Anti-PD1 (phospho Y248) antibody [EPR19821] (ab206378)
![Anti-PD1 (phospho Y248) antibody [EPR19821] (ab206378)](https://www.abcam.com/ps/products/206/ab206378/Images/ab206378-309856-anti-pd1-phospho-y248-antibody-epr19821-immunoprecipitation.jpg "Anti-PD1 (phospho Y248) antibody [EPR19821] (ab206378)")
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)
- Suitable for: WB, Dot blot, IP
- Reacts with: Human
Overview
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Product name
Anti-PD1 (phospho Y248) antibody [EPR19821]
See all PD1 primary antibodies -
Description
Rabbit monoclonal [EPR19821] to PD1 (phospho Y248) -
Host species
Rabbit -
Tested Applications & Species
See all applications and species dataApplication Species ICC/IF Recombinant fragmentIP HumanWB Human
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Immunogen
Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
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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. IP: HEK-293T transfected with PD1 (WT) plus 25 kDa fusion protein expression vector, then treated with 100 µM pervanadate for 10 minutes whole cell lysate.
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General notes
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
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing the problem with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation.
One factor contributing to the crisis is the use of antibodies that are not suitable. This can lead to misleading results and the use of incorrect data informing project assumptions and direction. To help address this challenge, we have introduced an application and species grid on our primary antibody datasheets to make it easy to simplify identification of the right antibody for your needs.
Learn more here.
Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle. -
Storage buffer
pH: 7.2
Preservative: 0.01% Sodium azide
Constituents: PBS, 40% Glycerol, 0.05% BSA -
Concentration information loading... -
Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR19821 -
Isotype
IgG -
Research areas
Images
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Immunoprecipitation - Anti-PD1 (phospho Y248) antibody [EPR19821] (ab206378)
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/TBSTThe 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.
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![Western blot - Anti-PD1 (phospho Y248) antibody [EPR19821] (ab206378)](https://www.abcam.com/ps/products/206/ab206378/Images/ab206378-309855-anti-pd1-phospho-y248-antibody-epr19821-western-blot.jpg)
Western blot - Anti-PD1 (phospho Y248) antibody [EPR19821] (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 transfected with a PD1 (WT) plus a 25 kDa fusion protein expression vector
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
Observed band size: 70-75 kDa why is the actual band size different from the predicted?
Exposure time: 30 secondsBlocking/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.
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![Dot Blot - Anti-PD1 (phospho Y248) antibody [EPR19821] (ab206378)](https://www.abcam.com/ps/products/206/ab206378/Images/ab206378-309854-anti-pd1-phospho-y248-antibody-epr19821-dot-blot.jpg)
Dot Blot - Anti-PD1 (phospho Y248) antibody [EPR19821] (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.
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![Anti-PD1 (phospho Y248) antibody [EPR19821] (ab206378)](https://www.abcam.com/ps/products/206/ab206378/Images/ab206378-5-benefits-of-recombinant-antibodies.png)
Anti-PD1 (phospho Y248) antibody [EPR19821] (ab206378)
