Anti-PD1/PD-1 antibody, human IgG1, recombinant

Product ID: anti-PD-1 hIgG1 (SKU#: MAB1732)

Price:

$449.00

Description

Programmed death-1 (PD-1), also known as programmed cell death 1 (PDCD1) or CD279, is a single pass type I transmembrane glycoprotein of the Ig superfamily. It is an immunoreceptor in the CD28/CTLA-4 family with an IgV-type extracellular domain showing 21–33% sequence identity with CTLA-4, CD28 and ICOS. Members of the CD28/CTLA-4 family have been shown to either promote T cell activation (CD28 and ICOS) or inhibit T cell activation (CTLA4 and PD1). The cytoplasmic domain of PD-1 contains two tyrosine residues, a membrane-proximal immunoreceptor tyrosine-based inhibitory motif (ITIM) and an immunoreceptor tyrosine-based switch motif (ITSM). ITIM is widely found in immunoinhibitory receptors and the membrane-proximal tyrosine residue may play a central role for the inhibitory function of PD-1. PD-1 negatively regulates antigen receptor signaling by recruiting protein tyrosine phosphatase upon engaging with either of two ligands, PD-L1 or PD-L2. It inhibits the T-cell proliferation and production of related cytokines such as IL-1, IL-4, IL-10 and IFN-γ. PD-1 is involved in lymphocyte clonal selection and peripheral tolerance, and thus contributes to the prevention of autoimmune diseases. As a negative regulator of T- cell effector mechanisms, PD-1 may lead to suppression of anti-tumor immunity. Blockade of PD-1 inhibitory activity with antibodies enhances anti-tumor immunity in vivo. PD-1 has been proposed as a promising target for cancer immunotherapy.

Gene Symbol: PD-1; CD279, PDCD1, PD1, SLEB2, hPD-1, hPD-l, hSLE1

NCBI Gene ID: 5133

Uniprot Entry: Q15116

Construct Details: The recombinant antibody is expressed as the combination of a heavy chain (HC) containing VH from anti-PD-1 mAb and CH1-3 region of human IgG1 and a light chain (LC) encoding VL from anti-PD-1 mAb and CL of human kappa light chain. Exists as a disulfide linked dimer of the HC and LC hetero-dimer under non-reducing condition.

Source: Mammalian cells stably expressing anti-PD-1 HC and LC and growing in chemical-defined media with no animal component or antibiotics.

M.W.: Calculated molecular mass (kDa): 146 (49 kDa for a single HC, 24 kDa for a single LC); Estimated by SDS-PAGE under non-reducing condition(kDa): 150.

Purity: >95% judged by SDS-PAGE under reducing condition (see the gel image above)

Formulation: Supplied at 1.0 mg/ml in sterile PBS pH7.4 without any carrier protein or preservative (the concentration was determined by UV absorption with an A280nm of 1.0 for 0.73 mg/ml of IgG and verified by SDS-PAGE and Coomassie blue staining).

Endotoxin Level: <0.1 EU per 1 μg of purified recombinant protein determined by the LAL method

Biological Activity:

Binds to PD-1 extracellular domain (SKU#: FCL0761 and FCL0763) with high affinity (K_D< 1 nM as determined by ELISA - see the attached technical data). Recognizes cell surface PD-1 by flow cytometry, IHC staining and fluorescent microscopy. Blocks PD-1’s binding to its ligands, PD-L1 and PD-L2, and neutralizes their signaling activities. Enhances anti-tumor immunity in vivo. Binds PD-1 with the same specificity as anti-PD-1 monoclonal antibodies, mouse IgG2a (SKU#MAB0761) and rabbit IgG (SKU#MAB0763).

FOR RESEARCH USE ONLY. NOT FOR DIAGNOSTIC OR THERAPEUTIC USE IN HUMAN.

Restriction: This product is not transferable or re-sellable. Customer obtain no right to transfer, assign, or sublicense its use rights, or to transfer, resell, package, or otherwise distribute the product, or to use the product for the benefit of any third party or for any commercial purpose. Customer may only use the product in compliance with applicable local, state and federal laws, regulations and rules. Customer may not directly or indirectly use the product or allow the transfer, transmission, export or re-export of all or any part of the product in violation of any export control law or regulation of the united states or any other relevant jurisdiction. Your use of this product constitutes acceptance of the terms of this limited use agreement. Please refer to our “terms & conditions” for details. If you are not willing to accept the limitation of this agreement, G&P Biosciences will accept return of the product for a full/partial refund.

Recombinant PD-1 ECD protein (SKU#FCL0761) was immobilized at 100 ng/well on a 96-well ELISA plate and incubated with serially diluted anti-PD-1 human IgG1 monoclonal antibody (SKU#MAB1732). Anti-CTLA4 (SKU#MAB1718) human IgG1 monoclonal antibody was used as a control to demonstrate the specificity of PD-1 binding under the experimental conditions. The bound antibodies were detected by goat anti-human IgG HRP conjugated secondary antibody. The ELISA binding curve was fit and K_D was calculated for anti-PD-1:PD-1 binding affinity/avidity to be 23 pM.

PD-1 ELISA

FOR RESEARCH USE ONLY. NOT FOR DIAGNOSTIC OR THERAPEUTIC USE IN HUMAN.

The product is shipped at 4°C. Upon receipt, centrifuge the product briefly before opening the vial. It is recommended to store small aliquots at the temperature below –20°C for long-term storage and the product is stable for 3 months. The undiluted protein can be stored at 4°C for no more than 2 weeks.

Use a manual defrost freezer and avoid repeated freeze-thaw cycles.

FOR RESEARCH USE ONLY. NOT FOR DIAGNOSTIC OR THERAPEUTIC USE IN HUMAN.

1. Nat. Med. 5:1365-1369, 1999

2. J. Exp. Med. 192:1027-1034, 2000

3. Nature Med. 8: 793-800, 2002

4. Nature Med. 9: 562-567, 2003

Product datasheet (pdf) can be downloaded here: MAB1732-PDS.pdf