Astana Biomed Group, an authorized Abcam distributor in Central Asia

Anti-RNA polymerase II CTD repeat YSPTSPS (phospho S5) antibody [3E8] - BSA and Azide free (ab255846)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rat monoclonal [3E8] to RNA polymerase II CTD repeat YSPTSPS (phospho S5) - ChIP Grade – BSA and Azide free
  • Suitable for: ICC, Dot blot, ChIP, ELISA, WB
  • Reacts with: Mouse, Rat, Human

Overview

  • Product name

    Anti-RNA polymerase II CTD repeat YSPTSPS (phospho S5) antibody [3E8] - BSA and Azide free
    See all RNA polymerase II CTD repeat YSPTSPS primary antibodies

  • Description

    Rat monoclonal [3E8] to RNA polymerase II CTD repeat YSPTSPS (phospho S5) - ChIP Grade – BSA and Azide free

  • Host species

    Rat

  • Tested applications

    Suitable for: ICC, Dot blot, ChIP, ELISA, WBmore details
    Unsuitable for: IP

  • Species reactivity

    Reacts with: Mouse, Rat, Human

  • Immunogen

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

  • Positive control

    • WB: HeLa, RAW 264.7 and PC-12 whole cell lysates. ICC: HeLa and RAW264.7 cells. ChIP: Chromatin prepared from HeLa cells.

  • General notes

    ab255846 is the carrier-free version of ab252852.

    Our carrier-free antibodies are typically supplied in a PBS-only formulation, purified and free of BSA, sodium azide and glycerol. The carrier-free buffer and high concentration allow for increased conjugation efficiency.

    This conjugation-ready format is designed for use with fluorochromes, metal isotopes, oligonucleotides, and enzymes, which makes them ideal for antibody labelling, functional and cell-based assays, flow-based assays (e.g. mass cytometry) and Multiplex Imaging applications.

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

    This product is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm, without the need for antibody preparation. 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.

Properties

Images

  • ChIP - Anti-RNA polymerase II CTD repeat YSPTSPS (phospho S5) antibody [3E8] - BSA and Azide free (ab255846)
    ChIP - Anti-RNA polymerase II CTD repeat YSPTSPS (phospho S5) antibody [3E8] - BSA and Azide free (ab255846)

    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 ab252852 (red), or 5 µg of Rat IgG2a ab18450 (gray) and 20 µl of Protein A/G sepharose beads. The immunoprecipitated DNA was quantified by real time PCR (Sybr green approach).

    *https://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 (ab252852).

  • Immunocytochemistry - Anti-RNA polymerase II CTD repeat YSPTSPS (phospho S5) antibody [3E8] - BSA and Azide free (ab255846)
    Immunocytochemistry - Anti-RNA polymerase II CTD repeat YSPTSPS (phospho S5) antibody [3E8] - BSA and Azide free (ab255846)

    Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized HeLa cells labelling RNA polymerase II CTD repeat YSPTSPS (phospho S5) with ab252852 at 1/500 (1.29 μg/mL) 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 HeLa cell line, the signal decreased after phosphatase treatment at 37°C for 2h. ab179513 Anti-beta Tubulin rabbit monoclonal antibody was used to counterstain tubulin at 1/500 dilution, followed by ab150080 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 594) at 1/500 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 (ab252852).

  • Dot Blot - Anti-RNA polymerase II CTD repeat YSPTSPS (phospho S5) antibody [3E8] - BSA and Azide free (ab255846)
    Dot Blot - Anti-RNA polymerase II CTD repeat YSPTSPS (phospho S5) antibody [3E8] - BSA and Azide free (ab255846)

    RNA polymerase II CTD repeat YSPTSPS (phospho S5) labeled with ab252852 at 1/1000 (0.645 μg/mL) dilution.

    Goat Anti-Rat IgG (H+L), HRP) (ab205720) at 1/5000 dilution was used as secondary antibody.

