Anti-RNA polymerase II CTD repeat YSPTSPS (phospho S2) antibody [EPR18855-87] - BSA and Azide free

11 product images

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  ChIP - Anti-RNA polymerase II CTD repeat YSPTSPS (phospho S2) antibody [EPR18855-87] - BSA and Azide free (ab255866)

  Chromatin was prepared from HeLa (human epithelial cell line from cervix adenocarcinoma) cells according to the Abcam X-ChIP protocol. Cells were fixed with 1% formaldehyde for 10min.
  

  The ChIP was performed with 25 μg of chromatin, 5 μg of Anti-RNA polymerase II CTD repeat YSPTSPS (phospho S2) antibody [EPR18855-87] - ChIP Grade ab238146 (red), and 20 μl of Protein A/G sepharose beads. 5 μg of rabbit normal IgG was added to the beads control (gray). The immunoprecipitated DNA was quantified by real time PCR (sybr green approach).

  Primers and probes are located in the first kb of the transcribed region.

  This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-RNA polymerase II CTD repeat YSPTSPS (phospho S2) antibody [EPR18855-87] - ChIP Grade ab238146).

• 

  ChIP-sequencing - Anti-RNA polymerase II CTD repeat YSPTSPS (phospho S2) antibody [EPR18855-87] - BSA and Azide free (ab255866)

  This data was developed using the same antibody clone in a different buffer formulation (Anti-RNA polymerase II CTD repeat YSPTSPS (phospho S2) antibody [EPR18855-87] - ChIP Grade ab238146).
  Chromatin was prepared from HeLa cells. Cells were fixed with 1% formaldehyde for 10 minutes. ChIP was performed with 30 μg of chromatin and 4 μg of Anti-RNA polymerase II CTD repeat YSPTSPS (phospho S2) antibody [EPR18855-87] - ChIP Grade (Anti-RNA polymerase II CTD repeat YSPTSPS (phospho S2) antibody [EPR18855-87] - ChIP Grade ab238146). ChIP DNA was sequenced on the Illumina NextSeq 500 to a depth of 30 million reads. ChIP-Seq validation performed by Active Motif, Carlsbad, CA.
  Additional screenshots of mapped reads can be downloaded here.

• 

  Immunoprecipitation - Anti-RNA polymerase II CTD repeat YSPTSPS (phospho S2) antibody [EPR18855-87] - BSA and Azide free (ab255866)

  RNA polymerase II CTD repeat YSPTSPS (phospho S2) was immunoprecipitated from 0.35 mg of HeLa (human epithelial cell line from cervix adenocarcinoma) whole cell lysate with Anti-RNA polymerase II CTD repeat YSPTSPS (phospho S2) antibody [EPR18855-87] - ChIP Grade ab238146 at 1/30 dilution. Western blot was performed from the immunoprecipitate using Anti-RNA polymerase II CTD repeat YSPTSPS (phospho S2) antibody [EPR18855-87] - ChIP Grade ab238146 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (VeriBlot for IP Detection Reagent (HRP) ab131366), was used as secondary antibody at 1/5000 dilution.
  

  Lane 1: HeLa whole cell lysate 10 μg (Input).
  Lane 2: ab237146 IP in HeLa whole cell lysate.
  Lane 3: : Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) instead of Anti-RNA polymerase II CTD repeat YSPTSPS (phospho S2) antibody [EPR18855-87] - ChIP Grade ab238146 in HeLa whole cell lysate.

  Blocking and dilution buffer and concentration: 5% NFDM/TBST.
  Exposure time: 7 seconds.

  This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-RNA polymerase II CTD repeat YSPTSPS (phospho S2) antibody [EPR18855-87] - ChIP Grade ab238146).

  All lanes: Immunoprecipitation - Anti-RNA polymerase II CTD repeat YSPTSPS (phospho S2) antibody [EPR18855-87] - ChIP Grade (Anti-RNA polymerase II CTD repeat YSPTSPS (phospho S2) antibody [EPR18855-87] - ChIP Grade ab238146)

  Predicted band size: 217 kDa

  Observed band size: 270 kDa

• 

  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-RNA polymerase II CTD repeat YSPTSPS (phospho S2) antibody [EPR18855-87] - BSA and Azide free (ab255866)

  Immunohistochemical analysis of paraffin-embedded rat testis tissue labeling RNA polymerase II CTD repeat YSPTSPS (phospho S2) with Anti-RNA polymerase II CTD repeat YSPTSPS (phospho S2) antibody [EPR18855-87] - ChIP Grade ab238146 at 1/2000 dilution, followed by a Rabbit specific IHC polymer detection kit HRP/DAB (Rabbit specific IHC polymer detection kit HRP/DAB ab209101). Nuclear staining on rat testis without alkaline phosphatase treatment (panel A). No signal was detected when treated with alkaline phosphatase (panel B). Counter stained with hematoxylin.
  

  Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Rabbit specific IHC polymer detection kit HRP/DAB (Rabbit specific IHC polymer detection kit HRP/DAB ab209101).

  Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20 mins.

  The section was incubated with Anti-RNA polymerase II CTD repeat YSPTSPS (phospho S2) antibody [EPR18855-87] - ChIP Grade ab238146 for 30 mins at room temperature.

  The immunostaining was performed on a Leica Biosystems BOND® RX instrument.

  This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-RNA polymerase II CTD repeat YSPTSPS (phospho S2) antibody [EPR18855-87] - ChIP Grade ab238146).

• 

  Immunocytochemistry/ Immunofluorescence - Anti-RNA polymerase II CTD repeat YSPTSPS (phospho S2) antibody [EPR18855-87] - BSA and Azide free (ab255866)

  Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized RAW 264.7 (mouse macrophage cell line transformed with Abelson murine leukemia virus) cells labeling RNA polymerase II CTD repeat YSPTSPS (phospho S2) with Anti-RNA polymerase II CTD repeat YSPTSPS (phospho S2) antibody [EPR18855-87] - ChIP Grade ab238146 at 1/500 dilution, followed by a Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing nuclear staining in RAW 264.7 cell line, the signal decreased after phosphatase treatment at 37°C for 2h. The nuclear counter stain is DAPI (blue). Tubulin is detected with Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor^® 594) (Alexa Fluor® 594 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker ab195889) at 1/200 dilution (red).
  

  Secondary antibody only control: Used PBS instead of primary antibody, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077) secondary antibody at 1/1000 dilution.

  This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-RNA polymerase II CTD repeat YSPTSPS (phospho S2) antibody [EPR18855-87] - ChIP Grade ab238146).

• 

  Flow Cytometry (Intracellular) - Anti-RNA polymerase II CTD repeat YSPTSPS (phospho S2) antibody [EPR18855-87] - BSA and Azide free (ab255866)

  Intracellular flow cytometric analysis of4% paraformaldehyde-fixed, 90% methanol-permeabilized HeLa (human epithelial cell line from cervix adenocarcinoma) cells labeling RNA polymerase II CTD repeat YSPTSPS (phospho S2) with Anti-RNA polymerase II CTD repeat YSPTSPS (phospho S2) antibody [EPR18855-87] - ChIP Grade ab238146 at 1/600 (red) compared with Recombinant Rabbit IgG, monoclonal [EPR25A] - Isotype Control (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) (black) and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (blue).
  

  Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077), at 1/2000 dilution was used as the secondary antibody.

  This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-RNA polymerase II CTD repeat YSPTSPS (phospho S2) antibody [EPR18855-87] - ChIP Grade ab238146).

• 

  Dot Blot - Anti-RNA polymerase II CTD repeat YSPTSPS (phospho S2) antibody [EPR18855-87] - BSA and Azide free (ab255866)

  Dot blot analysis of RNA polymerase II CTD repeat YSPTSPS (phospho S2) labeled with Anti-RNA polymerase II CTD repeat YSPTSPS (phospho S2) antibody [EPR18855-87] - ChIP Grade ab238146 at 1/1000 dilution.
  

  Lane 1: RNA polymerase II CTD repeat YSPTSPS (phospho S2) peptide.
  Lane 2: RNA polymerase II CTD repeat YSPTSPS non-phospho peptide.
  Lane 3: RNA polymerase II CTD repeat YSPTSPS (phospho S5) peptide.

  Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/100000 dilution was used as secondary antibody.

  Blocking and dilution buffer: 5% NFDM/TBST.

  Exposure time: 3 minutes.

  This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-RNA polymerase II CTD repeat YSPTSPS (phospho S2) antibody [EPR18855-87] - ChIP Grade ab238146).

