Rabbit Recombinant Monoclonal PD1 phospho Y248 antibody. Carrier free. Suitable for IP, Dot, WB and reacts with Human, Synthetic peptide, Transfected cell lysate - Human samples.
IgG
Rabbit
pH: 7.2 - 7.4
Constituents: PBS
Liquid
Monoclonal
IP | Dot | WB | |
---|---|---|---|
Human | Tested | Expected | Tested |
Synthetic peptide | Not recommended | Tested | Not recommended |
Transfected cell lysate - Human | Not recommended | Not recommended | Tested |
Species | Dilution info | Notes |
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Species Human | Dilution info - | Notes - |
Species | Dilution info | Notes |
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Species Synthetic peptide, Transfected cell lysate - Human | Dilution info - | Notes - |
Species | Dilution info | Notes |
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Species Synthetic peptide | Dilution info - | Notes - |
Species | Dilution info | Notes |
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Species Human | Dilution info Use at an assay dependent concentration. | Notes - |
Species | Dilution info | Notes |
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Species Transfected cell lysate - Human | Dilution info - | Notes - |
Species | Dilution info | Notes |
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Species Transfected cell lysate - Human, Human | Dilution info - | Notes - |
Species | Dilution info | Notes |
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Species Synthetic peptide | Dilution info - | Notes - |
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Inhibitory receptor on antigen activated T-cells that plays a critical role in induction and maintenance of immune tolerance to self (PubMed:21276005). Delivers inhibitory signals upon binding to ligands CD274/PDCD1L1 and CD273/PDCD1LG2 (PubMed:21276005). Following T-cell receptor (TCR) engagement, PDCD1 associates with CD3-TCR in the immunological synapse and directly inhibits T-cell activation (By similarity). Suppresses T-cell activation through the recruitment of PTPN11/SHP-2: following ligand-binding, PDCD1 is phosphorylated within the ITSM motif, leading to the recruitment of the protein tyrosine phosphatase PTPN11/SHP-2 that mediates dephosphorylation of key TCR proximal signaling molecules, such as ZAP70, PRKCQ/PKCtheta and CD247/CD3zeta (By similarity).The PDCD1-mediated inhibitory pathway is exploited by tumors to attenuate anti-tumor immunity and escape destruction by the immune system, thereby facilitating tumor survival (PubMed:28951311). The interaction with CD274/PDCD1L1 inhibits cytotoxic T lymphocytes (CTLs) effector function (PubMed:28951311). The blockage of the PDCD1-mediated pathway results in the reversal of the exhausted T-cell phenotype and the normalization of the anti-tumor response, providing a rationale for cancer immunotherapy (PubMed:22658127, PubMed:25034862, PubMed:25399552).
Programmed cell death protein 1, Protein PD-1, hPD-1, PDCD1, PD1
Rabbit Recombinant Monoclonal PD1 phospho Y248 antibody. Carrier free. Suitable for IP, Dot, WB and reacts with Human, Synthetic peptide, Transfected cell lysate - Human samples.
IgG
Rabbit
pH: 7.2 - 7.4
Constituents: PBS
Liquid
Monoclonal
Yes
EPR19821
Affinity purification Protein A
Blue Ice
+4°C
Do Not Freeze
ab234960 is the carrier-free version of Anti-PD1 (phospho Y248) antibody [EPR19821] ab206378.
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 monoclonal antibody, which offers several advantages including:
For more information, read more on recombinant antibodies.
Our carrier-free antibodies are typically supplied in a PBS-only formulation, purified and free of BSA, sodium azide and glycerol. The carrier-free buffer and high concentration allow for increased conjugation efficiency.
This conjugation-ready format is designed for use with fluorochromes, metal isotopes, oligonucleotides, and enzymes, which makes them ideal for antibody labelling, functional and cell-based assays, flow-based assays (e.g. mass cytometry) and Multiplex Imaging applications.
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.
This product is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm, without the need for antibody preparation. Maxpar® is a trademark of Fluidigm Canada Inc.
This supplementary information is collated from multiple sources and compiled automatically.
