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

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-RNA polymerase II CTD repeat YSPTSPS (phospho S2) antibody (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.

• ICC/IF

AbReview24434****

Immunocytochemistry/ Immunofluorescence - Anti-RNA polymerase II CTD repeat YSPTSPS (phospho S2) antibody (AB5095)

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.

This image is courtesy of an Abreview submitted by Kirk Mcmanus.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-RNA polymerase II CTD repeat YSPTSPS (phospho S2) antibody (AB5095)

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.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-RNA polymerase II CTD repeat YSPTSPS (phospho S2) antibody (AB5095)

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.

• ICC/IF

Collaborator

Immunocytochemistry/ Immunofluorescence - Anti-RNA polymerase II CTD repeat YSPTSPS (phospho S2) antibody (AB5095)

HeLa or MCF7 cells were fixed with 4% formaldehyde in PEM buffer. The coverslip was incubated in blocking buffer of 5% powdered milk in TBS-T plus 0.02% sodium azide for 1 hour at room temperature. Blocking buffer was removed and primary antibody was added at a dilution of 1/500 and incubated overnight at 4 degrees celsius. The coverslips were then washed 4-5 times with blocking buffer for 5 minutes. Secondary antibody, goat anti-rabbit Alexa 594, was added at a dilution of 1/1000 and incubated at room temperature for one hour. From this point on coverslips were covered with foil to protect them from light. They were washed 5 times with TBS-T and then one time with PEM, for 5 minutes each wash. The coverslips were fixed 10-30 minutes in 4% formaldehyde in PEM buffer, then washed 3 times with PEM buffer for 5 minutes. 0.1M ammonium chloride in PEM buffer was added for 10 minutes to quench auto-florescence, and then slips were washed 2 times for 5 minutes in PEM followed by 3 washes for

This image is courtesy of Michael Mancini, Baylor College of Medicine

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-RNA polymerase II CTD repeat YSPTSPS (phospho S2) antibody (AB5095)

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.

Image courtesy of Human Protein Atlas

• ELISA

Lab

ELISA - Anti-RNA polymerase II CTD repeat YSPTSPS (phospho S2) antibody (AB5095)

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.

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-RNA polymerase II CTD repeat YSPTSPS (phospho S2) antibody (AB5095)

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.

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-RNA polymerase II CTD repeat YSPTSPS (phospho S2) antibody (AB5095)

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.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-RNA polymerase II CTD repeat YSPTSPS (phospho S2) antibody (AB5095)

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.

• WB

Lab

Western blot - Anti-RNA polymerase II CTD repeat YSPTSPS (phospho S2) antibody (AB5095)

All lanes:

Western blot - Anti-RNA polymerase II CTD repeat YSPTSPS (phospho S2) antibody (ab5095) at 1 µg/mL

Lanes 1, 5 and 7:

HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate at 10 µg

Lanes 2, 6 and 8:

S.cerevisiae (Y190) Whole Cell Lysate at 10 µg

Lane 3:

HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate at 10 µg with <em>S. cerevisiae</em> RNA polymerase II CTD repeat YSPTSPS peptide (<a href='/en-us/products/unavailable/s-cerevisiae-rna-polymerase-ii-ctd-repeat-ysptsps-peptide-ab12795'>ab12795</a>)

Lane 4:

S.cerevisiae (Y190) Whole Cell Lysate at 10 µg with <em>S. cerevisiae</em> RNA polymerase II CTD repeat YSPTSPS peptide (<a href='/en-us/products/unavailable/s-cerevisiae-rna-polymerase-ii-ctd-repeat-ysptsps-peptide-ab12795'>ab12795</a>)

Secondary

All lanes:

Goat polyclonal to Rabbit IgG - H&L - Pre-Adsorbed (HRP) at 1/3000 dilution

Predicted band size: 217 kDa

Observed band size: 240 kDa

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