Overview

  • Product name

    Anti-p53R2 + RRM2 antibody [EPR20651]
    See all p53R2+RRM2 primary antibodies
  • Description

    Rabbit monoclonal [EPR20651] to p53R2 + RRM2
  • Host species

    Rabbit
  • Tested applications

    Suitable for: ICC/IF, IHC-P, WB, Flow Cytmore details
  • Species reactivity

    Reacts with: Mouse, Rat, Human
  • Immunogen

    Recombinant fragment within Human RRM2 aa 50 to the C-terminus. The exact sequence is proprietary. (p53R2 Q7LG56)
    Database link: P31350

  • Positive control

    • WB: Human ovary cancer, human thyroid cancer, rat brain, HeLa, U-2 OS, PC-12, NIH/3T3, RAW264.7 and 4T1 whole cell lysates. IHC-P: Human colon, mouse spleen. ICC/IF: HeLa and NIH/3T3 cells. Flow cytometry: HeLa cells.
  • General notes

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

    This product is a recombinant rabbit monoclonal antibody.

Properties

Applications

Our Abpromise guarantee covers the use of ab209995 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes
ICC/IF 1/100.
IHC-P 1/2000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. IHC not recommended for rat due to non-specific staining in rat samples. Recommend 1:1000 dilution for mouse tissues.
WB 1/1000. Detects a band of approximately 41,45 kDa (predicted molecular weight: 45 kDa).
Flow Cyt 1/500.

Target

Images

  • All lanes : Anti-p53R2 + RRM2 antibody [EPR20651] (ab209995) at 1/10000 dilution

    Lane 1 : His-tagged human RRM2 recombinant protein, 10 ng
    Lane 2 : His-tagged human p53R2 recombinant protein, 10 ng

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

    Predicted band size: 45 kDa
    Observed band size: 38,39 kDa
    why is the actual band size different from the predicted?


    Exposure time: 1 second


    Blocking/Dilution buffer: 5% NFDM/TBST.

  • Immunohistochemical analysis of paraffin-embedded human colon tissue labeling RRM2/RRM2B with ab209995 at 1/2000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) Ready to use. Cytoplasmic and weak nucleus staining on epithelial cells of human colon (PMID: 23002339). Counter stained with hematoxylin.

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) Ready to use.

  • All lanes : Anti-p53R2 + RRM2 antibody [EPR20651] (ab209995) at 1/1000 dilution

    Lane 1 : Human ovary cancer at 20 µg
    Lane 2 : Human thyroid cancer at 20 µg
    Lane 3 : Rat brain at 20 µg
    Lane 4 : HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate at 10 µg
    Lane 5 : U-2 OS (Human bone osteosarcoma epithelial cell) whole cell lysate at 10 µg

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/50000 dilution (Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (ab97051))

    Predicted band size: 45 kDa
    Observed band size: 41,45 kDa why is the actual band size different from the predicted?



    Blocking: 5% NFDM/TBST.

    Exposure times: Lanes 1-3: 3 minutes; Lanes 4-5: 15 seconds.

    The MW of RRM2 and p53R2 observed is consistent with the literature (PMID 14661056; PMID 15670581; PMID 25213022).

  • All lanes : Anti-p53R2 + RRM2 antibody [EPR20651] (ab209995) at 1/1000 dilution

    Lane 1 : PC-12 (Rat adrenal gland pheochromocytoma) whole cell lysate
    Lane 2 : NIH/3T3 (Mouse embryonic fibroblast) whole cell lysate
    Lane 3 : RAW264.7 (Mouse Abelson murine leukemia virus-induced tumor macrophage) whole cell lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution (Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (ab97051))

    Predicted band size: 45 kDa
    Observed band size: 41,45 kDa why is the actual band size different from the predicted?


    Exposure time: 30 seconds


    Blocking/Dilution buffer: 5% NFDM/TBST.

    The MW of RRM2 and p53R2 observed is consistent with the literature (PMID 14661056; PMID 15670581; PMID 25213022).

  • Anti-p53R2 + RRM2 antibody [EPR20651] (ab209995) at 1/1000 dilution + 4T1 (mouse mammary gland carcinoma epithelial cell) whole cell lysate at 10 µg

    Secondary
    Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/50000 dilution (Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (ab97051))

    Predicted band size: 45 kDa
    Observed band size: 41,45 kDa why is the actual band size different from the predicted?


    Exposure time: 30 seconds


    Blocking/Dilution buffer: 5% NFDM/TBST.

    The MW of RRM2 and p53R2 observed is consistent with the literature (PMID 14661056; PMID 15670581; PMID 25213022).

  • Immunohistochemical analysis of paraffin-embedded mouse spleen tissue labeling RRM2/RRM2B with ab209995 at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) Ready to use. Cytoplasmic staining on mouse spleen (PMID: 25838540). Counter stained with hematoxylin.

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) Ready to use.

  • Immunofluorescent analysis of 100% methanol-fixed HeLa (Human cervix adenocarcinoma epithelial cell) cells labeling RRM2+ p53R2 with ab209995 at 1/100 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green).

    Confocal image showing cytoplasmic staining on HeLa cells (PMID: 194169810).

    The nuclear counterstain is DAPI (blue).

    Tubulin is detected with ab195889 (Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594)) at 1/200 dilution (red).

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) at 1/1000 dilution.

  • Immunofluorescent analysis of 100% methanol-fixed NIH/3T3 (mouse embryonic fibroblast) cells labeling RRM2+ p53R2 with ab209995 at 1/100 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green).

    Confocal image showing cytoplasmic staining on NIH/3T3 cells.

    The nuclear counterstain is DAPI (blue).

    Tubulin is detected with ab195889 (Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594)) at 1/200 dilution (red).

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) at 1/1000 dilution.

  • Flow cytometric analysis of 80% methanol fixed, 0.1% Tween-20 permeabilized HeLa (human cervix adenocarcinoma epithelial cell) cell line labeling p53R2 + RRM2 with ab209995 at 1/500 dilution (right panel) compared with a Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730) (left panel). Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) at 1/2000 dilution was used as the secondary antibody.

References

ab209995 has not yet been referenced specifically in any publications.

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