Mouse PD-L1 protein, ECD (extracellular domain), Fc-fusion, recombinant

Product ID: mPD-L1-Fc (SKU#: FCL1846)

Price:

$598.00

Description

PD-L1 (programmed cell death 1 ligand 1) or CD274 is a 290-amino acid single pass type I transmembrane protein of the Ig (immunoglobulin) superfamily. It contains one Ig V-like and one Ig C-like domains in the extracellular region, and a 30-amino acid cytoplasmic tail. It was originally cloned as a homolog of B7 (termed B7-H1). It is 20% and 15% identical to B7-1 (CD80) and B7-2 (CD86), respectively. It was also cloned as a ligand (termed PDCD1L1 or PD-L1) that binds to PD-1 (CD279) and B7-1 (CD80), but not to other B7 family members, such as CTLA4 (CD152), CD28, or ICOS (CD278). PD-L1 or CD274 is involved in the T cell costimulatory signal, essential for T-cell proliferation and production of IL10 and IFNγ. Interaction with PD-1, which has a cytoplasmic immunoreceptor tyrosine-based inhibitory motif (ITIM), inhibits T-cell proliferation and cytokine production. PD-L1 is up-regulated on T- and B-cells, dendritic cells, keratinocytes and monocytes after LPS and IFNγ activation or by surface Ig cross-linking. It is expressed in many types of freshly isolated human cancers but not in most normal tissues. Blockade of PD-L1 upregulates IL12 expression and enhances antitumor immunity in mice bearing ovarian tumors. Therefore, PD-L1 has been proposed as a target for cancer immunotherapy through pre-activated T cells that could be enhanced by blockade of PD-L1-induced inhibitory signaling.

Gene Symbol: PD-L1; CD274; B7-H1; B7H1; PDL1; PDCD1L1; PDCD1LG1

NCBI Gene ID: 60533

Uniprot Entry: Q9EP73

Construct Details: The recombinant mouse PD-L1/CD274 ECD is expressed as a 458 amino acid protein consisting of Phe19 - Thr238 region of mouse PD-L1/CD274 (UniProt accession #Q9EP73) and a C-terminal Fc from human IgG1, which exists as a dimer under non-reducing conditions.

Source: Mammalian cells stably expressing mouse PD-L1-Fc and growing in chemical-defined media with no animal components or antibiotics

Deduced Amino Acid Sequence:

10 20 30 40 50 60

FTITAPKDLY VVEYGSNVTM ECRFPVEREL DLLALVVYWE KEDEQVIQFV AGEEDLKPQH

70 80 90 100 110 120

SNFRGRASLP KDQLLKGNAA LQITDVKLQD AGVYCCIISY GGADYKRITL KVNAPYRKIN

130 140 150 160 170 180

QRISVDPATS EHELICQAEG YPEAEVIWTN SDHQPVSGKR SVTTSRTEGM LLNVTSSLRV

190 200 210 220 230 240

NATANDVFYC TFWRSQPGQN HTAELIIPEL PATHPPQNRT STTENLYFQG STGTHTCPPC

250 260 270 280 290 300

PAPELLGGPS VFLFPPKPKD TLMISRTPEV TCVVVDVSHE DPEVKFNWYV DGVEVHNAKT

310 320 330 340 350 360

KPREEQYNST YRVVSVLTVL HQDWLNGKEY KCKVSNKALP APIEKTISKA KGQPREPQVY

370 380 390 400 410 420

TLPPSREEMT KNQVSLTCLV KGFYPSDIAV EWESNGQPEN NYKTTPPVLD SDGSFFLYSK

430 440 450

LTVDKSRWQQ GNVFSCSVMH EALHNHYTQK SLSLSPGK

M.W.:

Calculated molecular mass 51.5 kDa; estimated by SDS-PAGE under reducing condition 60-65 kDa suggesting that the protein is highly glycosylated

Calculated PI: 6.00

Calculated Extinction Coefficients (M-1 cm-1, at 280nm): 67435

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

Formulation: Supplied at 0.5 mg/ml in sterile PBS pH7.4 (carrier and preservative free).

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

Biological Activity: Binds to its receptor PD-1 and anti-PD-L1 monoclonal antibody (SKU#: MAB0199, MAB0784, MAB0781 - see Technical Data). Blocks PD-L1’s binding to its receptor and mediated signaling activity. Shows cross-species binding to human PD-1 (SKU#FCL0763) and human CD80 (SKU#FCL0723) in a functional ELISA (see Technical Data).

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.

Technical Data 1: Anti-m,hPD-L1 Monoclonal Antibody Binding Affinity Determination by ELISA

Recombinant human PD-L1-Fc (hPD-L1, SKU#FCL0781), mouse PD-L1-Fc (mPD-L1, SKU#FCL1846), human PD-L2-Fc (hPD-L2, SKU#FCL0783), and mouse PD-L2-Fc (mPD-L2, SKU#FCL1818) proteins was immobilized at 100 ng/well on a 96-well NuncMaxisorp plate for ELISA with serially diluted, biotinylated anti-PD-L1 monoclonal antibody, human IgG1(SKU#MAB0199B). PD-L2 proteins were used as a control to demonstrate the specificity of PD-L1 binding under the experimental conditions. As shown in the figure below, anti-PD-L1 mAb binds specifically to human and mouse PD-L1-Fc (but not to PD-L2-Fc) equally well with an apparent K_D for human PD-L1 of 38.1 ± 2.4 pM and an apparent K_D for mouse PD-L1 of 38.6± 2.5 pM.

Technical Data 2: Cross-species Interactions with Human PD-1 and CD80 by ELISA

Recombinant mouse PD-L1-Fc (SKU#FCL1846) was immobilized at 100 ng/well on a 96-well NuncMaxisorp plate for a functional ELISA with serially diluted, biotinylated human PD-1-Fc (PD-1-Fc-Biot, SKU#FCL0763B), biotinylated human CD80-Fc (CD80-Fc-Biot, SKU#FCL0723B), and biotinylated Fc alone (control Fc-Biot, SKU#FCL00029B). Fc protein alone was used as a negative control to demonstrate the specificity of PD-1:mPD-L1 and CD80:mPD-L1 interactions under the experimental conditions. Mouse PD-L1 shows cross-species reactivity to human PD-1 and CD80. The binding curve was fit and apparent K_D was calculated for PD-1: mPD-L1 binding to be 557 ± 63 ng/ml with a linear binding range between 150 and 2000 ng/ml, and apparent K_D for CD80:mPD-L1 binding to be 9486 ± 5500 ng/ml with a linear binding range between 1500 and 75000 ng/ml.

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: FCL1846-PDS.pdf