    Lane 1: RNA polymerase II CTD repeat YSPTSPS (phospho S2) peptide

    Lane 2: RNA polymerase II CTD repeat YSPTSPS (phospho S5) peptide

    Lane 3: RNA polymerase II CTD repeat YSPTSPS (phospho T4) peptide

    Lane 4: RNA polymerase II CTD repeat YSPTSPS (phospho Y1) peptide

    Lane 5: RNA polymerase II CTD repeat YSPTSPS (phospho S7) peptide

    Lane 6: RNA polymerase II CTD repeat YSPTSPS non-phopho peptide

    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 (ab252852).

  • Western blot - Anti-RNA polymerase II CTD repeat YSPTSPS (phospho S5) antibody [3E8] - BSA and Azide free (ab255846)
    Western blot - Anti-RNA polymerase II CTD repeat YSPTSPS (phospho S5) antibody [3E8] - BSA and Azide free (ab255846)
    All lanes : Anti-RNA polymerase II CTD repeat YSPTSPS (phospho S5) antibody [3E8] - ChIP Grade (ab252852) at 0.645 µg/ml

    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: 3 seconds.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab252852).

  • ELISA - Anti-RNA polymerase II CTD repeat YSPTSPS (phospho S5) antibody [3E8] - BSA and Azide free (ab255846)
    ELISA - Anti-RNA polymerase II CTD repeat YSPTSPS (phospho S5) antibody [3E8] - BSA and Azide free (ab255846)

    ab252852 was used at 0 - 1000 ng/mL.

    Antigens were used at 1000 ng/mL.

    An Alkaline Phosphatase-conjugated Anti-Rat IgG (H+L) wasused as 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 (ab252852).

  • Western blot - Anti-RNA polymerase II CTD repeat YSPTSPS (phospho S5) antibody [3E8] - BSA and Azide free (ab255846)
    Western blot - Anti-RNA polymerase II CTD repeat YSPTSPS (phospho S5) antibody [3E8] - BSA and Azide free (ab255846)
    All lanes : Anti-RNA polymerase II CTD repeat YSPTSPS (phospho S5) antibody [3E8] - ChIP Grade (ab252852) at 0.645 µg/ml

    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: 3 seconds.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab252852).

  • Western blot - Anti-RNA polymerase II CTD repeat YSPTSPS (phospho S5) antibody [3E8] - BSA and Azide free (ab255846)
    Western blot - Anti-RNA polymerase II CTD repeat YSPTSPS (phospho S5) antibody [3E8] - BSA and Azide free (ab255846)
    All lanes : Anti-RNA polymerase II CTD repeat YSPTSPS (phospho S5) antibody [3E8] - ChIP Grade (ab252852) at 0.645 µg/ml

    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: 37 seconds.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab252852).

  • Immunocytochemistry - Anti-RNA polymerase II CTD repeat YSPTSPS (phospho S5) antibody [3E8] - BSA and Azide free (ab255846)
    Immunocytochemistry - Anti-RNA polymerase II CTD repeat YSPTSPS (phospho S5) antibody [3E8] - BSA and Azide free (ab255846)

    Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized RAW264.7 cells labelling RNA polymerase II CTD repeat YSPTSPS (phospho S5) with ab252852 at 1/500 (1.29 μg/mL) 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 RAW264.7 cell line, the signal decreased after phosphatase treatment at 37°C for 2h. ab179513 Anti-beta Tubulin rabbit monoclonal antibody was used to counterstain tubulin at 1/500 dilution, followed by ab150080 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 594) secondary antibody at 1/500 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 (ab252852).

  • Anti-RNA polymerase II CTD repeat YSPTSPS (phospho S5) antibody [3E8] - BSA and Azide free (ab255846)
    Anti-RNA polymerase II CTD repeat YSPTSPS (phospho S5) antibody [3E8] - BSA and Azide free (ab255846)

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