PD1 also known as Programmed Cell Death Protein 1 or PDCD1 is a transmembrane protein that plays a critical role in regulating immune responses. It has a mass of approximately 55 kDa. PD1 is expressed on the surface of T cells B cells and some myeloid cells. PD1’s expression increases upon activation of these immune cells assisting in maintaining peripheral tolerance. Researchers often use PD1 mouse models and chimeric antibodies to explore the function of PD1 for experimental purposes. Antibodies such as anti-PD1 such as EH12.2H7 help in blocking PD1 interaction to study its role further.
PD1 serves as an inhibitory receptor acting as a checkpoint in the immune system. It becomes part of an immune-suppressive complex when it binds with its ligands PD-L1 or PD-L2 which are expressed on various cell types including some tumor cells. This interaction suppresses the proliferation of T cells and cytokine production contributing to immune homeostasis. By controlling T cell activity PD1 limits autoimmunity but can also reduce the immune system's capability to attack cancer cells.
PD1 functions in the immune checkpoint pathway a critical regulatory circuit in immune regulation. The engagement of PD1 with its ligands initiates a cascade that inhibits the function and proliferation of T cells through downstream SHP-2 phosphatase activity. This pathway frequently involves other regulatory proteins like CTLA-4 and is an important mechanism by which the body modulates immune responses. Related pathways often intersect with those involving T cell receptor signaling and contribute to the overall modulation of immune activity.
PD1 has a significant role in cancer and autoimmune disorders. PD1 expression can allow tumors to evade immune surveillance making PD1 a target for cancer therapies such as anti-PD1 antibodies which aim to block PD1 and restore T cell activity. The interaction of PD1 with cancer-related proteins like PD-L1 facilitates tumor immune evasion. In autoimmune disorders PD1’s regulation of immune balance can become dysregulated leading to persistent immune activation and tissue damage. Understanding PD1 and its interaction with proteins such as PD-L1 helps in developing therapeutic strategies for both cancer and autoimmune conditions.
We have tested this species and application combination and it works. It is covered by our product promise.
We have not tested this specific species and application combination in-house, but expect it will work. It is covered by our product promise.
This species and application combination has not been tested, but we predict it will work based on strong homology. However, this combination is not covered by our product promise.
We do not recommend this combination. It is not covered by our product promise.
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In the unlikely event of one of our products not working as expected, you are covered by our product promise.
Full details and terms and conditions can be found here:
Terms & Conditions.
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 Anti-PD1 (phospho Y248) antibody [EPR19821] ab206378 at 1/30 dilution. Western blot was performed from the immunoprecipitate using Anti-PD1 (phospho Y248) antibody [EPR19821] ab206378 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (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: Anti-PD1 (phospho Y248) antibody [EPR19821] 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 (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) instead of Anti-PD1 (phospho Y248) antibody [EPR19821] 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 (Anti-PD1 (phospho Y248) antibody [EPR19821] ab206378).
All lanes: Immunoprecipitation - Anti-PD1 (phospho Y248) antibody [EPR19821] (Anti-PD1 (phospho Y248) antibody [EPR19821] ab206378)
Predicted band size: 32 kDa
Observed band size: 70-75 kDa
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 (Anti-PD1 (phospho Y248) antibody [EPR19821] ab206378).
All lanes: Western blot - Anti-PD1 (phospho Y248) antibody [EPR19821] - BSA and Azide free (ab234960) at 1/1000 dilution
Lane 1: Untreated-HEK-293T (human epithelial cell line from embryonic kidney transformed with large T antigen) whole cell lysate at 20 µg
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 at 20 µg
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. at 20 µg
All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/20000 dilution
Predicted band size: 32 kDa
Observed band size: 70-75 kDa
Exposure time: 30s
Dot blot analysis of PD1 (phospho Y248) labeled with Anti-PD1 (phospho Y248) antibody [EPR19821] ab206378 at 1/1000 dilution.
Lane 1: PD1 (phospho Y248) peptide.
Lane 2: PD1 non-phospho peptide.
Goat Anti-Rabbit IgG H&L (HRP) (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 (Anti-PD1 (phospho Y248) antibody [EPR19821] ab206378).
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