Astana Biomed Group, an authorized Abcam distributor in Central Asia

Anti-p21 antibody [EPR3993] - BSA and Azide free (ab215971)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR3993] to p21 - BSA and Azide free
  • Suitable for: WB
  • Knockout validated
  • Reacts with: Rat, Human

Overview

  • Product name

    Anti-p21 antibody [EPR3993] - BSA and Azide free
    See all p21 primary antibodies

  • Description

    Rabbit monoclonal [EPR3993] to p21 - BSA and Azide free

  • Host species

    Rabbit

  • Specificity

    Expression levels of the target protein vary between different tissue/cell lines and in some cases induction may be required before a signal is observed.

  • Tested Applications & Species

    Application Species
    WB
    Human
    See all applications and species data

  • Immunogen

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

  • Positive control

    • WB: HUVEC, U87-MG, MCF-7, LnCaP, RAW264.7, PC-12, HCT116 and C6 cell lysates.

  • General notes

    Ab215971 is the carrier-free version of ab109199. 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

    ab215971 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.

Properties

Images

  • Western blot - Anti-p21 antibody [EPR3993] - BSA and Azide free (ab215971)
    Western blot - Anti-p21 antibody [EPR3993] - BSA and Azide free (ab215971)

    This data was developed using ab109199, the same antibody clone in a different buffer formulation.

    Lane 1 Wild-type DLD-1 cell lysate (20 µg)

    Lane 2 Wild-type DLD-1 20 µM 2,3-DCPE for 16hrs treated cell lysate (20 µg)

    Lane 3 p21 knockout DLD-1 cell lysate (20 µg)

    Lane 4 p21 knockout 20 µM 2,3-DCPE for 16hrs DLD-1 cell lysate (20 µg)

    Lane 5: HT1080 cell lysate (20 µg)

    Lanes 1 - 5 Merged signal (red and green). Green - ab109199 observed at 20 kDa. Red - loading control, ab8245, observed at 37 kDa.

    ab109199 was shown to recognize p21 in WT DLD-1 cells with 2,3-DCPE treatment along with additional cross-reactive bands. When p21 knockout DLD-1 cells +/- 2,3-DCPE treatment were used, no band was observed. Wild-type and p21 knockout samples were subjected to SDS-PAGE. ab109199 and ab8245 (loading control to GAPDH) were diluted 1/1000 and 1/10 000 respectively and incubated overnight at 4°C. 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 1/10 000 dilution for 1 h at room temperature before imaging.

  • Western blot - Anti-p21 antibody [EPR3993] - BSA and Azide free (ab215971)
    Western blot - Anti-p21 antibody [EPR3993] - BSA and Azide free (ab215971)
    All lanes : Anti-p21 antibody [EPR3993] (ab109199) at 1/1000 dilution

    Lane 1 : Wild-type HCT116 cell lysate
    Lane 2 : CDKN1A knockout HCT116 cell lysate

    Lysates/proteins at 20 µg per lane.

    Performed under reducing conditions.

    Predicted band size: 18 kDa
    Observed band size: 20 kDa why is the actual band size different from the predicted?



    This data was developed using the same antibody clone in a different buffer formulation (ab109199).

    Lanes 1- 2: Merged signal (red and green). Green - ab109199 observed at 20 kDa. Red - Anti-GAPDH antibody [6C5] - Loading Control (ab8245) observed at 37 kDa.

     ab109199 was shown to react with p21 in wild-type HCT116 cells in western blot. Loss of signal was observed when knockout cell line ab266860 (knockout cell lysate ab256870) was used. Wild-type HCT116 and CDKN1A 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. ab109199 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) 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.

  • Western blot - Anti-p21 antibody [EPR3993] - BSA and Azide free (ab215971)
    Western blot - Anti-p21 antibody [EPR3993] - BSA and Azide free (ab215971)
    Lane 1 : Anti-p21 antibody [EPR3993] (ab109199) (0.7ug/ul)
    Lane 2 : Anti-p21 antibody [EPR362] (ab109520) (0.7ug/ul)

    All lanes : MCF-7 (Human breast adenocarcinoma epithelial cell) whole cell lysates

    Lysates/proteins at 15 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

    Predicted band size: 18 kDa


    Exposure time: 3 minutes


    This data was developed using ab109199, the same antibody clone in a different buffer formulation.

    Blocking and dilution buffer: 5% NFDM/TBST.

  • Western blot - Anti-p21 antibody [EPR3993] - BSA and Azide free (ab215971)
    Western blot - Anti-p21 antibody [EPR3993] - BSA and Azide free (ab215971)
    Lane 1 : Anti-p21 antibody [EPR3993] (ab109199) (1.4ug/ul)
    Lane 2 : Anti-p21 antibody [EPR18021] (ab188224) (1.4ug/ul)

    Lane 1 : RAW264.7 (Mouse Abelson murine leukemia virus-induced tumor macrophage) whole cell lysates
    Lane 2 : Neuro-2a (Mouse neuroblastoma neuroblast) whole cell lysates

    Lysates/proteins at 15 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

    Predicted band size: 18 kDa



    This data was developed using ab109199, the same antibody clone in a different buffer formulation.

  • Western blot - Anti-p21 antibody [EPR3993] - BSA and Azide free (ab215971)
    Western blot - Anti-p21 antibody [EPR3993] - BSA and Azide free (ab215971)

    This data was developed using ab109199, the same antibody clone in a different buffer formulation.

    Lane 1 Wild-type DLD-1 cell lysate (20 µg)

    Lane 2 Wild-type DLD-1 20 µM 2,3-DCPE for 16hrs treated cell lysate (20 µg)

    Lane 3 p21 knockout DLD-1 cell lysate (20 µg)

    Lane 4 p21 knockout 20 µM 2,3-DCPE for 16hrs DLD-1 cell lysate (20 µg)

    Lane 5: HT1080 cell lysate (20 µg)

    Lanes 1 - 5 Merged signal (red and green). Green - ab109199 observed at 20 kDa. Red - loading control, ab8245, observed at 37 kDa.

    This western blot image is a comparison between ab109119 and a competitor's top cited rabbit polyclonal antibody.

  • Western blot - Anti-p21 antibody [EPR3993] - BSA and Azide free (ab215971)
    Western blot - Anti-p21 antibody [EPR3993] - BSA and Azide free (ab215971)
    Anti-p21 antibody [EPR3993] (ab109199) at 1/1000 dilution (purified) + PC-12 cell lysate at 10 µg

    Secondary
    Peroxidase-conjugated goat anti-rabbit IgG (H+L) at 1/1000 dilution

    Predicted band size: 18 kDa
    Observed band size: 21 kDa why is the actual band size different from the predicted?



    This data was developed using ab109199, the same antibody clone in a different buffer formulation.

    Blocking and dilution buffer: 5% NFDM/TBST.

  • Anti-p21 antibody [EPR3993] - BSA and Azide free (ab215971)
    Anti-p21 antibody [EPR3993] - BSA and Azide free (ab215971)

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