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Key Product Details

Species Reactivity

Human, Mouse

Applications

Flow Cytometry, Immunocytochemistry/ Immunofluorescence, Immunohistochemistry, Immunohistochemistry-Frozen, In vivo assay, Western Blot

Label

Unconjugated

Antibody Source

Monoclonal Rat IgG2a Lambda Clone # MIH5

Format

BSA Free

Concentration

0.5 mg/ml

Product Summary for PD-L1 Antibody (MIH5) - BSA Free

Immunogen

The immunogen for this antibody was B7H1.

Clonality

Monoclonal

Host

Rat

Isotype

IgG2a Lambda

Scientific Data Images for PD-L1 Antibody (MIH5) - BSA Free

Flow Cytometry: PD-L1/B7-H1 Antibody (MIH5) [NBP1-43262] - B7-H1/PD-L1/CD274 Antibody (MIH5) [NBP1-43262] - Using the Allophycocyanin direct conjugate A cell surface stain was performed on RAW246.7 cells with B7-H1/PD-L1/CD274 (MIH5) antibody NBP1-43262APC (blue) and a matched isotype control NBP1-51104APC (orange). Cells were incubated in an antibody dilution of 0.5 ug/mL for 20 minutes at room temperature. Both antibodies were conjugated to Allophycocyanin.
Flow Cytometry: PD-L1/B7-H1 Antibody (MIH5) [NBP1-43262] - B7-H1/PD-L1/CD274 Antibody (MIH5) [NBP1-43262] - Analysis using the Biotin conjugate of NBP1-43262. Staining of C57Bl/6 splenocytes with 0.125 ug of Rat IgG2a Isotype Control Biotin (open histogram) or 0.125 ug of Anti-Mouse (B7-H1) Biotin (filled histogram) followed by Streptavidin PE.
Flow Cytometry: PD-L1/B7-H1 Antibody (MIH5) [NBP1-43262] - B7-H1/PD-L1/CD274 Antibody (MIH5) [NBP1-43262] - Staining of mouse splenocytes with Anti-Mouse B7-H1/PD-L1/CD274) PE. Appropriate isotype controls were used (open histogram). Total viable cells were used for analysis.

Applications for PD-L1 Antibody (MIH5) - BSA Free

Application
Recommended Usage

Flow Cytometry

1:10-1:1000

Immunohistochemistry

1:10-1:500

Immunohistochemistry-Frozen

1:10-1:500

Western Blot

1:100-1:2000
Application Notes
The MIH5 antibody has been tested by flow cytometric analysis of mouse splenocyte suspensions. This can be used at less than or equal to 0.5 ug per test. Cell number should be determined empirically but can range from 10^5 to 10^8 cells/test. Use in Immunocytochemistry/immunofluorescence reported in scientific literature (PMID: 30703170). Use In vivo reported in scientific literature (PMID: 30910830).
Please Note: Optimal dilutions of this antibody should be experimentally determined.

Reviewed Applications

Read 1 review rated 4 using NBP1-43262 in the following applications:

Published Applications

Read 9 publications using NBP1-43262 in the following applications:

Formulation, Preparation, and Storage

Purification

Protein A or G purified

Formulation

PBS (pH 7.2)

Format

BSA Free

Preservative

0.09% Sodium Azide

Concentration

0.5 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. Do not freeze.

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, anti-PD-L1, PD-L1 blocking, PD-L1 ihc, PD-L1 immunohistochemistry, PD-L1 western blot

Gene Symbol

CD274

Product Documents for PD-L1 Antibody (MIH5) - 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 (MIH5) - 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 (MIH5) - BSA Free

Showing  1 - 2 of 2 FAQs Showing All
  • Q: Can you recommend a mouse tissue positive control for this antibody hen used in IHC labelling?

    A: CD274 in mouse is highly expressed in the heart, thymus, skeletal muscle, and lung. Weakly expressed in the kidney, spleen, thyroid, and liver. Expressed on activated dendritic cells, B-cells and macrophages. Expressed in numerous tumor cells lines of lymphoid origin. (UniProt Q9EP73) I would recommend trying heart, thymus, skeletal muscle, or lung for IHC. Here is a publication that used this clone with IHC staining of mouse tissues for your reference as well: Tsushima et al. Eur J Immunol. 2003 Oct;33(10):2773-82

  • Q: On your website, it states that clone MIH5 binds to both human and mouse CD274. However, other companies state this clone only binds to mouse. What type of validation do you have that this binds to human CD274?

    A: This antibody was tested on human primary colon cells and the cancer cells including HT29 cell line by one of our customer, and you can see the related data on the datasheet of this product. Datasheet

Showing  1 - 2 of 2 FAQs Showing All
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