Astana Biomed Group, an authorized Abcam distributor in Central Asia

Anti-RNA polymerase II CTD repeat YSPTSPS (phospho S7) antibody [4E12] - BSA and Azide free (ab255847)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rat monoclonal [4E12] to RNA polymerase II CTD repeat YSPTSPS (phospho S7) - BSA and Azide free
  • Suitable for: ICC/IF, IP, WB, ChIP, ELISA, Dot blot
  • Reacts with: Mouse, Rat, Human

Overview

  • Product name

    Anti-RNA polymerase II CTD repeat YSPTSPS (phospho S7) antibody [4E12] - BSA and Azide free
    See all RNA polymerase II CTD repeat YSPTSPS primary antibodies

  • Description

    Rat monoclonal [4E12] to RNA polymerase II CTD repeat YSPTSPS (phospho S7) - BSA and Azide free

  • Host species

    Rat

  • Tested Applications & Species

    Application Species
    ChIP
    Human
    ICC/IF
    Mouse
    Human
    IP
    Human
    WB
    Mouse
    Rat
    Human
    See all applications and species data

  • Immunogen

    Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.

  • Positive control

    • WB: HeLa, RAW264.7 and PC-12 lysates. ICC/IF: HeLa and RAW 264.7 cells. IP: HeLa whole cell lysate.

  • General notes

    ab255847 is the carrier-free version of ab252853. This format is designed for use in antibody labeling, including fluorochromes, metal isotopes, oligonucleotides, enzymes.

    Our carrier-free formats are supplied in a buffer free of BSA, sodium azide and glycerol for higher conjugation efficiency.

    Use our conjugation kits for antibody conjugates that are ready-to-use in as little as 20 minutes with

    ab255847 is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm.

    Maxpar® is a trademark of Fluidigm Canada Inc.

    This product is a recombinant monoclonal antibody, which offers several advantages including:

    • - High batch-to-batch consistency and reproducibility
    • - Improved sensitivity and specificity
    • - Long-term security of supply
    • - Animal-free production
    For more information see here.

    The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing the problem with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation.

    One factor contributing to the crisis is the use of antibodies that are not suitable. This can lead to misleading results and the use of incorrect data informing project assumptions and direction. To help address this challenge, we have introduced an application and species grid on our primary antibody datasheets to make it easy to simplify identification of the right antibody for your needs.

    Learn more here.

Properties

Images

  • Immunocytochemistry/ Immunofluorescence - Anti-RNA polymerase II CTD repeat YSPTSPS (phospho S7) antibody [4E12] - BSA and Azide free (ab255847)
    Immunocytochemistry/ Immunofluorescence - Anti-RNA polymerase II CTD repeat YSPTSPS (phospho S7) antibody [4E12] - BSA and Azide free (ab255847)

    Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized HeLa cells labelling RNA polymerase II CTD repeat YSPTSPS (phospho S7) with ab252853 at 1/100 dilution, followed by ab150157 Goat Anti-Rat IgG H&L (Alexa Fluor® 488) antibody at 1/1000 dilution (Green). Confocal image showing nuclear staining in HeLa cells. ab179513 Anti-beta Tubulin rabbit monoclonal antibody was used to counterstain tubulin at 1/200 dilution (Red). The Nuclear counterstain was DAPI (Blue).

    Secondary antibody only control: Secondary antibody is ab150157 Goat Anti-Rat IgG H&L (Alexa Fluor® 488) 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 (ab252853).

  • Western blot - Anti-RNA polymerase II CTD repeat YSPTSPS (phospho S7) antibody [4E12] - BSA and Azide free (ab255847)
    Western blot - Anti-RNA polymerase II CTD repeat YSPTSPS (phospho S7) antibody [4E12] - BSA and Azide free (ab255847)
    All lanes : Anti-RNA polymerase II CTD repeat YSPTSPS (phospho S7) antibody [4E12] (ab252853) at 1/1000 dilution

    Lane 1 : PC-12 (rat adrenal gland pheochromocytoma), whole cell lysate
    Lane 2 : PC-12 whole cell lysate (Phosphatase treated membrane)

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : Goat Anti-Rat IgG H&L (HRP) (ab205720) at 1/5000 dilution

    Predicted band size: 217 kDa
    Observed band size: 260 kDa
    why is the actual band size different from the predicted?



    Blocking and diluting buffer and concentration: 5% NFDM/TBST

    The expression profile/ molecular weight  observed is consistent with what has been described in the literature (PMID: 22745433 and 23071310)

    Exposure time: 10 seconds

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab252853).

  • ChIP - Anti-RNA polymerase II CTD repeat YSPTSPS (phospho S7) antibody [4E12] - BSA and Azide free (ab255847)
    ChIP - Anti-RNA polymerase II CTD repeat YSPTSPS (phospho S7) antibody [4E12] - BSA and Azide free (ab255847)

    Chromatin was prepared from HeLa cells according to the Abcam Dual-X-ChIP protocol*. Cells were fixed with 1.5 mM EGS for 30mins and then formaldehyde for 10min. 
    The ChIP was performed with 25 µg of chromatin, 5 µg of ab252853 (red), or 5 µg of Rat IgG1 ab18407 (gray) and 20 µl of Protein A/G sepharose beads. The immunoprecipitated DNA was quantified by real time PCR (Sybr green approach). 
    http://www.abcam.com/resources?keywords=X%20ChIP%20protocol

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab252853).

