. qPCR primers were specific for the promoter and the coding region of the constitutively expressed GAPDH and ACTB genesas positive controls, and for exon 2 of the inactive myoglobin (MB) gene and the Sat2 satellite repeatas negative controls. Image shows the recovery, expressed as a % of input (the relative amount of IP'd DNA compared to input DNA after qPCR).
- Validation comment
- Assay
