Human PD-L1 ELISA Kit, fluorescent is a single-wash 90-min Simplestep used to quantify Human PD-L1 with a sensitivity of 1.04 pg/ml. The assay uses a simple mix-wash-read protocol with just one incubation and one wash step.
- Fluorescent Sandwich ELISA - 530/570/590 nm readout : works on any standard plate reader
Application | Reactivity | Dilution info | Notes |
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Application sELISA | Reactivity Reacts | Dilution info - | Notes - |
Plays a critical role in induction and maintenance of immune tolerance to self (PubMed:11015443, PubMed:28813410, PubMed:28813417, PubMed:31399419). As a ligand for the inhibitory receptor PDCD1/PD-1, modulates the activation threshold of T-cells and limits T-cell effector response (PubMed:11015443, PubMed:28813410, PubMed:28813417, PubMed:36727298). Through a yet unknown activating receptor, may costimulate T-cell subsets that predominantly produce interleukin-10 (IL10) (PubMed:10581077). Can also act as a transcription coactivator: in response to hypoxia, translocates into the nucleus via its interaction with phosphorylated STAT3 and promotes transcription of GSDMC, leading to pyroptosis (PubMed:32929201). The PDCD1-mediated inhibitory pathway is exploited by tumors to attenuate anti-tumor immunity and escape destruction by the immune system, thereby facilitating tumor survival (PubMed:28813410, PubMed:28813417). The interaction with PDCD1/PD-1 inhibits cytotoxic T lymphocytes (CTLs) effector function (By similarity). The blockage of the PDCD1-mediated pathway results in the reversal of the exhausted T-cell phenotype and the normalization of the anti-tumor response, providing a rationale for cancer immunotherapy (By similarity).
CD274, B7H1, PDCD1L1, PDCD1LG1, PDL1, Programmed cell death 1 ligand 1, PD-L1, PDCD1 ligand 1, Programmed death ligand 1, hPD-L1, B7 homolog 1, B7-H1
Human PD-L1 ELISA Kit, fluorescent is a single-wash 90-min Simplestep used to quantify Human PD-L1 with a sensitivity of 1.04 pg/ml. The assay uses a simple mix-wash-read protocol with just one incubation and one wash step.
- Fluorescent Sandwich ELISA - 530/570/590 nm readout : works on any standard plate reader
Sample | n | mean | SD | C.V. |
---|---|---|---|---|
Sample Extract | n 8 | mean - | SD - | C.V. 2.7 |
Sample | n | mean | SD | C.V. |
---|---|---|---|---|
Sample Extract | n 3 | mean - | SD - | C.V. 4.3 |
Sample type | Average % | Range |
---|---|---|
Sample type Serum | Average % = 97 | Range 94 - 97 % |
Sample type EDTA Plasma | Average % = 90 | Range 89 - 92 % |
Sample type Cell culture extracts | Average % = 102 | Range 99 - 106 % |
Sample type Urine | Average % = 93 | Range 86 - 97 % |
Sample type Cell culture media | Average % = 93 | Range 82 - 100 % |
Sample type Heparin Plasma | Average % = 101 | Range 90 - 111 % |
Sample type Citrate plasma | Average % = 90 | Range 87 - 95 % |
Human PD-L1 in vitro CatchPoint® SimpleStep ELISA® (Enzyme-Linked Immunosorbent Assay) kit is designed for the quantitative measurement of PD-L1 protein in human serum, plasma, urine, cell extract and cell culture supernatants.
This CatchPoint SimpleStep ELISA kit has been optimized for Molecular Devices Microplate Readers. Click here for a list of recommended Microplate Readers.
If using a Molecular Devices' plate reader supported by SoftMax® Pro software, a preconfigured protocol for these CatchPoint SimpleStep ELISA Kits is available with all the protocol and analysis settings at www.softmaxpro.org
The CatchPoint SimpleStep ELISA employs an affinity tag labeled capture antibody and a reporter conjugated detector antibody which immunocapture the sample analyte in solution. This entire complex (capture antibody/analyte/detector antibody) is in turn immobilized via immunoaffinity of an anti-tag antibody coating the well. To perform the assay, samples or standards are added to the wells, followed by the antibody mix. After incubation, the wells are washed to remove unbound material. CatchPoint HRP Development Solution containing the Stoplight Red Substrate is added. During incubation, the substrate is catalyzed by HRP generating a fluorescent product. Signal is generated proportionally to the amount of bound analyte and the intensity is measured in a fluorescence plater reader at 530/570/590 nm Excitation/Cutoff/Emission.
PD-L1 (also known as CD274 or B7-H1) is a membrane bound glycoprotein involved in regulation of the immune system. PD-L1 is expressed on a variety of inflammatory-activated cells as well as some carcinomas and in melanoma. PD-L1 binds to PD-1 and CD80, where it can suppress T cell activation and proliferation as well as induce apoptosis. Levels of PD-L1 are increased in the plasma of cancer patients as well as in cerebrospinal fluid of gliomas. PD-L1 can bind PD-1 in order to regulate T cell apoptosis.
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PD-L1 also known as Programmed Death-Ligand 1 or CD274 is a protein involved in immune modulation. Mechanically PD-L1 interacts with its receptors particularly PD-1 to regulate cellular immune responses. This transmembrane protein has a calculated molecular weight of approximately 33 kDa. PD-L1 is expressed on various cell types including tumor cells and immune cells such as dendritic cells macrophages and B cells. Its expression is often upregulated in response to inflammatory cytokines.
PD-L1 plays a central role in immune evasion mechanisms utilized by tumors. It is not part of a larger protein complex but directly interacts with PD-1 and CD80. When PD-L1 binds to PD-1 it sends inhibitory signals leading to decreased T cell activation and proliferation allowing cancer cells to avoid immune destruction. PD-L1 expression provides a mechanism for tumors to suppress immune surveillance facilitating tumor progression.
PD-L1 is integral to the immune checkpoint pathway which is an important regulator of immune response. The interaction between PD-L1 and PD-1 provides a mechanism for immune tolerance which is part of the broader adaptive immune system pathway. PD-L1 is related to other immune checkpoint proteins such as CTLA-4 in its function to limit autoreactivity and promote immune homeostasis.
PD-L1 is most associated with cancer particularly in tumors such as melanoma and non-small cell lung cancer. PD-L1 expression on tumor cells often correlates with poor prognosis. PD-L1 directly interacts with PD-1 in these cancers a target for immunotherapies such as checkpoint inhibitors which aim to block this interaction and restore immune activity against tumors. PD-L1 involvement extends to autoimmune disorders where altered expression can impact tolerance and lead to immune-related tissue damage.
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Example of human PD-L1 standard curve in Sample Diluent NS.
The PD-L1 standard curve was prepared as described in Section 10. Raw data generated on SpectraMax iD3 Multi-Mode Microplate Reader is shown in the table. Background-subtracted data values (mean +/- SD) are graphed.
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