Anti-RNA polymerase II CTD repeat YSPTSPS (phospho S2) 抗体 (ab5095)

ab5095 staining RNA polymerase II CTD repeat YSPTSPS (phospho S2) in HeLa cells. The cells were fixed with 4% paraformaldehyde (10 min), permeabilized with 0.1% PBS-Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1%PBS-Tween for 1h. The cells were then incubated overnight at 4°C with ab5095 at 1µg/ml and ab7291, Mouse monoclonal [DM1A] to alpha Tubulin - Loading Control. Cells were then incubated with ab150081, Goat polyclonal Secondary Antibody to Rabbit IgG - H&L (Alexa Fluor^® 488), pre-adsorbed at 1/1000 dilution (shown in green) and ab150080, Goat polyclonal Secondary Antibody to Rabbit IgG - H&L (Alexa Fluor^® 594) at 1/1000 dilution (shown in pseudocolour red). Nuclear DNA was labelled with DAPI (shown in blue). Also suitable in cells fixed with 100% methanol (5 min).

Image was acquired with a high-content analyser (Operetta CLS, Perkin Elmer) and a maximum intensity projection of confocal sections is shown.

ab5095 staining RNA polymerase II CTD repeat YSPTSPS (phospho S2) in NIH-3T3 cells. The cells were fixed with 4% paraformaldehyde (10 min), permeabilized with 0.1% PBS-Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h. The cells were then incubated overnight at 4°C with ab5095 at 1µg/ml and ab7291, Mouse monoclonal [DM1A] to alpha Tubulin - Loading Control. Cells were then incubated with ab150081, Goat polyclonal Secondary Antibody to Rabbit IgG - H&L (Alexa Fluor^® 488), pre-adsorbed at 1/1000 dilution (shown in green) and ab150120, Goat polyclonal Secondary Antibody to Mouse IgG - H&L (Alexa Fluor^® 594), pre-adsorbed at 1/1000 dilution (shown in pseudocolour red). Nuclear DNA was labelled with DAPI (shown in blue).

Image was acquired with a confocal microscope (Leica-Microsystems TCS SP8) and a single confocal section is shown.

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. Heat mediated antigen retrieval was performed using Tris/EDTA buffer, pH9.

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.

ab12793 - S. cerevisiae RNA polymerase II CTD repeat YSPTSPS (phospho S1606 + S1613) peptide
ab18488 - Human RNA polymerase II CTD repeat YSPTSPS (phospho S5) peptide

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. 

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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. Heat mediated antigen retrieval was performed using Tris/EDTA buffer, pH9.

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.

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. Heat mediated antigen retrieval was performed using Tris/EDTA buffer, pH9.

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.

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. Heat mediated antigen retrieval was performed using Tris/EDTA buffer, pH9.

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.

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.