• 

  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-RNA polymerase II CTD repeat YSPTSPS (phospho S2) antibody [EPR18855-87] - BSA and Azide free (ab255866)

  Immunohistochemical analysis of paraffin-embedded mouse testis tissue labeling RNA polymerase II CTD repeat YSPTSPS (phospho S2) with Anti-RNA polymerase II CTD repeat YSPTSPS (phospho S2) antibody [EPR18855-87] - ChIP Grade ab238146 at 1/2000 dilution, followed by a Rabbit specific IHC polymer detection kit HRP/DAB (Rabbit specific IHC polymer detection kit HRP/DAB ab209101). Nuclear staining on mouse testis without alkaline phosphatase treatment (panel A). No signal was detected when treated with alkaline phosphatase (panel B). Counter stained with hematoxylin.
  

  Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Rabbit specific IHC polymer detection kit HRP/DAB (Rabbit specific IHC polymer detection kit HRP/DAB ab209101).

  Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20 mins.

  The section was incubated with Anti-RNA polymerase II CTD repeat YSPTSPS (phospho S2) antibody [EPR18855-87] - ChIP Grade ab238146 for 30 mins at room temperature.

  The immunostaining was performed on a Leica Biosystems BOND® RX instrument.

  This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-RNA polymerase II CTD repeat YSPTSPS (phospho S2) antibody [EPR18855-87] - ChIP Grade ab238146).

• 

  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-RNA polymerase II CTD repeat YSPTSPS (phospho S2) antibody [EPR18855-87] - BSA and Azide free (ab255866)

  Immunohistochemical analysis of paraffin-embedded human testis tissue labeling RNA polymerase II CTD repeat YSPTSPS (phospho S2) with Anti-RNA polymerase II CTD repeat YSPTSPS (phospho S2) antibody [EPR18855-87] - ChIP Grade ab238146 at 1/2000 dilution, followed by Rabbit specific IHC polymer detection kit HRP/DAB (Rabbit specific IHC polymer detection kit HRP/DAB ab209101). Nuclear staining on human testis without alkaline phosphatase treatment (panel A). No signal was detected when treated with alkaline phosphatase (panel B). Counter stained with hematoxylin.
  

  Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Rabbit specific IHC polymer detection kit HRP/DAB (Rabbit specific IHC polymer detection kit HRP/DAB ab209101).

  Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20 mins.

  The section was incubated with Anti-RNA polymerase II CTD repeat YSPTSPS (phospho S2) antibody [EPR18855-87] - ChIP Grade ab238146 for 30 mins at room temperature.

  The immunostaining was performed on a Leica Biosystems BOND® RX instrument.

  This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-RNA polymerase II CTD repeat YSPTSPS (phospho S2) antibody [EPR18855-87] - ChIP Grade ab238146).

• 

  Immunocytochemistry/ Immunofluorescence - Anti-RNA polymerase II CTD repeat YSPTSPS (phospho S2) antibody [EPR18855-87] - BSA and Azide free (ab255866)

  Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HeLa (human epithelial cell line from cervix adenocarcinoma) cells labeling RNA polymerase II CTD repeat YSPTSPS (phospho S2) with Anti-RNA polymerase II CTD repeat YSPTSPS (phospho S2) antibody [EPR18855-87] - ChIP Grade ab238146 at 1/500 dilution, followed by a Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing nuclear staining in HeLa cell line, the signal decreased after phosphatase treatment at 37°C for 2h. The nuclear counter stain is DAPI (blue). Tubulin is detected with Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor^® 594) (Alexa Fluor® 594 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker ab195889) at 1/200 dilution (red).
  

  Secondary antibody only control: Used PBS instead of primary antibody, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077) secondary antibody at 1/1000 dilution.

  This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-RNA polymerase II CTD repeat YSPTSPS (phospho S2) antibody [EPR18855-87] - ChIP Grade ab238146).

• 

  ChIP-sequencing - Anti-RNA polymerase II CTD repeat YSPTSPS (phospho S2) antibody [EPR18855-87] - BSA and Azide free (ab255866)

  This data was developed using the same antibody clone in a different buffer formulation (Anti-RNA polymerase II CTD repeat YSPTSPS (phospho S2) antibody [EPR18855-87] - ChIP Grade ab238146).
  Chromatin was prepared from HeLa cells. Cells were fixed with 1% formaldehyde for 10 minutes. ChIP was performed with 107 cells and 4 µg of Anti-RNA polymerase II CTD repeat YSPTSPS (phospho S2) antibody [EPR18855-87] - ChIP Grade (Anti-RNA polymerase II CTD repeat YSPTSPS (phospho S2) antibody [EPR18855-87] - ChIP Grade ab238146). ChIP DNA was sequenced on the Illumina NovaSeq 6000 to a depth of 30 million reads.
  Additional screenshots of mapped reads can be downloaded here.