Synthetic peptide conjugated to KLH derived from within residues 1600 - 1700 of Saccharomyces cerevisiae RNA polymerase II CTD repeat YSPTSPS, phosphorylated at S2.
(Peptide available as ab12793.)
Our Abpromise guarantee covers the use of ab5095 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
应用 | Ab评论 | 说明 |
---|---|---|
ELISA | 1/438. | |
IHC-FoFr | Use at an assay dependent concentration. | |
ChIP | Use at an assay dependent concentration. | |
ChIP/Chip | Use at an assay dependent concentration. | |
IHC-P | 1/500. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. | |
IHC-Fr | Use at an assay dependent concentration. | |
ICC/IF | Use a concentration of 1 - 5 µg/ml. | |
IHC - Wholemount | Use at an assay dependent concentration. | |
Dot blot | 1/3000. AbReview 31067 | |
WB | Use a concentration of 1 µg/ml. Detects a band of approximately 240 kDa (predicted molecular weight: 217 kDa).Can be blocked with S. cerevisiae RNA polymerase II CTD repeat YSPTSPS (phospho S1606 + S1613) peptide (ab12793). | |
CHIPseq | Use 2-0.3 µg for µg of chromatin. |
Immunohistochemical analysis of paraffin-embedded rat kidney tissue labeling RNA polymerase II CTD repeat YSPTSPS (phospho S2) with ab5095 at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Weak nuclear staining on epithelium cells and glomerulus cells of rat kidney was observed. Counter stained with Hematoxylin.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.
Perform heat mediated antigen retrieval using Tris/EDTA buffer, pH9.
ab5095 stained in MCF7 cells. Cells were fixed with 100% methanol (5 min) at room temperature and incubated with PBS containing 10% goat serum, 0.3 M glycine, 1% BSA and 0.1% triton for 1h at room temperature to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody ab5095 at 1µg/ml and ab7291 (Mouse monoclonal [DM1A] to alpha Tubulin - Loading Control) at 1/1000 dilution overnight at +4°C. The secondary antibodies were ab150120 (pseudo-colored red) and ab150081 (colored green) used at 1 µg/ml for 1 hour at room temperature. DAPI was used to stain the cell nuclei (colored blue) at a concentration of 1.43 µM for 1hour at room temperature.
Ab5095 staining RNA polymerase II CTD repeat YSPTSPS (phospho S2) in Rat Brain tissue sections by IHC-FoFr (PFA perfusion fixed frozen sections). Tissue was fixed with paraformaldehyde and samples were incubated with primary antibody (1/3000 in 0.3% PBS-T) for 18hours at 20°C. An Alexa Fluor® 488 Anti-IgG Mouse polyclonal was used as the secondary antibody at 1/1000 dilution.
Diluted ab5095 was bound to immobilised phospho- or control peptides (1 microgram per mL). The antibody was detected by goat anti-rabbit IgG (HRP) (ab97080; diluted 50000 times), and signal was developed by TMB substrate.
ab5095 staining RNA polymerase II CTD repeat YSPTSPS (phospho S2) in HeLa cells by ICC/IF (Immunocytochemistry/Immunofluorescence). Cells were fixed with paraformaldehyde and permeabilized with 0.5% Triton X100. Samples were incubated with primary antibody (1/200 in PBS) for 1 hour at 22°C. An Alexa Fluor® 488 conjugated Goat Polyclonal Anti-rabbit (1/200) was used as the secondary antibody.
Immunohistochemical analysis of paraffin-embedded mouse kidney tissue labeling RNA polymerase II CTD repeat YSPTSPS (phospho S2) with ab5095 at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Nuclear staining on epithelium cells and glomerulus cells of mouse kidney was observed. Counter stained with Hematoxylin.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.
Perform heat mediated antigen retrieval using Tris/EDTA buffer, pH9.
Immunohistochemical analysis of paraffin-embedded Human tonsil tissue labeling RNA polymerase II CTD repeat YSPTSPS (phospho S2) with ab5095 at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Nuclear staining on human tonsil was observed. Counter stained with Hematoxylin.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.
Perform heat mediated antigen retrieval using Tris/EDTA buffer, pH9.
Immunohistochemical analysis of paraffin-embedded Human pancreas tissue labeling RNA polymerase II CTD repeat YSPTSPS (phospho S2) with ab5095 at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Nuclear staining on epithelium cells and pancreas islet cells of human pancreas was observed. Counter stained with Hematoxylin.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.
Perform heat mediated antigen retrieval using Tris/EDTA buffer, pH9.
IHC - Wholemount of Caenorhabditis elegans larvae labelling RNA polymerase II CTD repeat YSPTSPS (phospho S2) with ab5095. The sample was incubated with primary antibody (1/500 in PBS + 3% BSA + 0.1% Triton X-100) for 12 hours at 4°C. ab150077, an goat anti-rabbit Alexa Fluor® 488 (1/1000), was used as the secondary antibody.
Image courtesy of Human Protein Atlas
ab5095 staining in human brain, showing staining of the Purkinje cells (in brown). Paraffin embedded brain tissue was incubated with ab5095 (1:900 dilution) for 30 mins at room temperature. Antigen retrieval was performed by heat induction in citrate buffer pH 6. ab5095 was tested in a tissue microarray (TMA) containing a wide range of normal and cancer tissues as well as a cell microarray consisting of a range of commonly used, well characterised human cell lines. Further results for this antibody can be found at www.proteinatlas.org
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