PD-L1 Antibody (PSH04-80)
Novus Biologicals, part of Bio-Techne | Catalog # NBP3-32731
Recombinant Monoclonal Antibody
Key Product Details
Species Reactivity
Mouse
Applications
Immunohistochemistry, Immunohistochemistry-Paraffin, Western Blot
Label
Unconjugated
Antibody Source
Recombinant Monoclonal Rabbit IgG Clone # PSH04-80
Concentration
1 mg/ml
Product Specifications
Immunogen
Recombinant protein within mouse PD-L1 aa 1-250 / 290. (Uniprot: Q9EP73)
Localization
Cell membrane, Early endosome membrane, Recycling endosome membrane.
Clonality
Monoclonal
Host
Rabbit
Isotype
IgG
Theoretical MW
33 kDa.
Disclaimer note: The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors.
Disclaimer note: The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors.
Scientific Data Images for PD-L1 Antibody (PSH04-80)
Western Blot: PD-L1 Antibody (PSH04-80) [NBP3-32731]
Western Blot: PD-L1 Antibody (PSH04-80) [NBP3-32731] - Western blot analysis of PD-L1 on different lysates with Rabbit anti-PD-L1 antibody (NBP3-32731) at 1/2,000 dilution.Lane 1: RAW264.7 cell lysate
Lane 2: RAW264.7 treated with 10ug/mL LPS for 8 hours cell lysate
Lane 3: J774A.1 cell lysate
Lane 4: J774A.1 treated with 1ug/mL LPS for 24 hours cell lysate
Lane 5: Mouse spleen tissue lysate
Lysates/proteins at 30 ug/Lane.
Predicted band size: 33 kDa
Observed band size: 45-60 kDa
Exposure time: 46 seconds; ECL;
4-20% SDS-PAGE gel.
Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (NBP3-32731) at 1/2,000 dilution was used in 5% NFDM/TBST at 4 overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody at 1/50,000 dilution was used for 1 hour at room temperature.
Immunohistochemistry: PD-L1 Antibody (PSH04-80) [NBP3-32731]
Immunohistochemistry: PD-L1 Antibody (PSH04-80) [NBP3-32731] - Immunohistochemical analysis of paraffin-embedded mouse thymus tissue with Rabbit anti-PD-L1 antibody (NBP3-32731) at 1/200 dilution.The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (NBP3-32731) at 1/200 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
Applications for PD-L1 Antibody (PSH04-80)
Application
Recommended Usage
Immunohistochemistry-Paraffin
1:200
Western Blot
1:2000
Formulation, Preparation, and Storage
Purification
Protein A purified
Formulation
PBS (pH7.4), 0.1% BSA and 40% Glycerol
Preservative
0.05% Sodium Azide
Concentration
1 mg/ml
Shipping
The product is shipped with polar packs. Upon receipt, store it immediately at the temperature recommended below.
Stability & Storage
Store at 4C short term. Aliquot and store at -20C long term. Avoid freeze-thaw cycles.
Background: PD-L1/B7-H1
PD-L1 binding with receptor PD-1 results in phosphorylation of in the inhibitory tyrosine-based switch motif (ITSM) domain of PD-1, which leads to recruitment of Src homology 2 domain-containing protein tyrosine-phosphatase 2 (SHP-2) and eventual downstream phosphorylation of spleen tyrosine kinase (Syk) and phospholipid inositol-3-kinase (PI3K) (1,3). Under normal conditions, the PD-L1/PD-1 signaling axis helps maintain immune tolerance and prevent destructive immune responses by inhibiting T cell activity such as proliferation, survival, cytokine production, and cytotoxic T lymphocyte (CTL) cytotoxicity (1-3). In the tumor microenvironment (TME), however, the PD-L1/PD-1 signaling axis is hijacked to promote tumor cell survival and limit anti-tumor immune response (1,3). More precisely, tumor cells can escape killing and immune surveillance due to T cell exhaustion and apoptosis (1-3).
Given the role the PD-L1/PD-1 signaling axis plays in tumor cells' ability to evade immune surveillance, it has become a target of several immunotherapeutic agents in recent years (3,5). Antibody immunotherapies that target these inhibitory checkpoint molecules has shown great promise for cancer treatment (3,5). PD-L1 and PD-1 blocking agents have been approved for treatment in a number of cancers including melanoma, non-small cell lung cancer (NSCLC), urothelial carcinoma, and Merkel-cell carcinoma (3,5). In many cancers the expression of PD-L1 in the TME has predictive value for response to blocking agents (3). Pembrolizumab, for example, is a PD-1 inhibitor that has been approved by the FDA as a second-line therapy for treatment of metastatic NSCLC in patients whose tumors express PD-L1 with a Tumor Proportion Score (TPS) greater than 1%, but also for first-line treatment in cases where patients' tumors expression PD-L1 with a TPS greater than 50%) (5). The most promising cancer immunotherapy treatments seem to point to combination therapy with both anti-cancer drugs (e.g. Gefitibin, Metformin, Etoposide) with PD-L1/PD-1 antibody blockade inhibitors (e.g. Atezolizumab, Nivolumab) (6).
References
1. Han, Y., Liu, D., & Li, L. (2020). PD-1/PD-L1 pathway: current researches in cancer. American journal of cancer research, 10(3), 727-742.
2. Jiang, Y., Chen, M., Nie, H., & Yuan, Y. (2019). PD-1 and PD-L1 in cancer immunotherapy: clinical implications and future considerations. Human vaccines & immunotherapeutics, 15(5), 1111-1122. https://doi.org/10.1080/21645515.2019.1571892
3. Sun, C., Mezzadra, R., & Schumacher, T. N. (2018). Regulation and Function of the PD-L1 Checkpoint. Immunity, 48(3), 434-452. https://doi.org/10.1016/j.immuni.2018.03.014
4. Cha, J. H., Chan, L. C., Li, C. W., Hsu, J. L., & Hung, M. C. (2019). Mechanisms Controlling PD-L1 Expression in Cancer. Molecular cell, 76(3), 359-370. https://doi.org/10.1016/j.molcel.2019.09.030
5. Tsoukalas, N., Kiakou, M., Tsapakidis, K., Tolia, M., Aravantinou-Fatorou, E., Baxevanos, P., Kyrgias, G., & Theocharis, S. (2019). PD-1 and PD-L1 as immunotherapy targets and biomarkers in non-small cell lung cancer. Journal of B.U.ON. : official journal of the Balkan Union of Oncology, 24(3), 883-888.
6. Gou, Q., Dong, C., Xu, H., Khan, B., Jin, J., Liu, Q., Shi, J., & Hou, Y. (2020). PD-L1 degradation pathway and immunotherapy for cancer. Cell death & disease, 11(11), 955. https://doi.org/10.1038/s41419-020-03140-2
Long Name
Programmed Death Ligand 1
Alternate Names
B7-H1, B7H1, CD274, PDCD1L1, PDCD1LG1, PDL1
Gene Symbol
CD274
Additional PD-L1/B7-H1 Products
Product Documents for PD-L1 Antibody (PSH04-80)
Product Specific Notices for PD-L1 Antibody (PSH04-80)
This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.
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