Astana Biomed Group, an authorized Abcam distributor in Central Asia

Anti-PD1 (phospho Y248) antibody [EPR19821] - BSA and Azide free (ab234960)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR19821] to PD1 (phospho Y248) - BSA and Azide free
  • Suitable for: WB, Dot blot, IP
  • Reacts with: Human

Overview

  • Product name

    Anti-PD1 (phospho Y248) antibody [EPR19821] - BSA and Azide free
    See all PD1 primary antibodies

  • Description

    Rabbit monoclonal [EPR19821] to PD1 (phospho Y248) - BSA and Azide free

  • Host species

    Rabbit

  • Tested Applications & Species

    Application Species
    IP
    Human
    WB
    Human
    See all applications and species data

  • Immunogen

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

  • Positive control

    • WB: HEK-293T transfected with a PD1 (WT) plus a 25 kDa fusion protein expression vector, then treated with 100 µM pervanadate for 10 minutes whole cell lysate.

  • General notes

    Ab234960 is the carrier-free version of ab206378. 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

    ab234960 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

  • Immunoprecipitation - Anti-PD1 (phospho Y248) antibody [EPR19821] - BSA and Azide free (ab234960)
    Immunoprecipitation - Anti-PD1 (phospho Y248) antibody [EPR19821] - BSA and Azide free (ab234960)

    PD1 (phospho Y248) was immunoprecipitated from 0.35 mg of HEK-293T (human epithelial cell line from embryonic kidney transformed with large T antigen) transfected with a PD1 (WT) plus 25 kDa fusion protein expression vector, then treated with 100 μM pervanadate for 10 minutes whole cell lysate, with ab206378 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab206378 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366) was used for detection at 1/5000 dilution.

    Lane 1: HEK-293T transfected with a PD1 (WT) plus 25 kDa fusion protein expression vector, then treated with 100 μM pervanadate for 10 minutes whole cell lysate 10 μg (Input).

    Lane 2: ab206378 IP in HEK-293T transfected with a PD1 (WT) plus 25 kDa fusion protein expression vector, then treated with 100 μM pervanadate for 10 minutes whole cell lysate.

    Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab206378 in HEK-293T transfected with a PD1 (WT) plus 25 kDa fusion protein expression vector, then treated with 100 μM pervanadate for 10 minutes whole cell lysate.

    Exposure time: 30 seconds
    Blocking and diluting buffer and concentration: 5% NFDM/TBST

    The observed bands around 75 kDa comprise PD-1 plus the 25 kDa fusion protein. The plasmids were kindly provided by our collaborator Dr. Liang Chen, NIBS.

     

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab206378).

  • Dot Blot - Anti-PD1 (phospho Y248) antibody [EPR19821] - BSA and Azide free (ab234960)
    Dot Blot - Anti-PD1 (phospho Y248) antibody [EPR19821] - BSA and Azide free (ab234960)

    Dot blot analysis of PD1 (phospho Y248) labeled with ab206378 at 1/1000 dilution.

    Lane 1: PD1 (phospho Y248) peptide.
    Lane 2: PD1 non-phospho peptide.

    Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution was used as secondary antibody.

    Blocking/Dilution buffer: 5% NFDM/TBST.

    Exposure time: 1 minute.

    The blot was developed on a BIO-RAD® ChemiDoc™ MP instrument.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab206378).

  • Western blot - Anti-PD1 (phospho Y248) antibody [EPR19821] - BSA and Azide free (ab234960)
    Western blot - Anti-PD1 (phospho Y248) antibody [EPR19821] - BSA and Azide free (ab234960)
    All lanes : Anti-PD1 (phospho Y248) antibody [EPR19821] (ab206378) at 1/1000 dilution

    Lane 1 : Untreated-HEK-293T (human epithelial cell line from embryonic kidney transformed with large T antigen) whole cell lysate
    Lane 2 : HEK-293T transfected with a PD1 (WT) plus a 25 kDa fusion protein expression vector, then treated with 100 µM pervanadate for 10 minutes whole cell lysate
    Lane 3 : HEK-293T transfected with PD1 (WT) plus a 25 kDa fusion protein expression vector, then treated with 100 µM pervanadate for 10 minutes, whole cell lysate (20 µg). Following the transfer to PVDF, the membrane was treated with alkaline phosphatase for 1h at 37°C.

    Lysates/proteins at 20 µg per lane.

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

    Predicted band size: 32 kDa
    Additional bands at: 70-75 kDa (possible non-specific secondary antibody binding)


    Exposure time: 30 seconds


    Blocking/Dilution buffer: 5% NFDM/TBST.

    The observed bands at around 75 kDa are PD-1 plus the 25 kDa fusion protein. The expression profile observed is consistent with what has been described in the literature (PMID: 22641383). The plasmids were kindly provided by our collaborator Dr. Liang Chen NIBS.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab206378).

     

  • Anti-PD1 (phospho Y248) antibody [EPR19821] - BSA and Azide free (ab234960)
    Anti-PD1 (phospho Y248) antibody [EPR19821] - BSA and Azide free (ab234960)

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