Astana Biomed Group, an authorized Abcam distributor in Central Asia

Anti-p53R2 antibody [EPR8816] - BSA and Azide free (ab249090)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR8816] to p53R2 - BSA and Azide free
  • Suitable for: ICC/IF, IHC-P, WB, Flow Cyt, IP
  • Knockout validated
  • Reacts with: Human

Overview

  • Product name

    Anti-p53R2 antibody [EPR8816] - BSA and Azide free
    See all p53R2 primary antibodies

  • Description

    Rabbit monoclonal [EPR8816] to p53R2 - BSA and Azide free

  • Host species

    Rabbit

  • Tested applications

    Suitable for: ICC/IF, IHC-P, WB, Flow Cyt, IPmore details

  • Species reactivity

    Reacts with: Human

  • Immunogen

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

  • Positive control

    • WB: HeLa, HCT116, MCF7, Human skeletal muscle, and SW480 lysates. IHC-P: Human breast carcinoma tissue. ICC/IF: HeLa cells. Flow Cyt: MCF7 cells. IP: MCF7 cells.

  • General notes

    ab249090 is the carrier-free version of ab154194. 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

    ab249090 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.

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.

    We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated 'PUR' on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.

Properties

Images

  • Western blot - Anti-p53R2 antibody [EPR8816] - BSA and Azide free (ab249090)
    Western blot - Anti-p53R2 antibody [EPR8816] - BSA and Azide free (ab249090)
    All lanes : Anti-p53R2 antibody [EPR8816] (ab154194) at 1/1000 dilution

    Lane 1 : Wild-type HeLa cell lysate
    Lane 2 : RRM2B knockout HeLa cell lysate

    Lysates/proteins at 40 µg per lane.

    Performed under reducing conditions.

    Predicted band size: 40 kDa



    This data was developed using the same antibody clone in a different buffer formulation (ab154194).

    Lanes 1- 2: Merged signal (red and green). Green - ab154194 observed at 40 kDa. Red - Anti-alpha Tubulin antibody [DM1A] - Loading Control (ab7291) observed at 50 kDa.

     ab154194 was shown to react with p53R2 in wild-type HeLa cells in western blot. Loss of signal was observed when knockout cell line ab261769 (knockout cell lysate ab257215) was used. Wild-type HeLa and RRM2B knockout HeLa cell lysates were subjected to SDS-PAGE. Membrane was blocked for 1 hour at room temperature in 0.1% TBST with 3% non-fat dried milk. ab154194 and Anti-alpha Tubulin antibody [DM1A] - Loading Control (ab7291) overnight at 4°C at a 1 in 1000 dilution and a 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye®800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye®680RD) preadsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.

  • Flow Cytometry - Anti-p53R2 antibody [EPR8816] - BSA and Azide free (ab249090)
    Flow Cytometry - Anti-p53R2 antibody [EPR8816] - BSA and Azide free (ab249090)

    Flow Cytometry analysis of MCF7 (Human breast adenocarcinoma epithelial cell) cells labeling p53R2 with purified ab154194 at 1/20 dilution (10 µg/ml) (Red). Cells were fixed with 4% Paraformaldehyde and permeabilised with 90% Methanol. A Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) secondary antibody was used at 1/2000. Isotype control - Rabbit monoclonal IgG (Black). Unlabeled control - Cell without incubation with primary antibody and secondary antibody (Blue).

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab154194).

  • Immunocytochemistry/ Immunofluorescence - Anti-p53R2 antibody [EPR8816] - BSA and Azide free (ab249090)
    Immunocytochemistry/ Immunofluorescence - Anti-p53R2 antibody [EPR8816] - BSA and Azide free (ab249090)

    Immunocytochemistry/ Immunofluorescence analysis of HeLa (Human cervix adenocarcinoma epithelial cell) cells labeling p53R2 with purified ab154194 at 1/50 dilution (2.2 µg/ml). Cells were fixed in 4% Paraformaldehyde and permeabilized with 0.1% tritonX-100. Cells were counterstained with ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) 1/200 (2.5 µg/ml). Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) was used as the secondary antibody at 1/1000 (2 µg/ml) dilution. DAPI (blue) was used as nuclear counterstain. PBS instead of the primary antibody was used as the secondary antibody only control.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab154194).

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-p53R2 antibody [EPR8816] - BSA and Azide free (ab249090)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-p53R2 antibody [EPR8816] - BSA and Azide free (ab249090)

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human breast carcinoma tissue sections labeling p53R2 with purified ab154194 at 1/500 dilution (0.22 µg/ml). Heat mediated antigen retrieval using Bond™ Epitope Retrieval Solution 2 (pH 9.0). Rabbit specific IHC polymer detection kit HRP/DAB (ab209101) was used as the secondary antibody. Negative control: PBS instead of the primary antibody. Hematoxylin was used as a counterstain.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab154194).

  • Western blot - Anti-p53R2 antibody [EPR8816] - BSA and Azide free (ab249090)
    Western blot - Anti-p53R2 antibody [EPR8816] - BSA and Azide free (ab249090)
    All lanes : Anti-p53R2 antibody [EPR8816] (ab154194) at 1/1000 dilution

    Lane 1 : Wild-type HCT116 cell lysate
    Lane 2 : RRM2B knockout HCT116 cell lysate

    Lysates/proteins at 20 µg per lane.

    Performed under reducing conditions.

    Predicted band size: 40 kDa
    Observed band size: 40 kDa



    This data was developed using the same antibody clone in a different buffer formulation (ab154194).

    Lanes 1- 2: Merged signal (red and green). Green - ab154194 observed at 40 kDa. Red - Anti-alpha Tubulin antibody [DM1A] - Loading Control (ab7291) observed at 50 kDa.

     ab154194 was shown to react with p53R2 in wild-type HCT116 cells in western blot. Loss of signal was observed when knockout cell line ab266897 (knockout cell lysate ab257216) was used. Wild-type HCT116 and RRM2B knockout HCT116 cell lysates were subjected to SDS-PAGE. Membrane was blocked for 1 hour at room temperature in 0.1% TBST with 3% non-fat dried milk. ab154194 and Anti-alpha Tubulin antibody [DM1A] - Loading Control (ab7291) overnight at 4°C at a 1 in 1000 dilution and a 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye®800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye®680RD) preadsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.

  • Immunoprecipitation - Anti-p53R2 antibody [EPR8816] - BSA and Azide free (ab249090)
    Immunoprecipitation - Anti-p53R2 antibody [EPR8816] - BSA and Azide free (ab249090)

    ab154194 (purified ) at 1/20 dilution (0.5ug) immunoprecipitating p53R2 in MCF7 whole cell lysate.

    Lane 1 (input): MCF7 (Human breast adenocarcinoma epithelial cell) whole cell lysate 10ug
    Lane 2 (+): ab154194 & MCF7 whole cell lysate
    Lane 3 (-): Rabbit monoclonal IgG (ab172730) instead of ab154194 in MCF7 whole cell lysate

    For western blotting, VeriBlot for IP Detection Reagent (HRP) (ab131366) was used at 1/1000 dilution.

    Blocking and diluting buffer: 5% NFDM/TBST.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab154194).

  • Anti-p53R2 antibody [EPR8816] - BSA and Azide free (ab249090)
    Anti-p53R2 antibody [EPR8816] - BSA and Azide free (ab249090)

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