  • ELISA - Anti-RNA polymerase II CTD repeat YSPTSPS (phospho S7) antibody [4E12] - BSA and Azide free (ab255847)
    ELISA - Anti-RNA polymerase II CTD repeat YSPTSPS (phospho S7) antibody [4E12] - BSA and Azide free (ab255847)

    ELISA analysis using ab252853 at a range of 1000-0 ng/ml followed by a Alkaline Phosphatase-conjugated AffiniPure Goat Anti-Rat IgG (H+L) at 1/1000 dilution. Antigen concentration: 100 ng/ml

    Antigens: phospho S2, phospho S5, phospho T4, phospho Y1, phospho S7, non-phospho

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab252853).

  • Immunoprecipitation - Anti-RNA polymerase II CTD repeat YSPTSPS (phospho S7) antibody [4E12] - BSA and Azide free (ab255847)
    Immunoprecipitation - Anti-RNA polymerase II CTD repeat YSPTSPS (phospho S7) antibody [4E12] - BSA and Azide free (ab255847)

    RNA polymerase II CTD repeat YSPTSPS (phospho S7) was immunoprecipitated from 0.35 mg HeLa (human cervix adenocarcinoma epithelial cell), whole cell lysate with ab252853 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab252853 at 1/1000 dilution. Goat Anti-Rat IgG (H+L), HRP) (ab205720) was used at 1/10000 dilution.

    Lane 1: HeLa (human cervix adenocarcinoma epithelial cell), whole cell lysate 10 ug

    Lane 2: ab252853 IP in HeLa whole cell lysate

    Lane 3: Rat monoclonal IgG1 (ab18407) instead of ab252853 in HeLa whole cell lysate

    Blocking and dilution buffer and concentration: 5% NFDM/TBST.

    Exposure time: 1 second

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab252853).

  • Immunocytochemistry/ Immunofluorescence - Anti-RNA polymerase II CTD repeat YSPTSPS (phospho S7) antibody [4E12] - BSA and Azide free (ab255847)
    Immunocytochemistry/ Immunofluorescence - Anti-RNA polymerase II CTD repeat YSPTSPS (phospho S7) antibody [4E12] - BSA and Azide free (ab255847)
    Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized RAW 264.7 cells labelling RNA polymerase II CTD repeat YSPTSPS (phospho S7) with ab252853 at 1/100 dilution, followed by ab150157 Goat Anti-Rat IgG H&L (Alexa Fluor® 488) antibody at 1/1000 2 dilution (Green). Confocal image showing nuclear staining in RAW 264.7 cells. ab179513 Anti-beta Tubulin rabbit monoclonal antibody was used to counterstain tubulin at 1/200 2.5 dilution (Red). The Nuclear counterstain was DAPI (Blue).

    Secondary antibody only control: Secondary antibody is ab150157 Goat Anti-Rat IgG H&L (Alexa Fluor® 488) at 1000 2 dilution.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab252853).

  • Western blot - Anti-RNA polymerase II CTD repeat YSPTSPS (phospho S7) antibody [4E12] - BSA and Azide free (ab255847)
    Western blot - Anti-RNA polymerase II CTD repeat YSPTSPS (phospho S7) antibody [4E12] - BSA and Azide free (ab255847)
    All lanes : Anti-RNA polymerase II CTD repeat YSPTSPS (phospho S7) antibody [4E12] (ab252853) at 1/1000 dilution

    Lane 1 : RAW264.7 (mouse Abelson murine leukemia virus-induced tumor macrophage), whole cell lysate (Untreated membrane)
    Lane 2 : RAW264.7 whole cell lysate (Phosphatase treated membrane)

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : Goat Anti-Rat IgG H&L (HRP) (ab205720) at 1/5000 dilution

    Predicted band size: 217 kDa
    Observed band size: 260 kDa why is the actual band size different from the predicted?



    Blocking and diluting buffer and concentration: 5% NFDM/TBST

    The expression profile/ molecular weight  observed is consistent with what has been described in the literature (PMID: 22745433 and 23071310) 

    Exposure time: 20 seconds

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab252853).

  • Western blot - Anti-RNA polymerase II CTD repeat YSPTSPS (phospho S7) antibody [4E12] - BSA and Azide free (ab255847)
    Western blot - Anti-RNA polymerase II CTD repeat YSPTSPS (phospho S7) antibody [4E12] - BSA and Azide free (ab255847)
    All lanes : Anti-RNA polymerase II CTD repeat YSPTSPS (phospho S7) antibody [4E12] (ab252853) at 1/1000 dilution

    Lane 1 : HeLa (human cervix adenocarcinoma epithelial cell), whole cell lysate (Untreated membrane)
    Lane 2 : HeLa whole cell lysate (Phosphatase treated membrane)

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : Goat Anti-Rat IgG H&L (HRP) (ab205720) at 1/5000 dilution

    Predicted band size: 217 kDa
    Observed band size: 260 kDa why is the actual band size different from the predicted?



    Blocking and diluting buffer and concentration: 5% NFDM/TBST

    The expression profile/ molecular weight  observed is consistent with what has been described in the literature (PMID: 22745433 and 23071310)

    Exposure time: 20 seconds

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab252853).

  • Anti-RNA polymerase II CTD repeat YSPTSPS (phospho S7) antibody [4E12] - BSA and Azide free (ab255847)
    Anti-RNA polymerase II CTD repeat YSPTSPS (phospho S7) antibody [4E12] - BSA and Azide free (ab255847)

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