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Key Product Details

Validated by

Knockout/Knockdown, Independent Antibodies, Biological Validation

Species Reactivity

Human, Mouse, Rat

Applications

Dual RNAscope ISH-IHC, ELISA, Flow Cytometry, Immunocytochemistry/ Immunofluorescence, Immunofluorescence, Immunohistochemistry, Immunohistochemistry Whole-Mount, Immunohistochemistry-Frozen, Immunohistochemistry-Paraffin, Immunoprecipitation, Knockdown Validated, Western Blot

Label

Unconjugated

Antibody Source

Polyclonal Rabbit IgG

Format

BSA Free

Concentration

1 mg/ml

Product Summary for PD-L1 Antibody - BSA Free

Immunogen

Antibody was raised against a 17 amino acid synthetic peptide from near the center of human PD-L1. The immunogen is located within amino acids 60-110 of PD-L1.

Specificity

PD-L1 antibody has no cross-reactivity to PD-L2.

Clonality

Polyclonal

Host

Rabbit

Isotype

IgG

Theoretical MW

37 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.

Scientific Data Images for PD-L1 Antibody - BSA Free

Knockdown Validated: PD-L1 Antibody [NBP1-76769] - Validation with PD-L1 siRNA Knockdown. HeLa cells were transfected with control siRNAs (lane 1) or PD-L1 siRNAs (lane 2). Loading: 10 ug of HeLa whole cell lysates per lane. Antibodies: NBP1-76769 (2 ug/mL) and GAPDH (0.02 ug/mL), 1 h incubation at RT in 5% NFDM/TBST. Secondary: Goat anti-rabbit IgG HRP conjugate at 1:10000 dilution.
Western Blot: PD-L1 Antibody [NBP1-76769] - Independent Antibody Validation (IAV) via Protein Expression ProfileLoading: 15 ug of lysates per lane.Antibodies: NBP1-76769 (2 ug/mL), PD-L1 (2 ug/mL), and beta-actin (1 ug/mL), 1 h incubation at RT in 5% NFDM/TBST.Secondary: Goat anti-rabbit and or anti-mouse IgG HRP conjugate at 1:10000 and 1:5000 dilution, respectively.
Immunohistochemistry-Paraffin: PD-L1 Antibody [NBP1-76769] - CD274 (PD-L1) expression in the HNSCC patients from the TCGA database , HNSCC tissue samples and the HNSCC cells. Immunostaining of PD-L1 obtained from HNSCC tumor cells, immune cells and tumor margin tissues in HNSCC tissue samples (magnification A-200, scale bars 50AI1/4m) : low tumor staining; moderate tumor staining; high tumor staining have been observed. Image collected and cropped by Citeab from the following publication (Lactoferricin B reverses cisplatin resistance in head and neck squamous cell carcinoma cells through targeting PD-L1. Cancer Med (2018)) licensed under a CC-BY license.

Applications for PD-L1 Antibody - BSA Free

Application
Recommended Usage

ELISA

1:100 - 1:2000

Flow Cytometry

0.5 ug/ml

Immunocytochemistry/ Immunofluorescence

1-5 ug/ml

Immunofluorescence

20 ug/ml

Immunohistochemistry

1:10-1:500

Immunohistochemistry-Paraffin

1:10-1:500

Western Blot

1:1000
Application Notes
Use in Immunohistochemistry Whole-Mount reported in scientific literature (PMID:34944780).Use in ICC/IF reported in scientific literature (PMID:33220359) Use in IHC-Frozen was reported in scientific literature (PMID: 28402953). Use in immunoprecipitation reported in scientific literature (PMID: 28978117)..
Please Note: Optimal dilutions of this antibody should be experimentally determined.

Reviewed Applications

Read 5 reviews rated 3.2 using NBP1-76769 in the following applications:

Published Applications

Read 40 publications using NBP1-76769 in the following applications:

Formulation, Preparation, and Storage

Purification

Peptide affinity purified

Formulation

PBS

Format

BSA Free

Preservative

0.02% 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

Programmed-death ligand 1 (PD-L1), also known as CD274 and B7-H1, is a 33 kDa type I glycoprotein containing 290 amino acids (aa) belonging to the protein B7 family and serves as part of an immune checkpoint (1,2). PD-L1 contains an Ig-V and Ig-C-like extracellular domain, a transmembrane domain, and a cytoplasmic tail lacking canonical signaling motifs (2,3). PD-L1 is the ligand that binds the receptor programmed-death 1 (PD-1) which is highly expressed on active T cells (1-3). PD-L1 is typically upregulated by tumor cells and antigen presenting cells (APCs), but also expressed on T cells, B cells, macrophages, dendritic cells (DC), mast cells, and some non-immune cell types (1-3). In addition to the membrane-bound, PD-L1 is released from cells both in soluble form and bound to extracellular vesicles (4).

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

UniProt

Product Documents for PD-L1 Antibody - BSA Free

Certificate of Analysis

To download a Certificate of Analysis, please enter a lot number in the search box below.

Product Specific Notices for PD-L1 Antibody - BSA Free

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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FAQs for PD-L1 Antibody - BSA Free

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  • Q: What can I use as positive controls for B7-H1 (NBP1-76769) other than the myocardial tissue?

    A: For CD274 (B7-H1), this protein is highly expressed in the heart, skeletal muscle, placenta and lung ( uniprot site). It can also be found at lower levels in other tissues as well: protein atlas site.

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