Recombinant
RabMAb

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

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
    Suitable for: WB, Dot blot, IPmore details
  • Species reactivity
    Reacts with: Human
  • Immunogen

    Synthetic peptide within Human PD1 aa 200 to the C-terminus (phospho Y248). The exact sequence is proprietary.
    Database link: Q15116

  • 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

    The formulation and the concentration of this product is compatible for metal-conjugation for mass cytometry (CyTOF®).

    ab234960 is a PBS-only buffer format of ab206378. Please refer to ab206378 for recommended dilutions, protocols, and image data.

    Use our conjugation kits for antibody conjugates that are ready-to-use in as little as 20 minutes with <1 minute hands-on-time and 100% antibody recovery: available for fluorescent dyes, HRP, biotin and gold.

    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 ab234960 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
WB Use at an assay dependent concentration. Predicted molecular weight: 32 kDa.
Dot blot Use at an assay dependent concentration.
IP Use at an assay dependent concentration.

Target

  • Function
    Possible cell death inducer, in association with other factors.
  • Involvement in disease
    Genetic variation in PDCD1 is associated with susceptibility to systemic lupus erythematosus type 2 (SLEB2) [MIM:605218]. Systemic lupus erythematosus is a chronic, inflammatory and often febrile multisystemic disorder of connective tissue. It affects principally the skin, joints, kidneys and serosal membranes. It is thought to represent a failure of the regulatory mechanisms of the autoimmune system.
  • Sequence similarities
    Contains 1 Ig-like V-type (immunoglobulin-like) domain.
  • Developmental stage
    Induced at programmed cell death.
  • Cellular localization
    Membrane.
  • Information by UniProt
  • Database links
  • Alternative names
    • CD279 antibody
    • CD279 antigen antibody
    • hPD 1 antibody
    • hPD l antibody
    • hPD-1 antibody
    • hSLE1 antibody
    • PD 1 antibody
    • PD-1 antibody
    • PD1 antibody
    • PDCD 1 antibody
    • PDCD1 antibody
    • PDCD1_HUMAN antibody
    • Programmed cell death 1 antibody
    • Programmed cell death 1 protein antibody
    • Programmed cell death protein 1 antibody
    • Protein PD 1 antibody
    • Protein PD-1 antibody
    • SLEB2 antibody
    • Systemic lupus erythematosus susceptibility 2 antibody
    see all

Images

  • 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 secondary antibody (HRP) (ab131366) was used as the secondary antibody 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 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).

  • 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).

     

References

ab234960 has not yet been referenced specifically in any publications.

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Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

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