PD-L1 ELISA Kit

Catalog No. ABIN6574208

Product Details PD-L1 ELISA Kit

Target: PD-L1 (CD274 (PD-L1))

Reactivity: Human

Detection Method: Colorimetric

Method Type: Sandwich ELISA

Detection Range: 0.15 ng/mL - 10 ng/mL

Minimum Detection Limit: 0.15 ng/mL

Application: ELISA

Purpose: The kit is a sandwich enzyme immunoassay for in vitro quantitative measurement of PDCD1LG1 in human serum, plasma, tissue homogenates.

We offer validation data (WB) for the kit components. So you can be sure to order a reliable ELISA kit product composed of high quality reagents.

Sample Type: Plasma, Serum, Tissue Homogenate

Analytical Method: Quantitative

Specificity: This assay has high sensitivity and excellent specificity for detection of Programmed Cell Death Protein 1 Ligand 1 (PDL1)

Cross-Reactivity (Details): No significant cross-reactivity or interference between Programmed Cell Death Protein 1 Ligand 1 (PDCD1LG1) and analogues was observed.

Sensitivity: 0.056 ng/mL

Components:

• Pre-coated, ready to use 96-well strip plate, flat buttom
• Plate sealer for 96 wells
• Reference Standard
• Standard Diluent
• Detection Reagent A
• Detection Reagent B
• Assay Diluent A
• Assay Diluent B
• Reagent Diluent (if Detection Reagent is lyophilized)
• TMB Substrate
• Stop Solution
• Wash Buffer (30 x concentrate)
• Instruction manual

Application Details

Comment:

Information on standard material:
The standard might be recombinant protein or natural protein, that will depend on the specific kit. Moreover, the expression system is E.coli or yeast or mammal cell. There is 0.05% proclin 300 in the standard as preservative.

Information on reagents:
The stop solution used in the kit is sulfuric acid with concentration of 1 mol/L. And the wash solution is TBS. The standard diluent contains 0.02 % sodium azide, assay diluent A and assay diluent B contain 0.01% sodium azide. Some kits can contain is BSA in them.

Information on antibodies:
The provided antibodies and their host vary in different kits.

Sample Volume: 100 μL

Assay Time: 3 h

Plate: Pre-coated

Protocol:

1. Prepare all reagents, samples and standards,
2. Add 100μL standard or sample to each well. Incubate 1 hours at 37 °C,
3. Aspirate and add 100μL prepared Detection Reagent A. Incubate 1 hour at 37 °C,
4. Aspirate and wash 3 times,
5. Add 100μL prepared Detection Reagent B. Incubate 30 minutes at 37 °C,
6. Aspirate and wash 5 times,
7. Add 90μL Substrate Solution. Incubate 10-20 minutes at 37 °C,
8. Add 50μL Stop Solution. Read at 450nm immediately.

Reagent Preparation:

1. Bring all kit components and samples to room temperature (18-25 °C) before use. If the kit will not be used up in one time, please only take out strips and reagents for present experiment, and leave the remaining strips and reagents in required condition.
2. Standard - Reconstitute the Standard with 1.0 mL of Standard Diluent, keep for 10 minutes at room temperature, shake gently (not to foam). The concentration of the standard in the stock solution is 40 ng/mL. First dilute the stock solution to 10 ng/mL and the diluted standard serves as the highest standard (10 ng/mL). Then prepare 7 tubes containing 0.5 mL Standard Diluent and use the diluted standard to produce a double dilution series. Mix each tube thoroughly before the next transfer. Set up 7 points of diluted standard such as 10 ng/mL, 5 ng/mL, 2.5 ng/mL, 1.25 ng/mL, 0.625 ng/mL, 0.312 ng/mL, 0.156 ng/mL, and the last microcentrifuge tube with Standard Diluent is the blank as 0 ng/mL.
3. Detection Reagent A and Detection Reagent B - If lyophilized reconstitute the Detection Reagent A with 150μL of Reagent Diluent, keep for 10 minutes at room temperature, shake gently (not to foam). Briefly spin or centrifuge the stock Detection A and Detection B before use. Dilute them to the working concentration 100-fold with Assay Diluent A and B, respectively.
4. Wash Solution - Dilute 20 mL of Wash Solution concentrate (30x) with 580 mL of deionized or distilled water to prepare 600 mL of Wash Solution (1x).
5. TMB substrate - Aspirate the needed dosage of the solution with sterilized tips and do not dump the residual solution into the vial again.

Note:

1. Making serial dilution in the wells directly is not permitted.
2. Prepare standards within 15 minutes before assay. Please do not dissolve the reagents at 37 °C directly.
3. Please carefully reconstitute Standards or working Detection Reagent A and B according to the instruction, and avoid foaming and mix gently until the crystals are completely dissolved. To minimize imprecision caused by pipetting, use small volumes and ensure that pipettors are calibrated. It is recommended to suck more than 10μL for one pipetting.
4. The reconstituted Standards, Detection Reagent A and Detection Reagent B can be used only once.
5. If crystals have formed in the Wash Solution concentrate (30x), warm to room temperature and mix gently until the crystals are completely dissolved.
6. Contaminated water or container for reagent preparation will influence the detection result.

Sample Preparation:

• It is recommended to use fresh samples without long storage, otherwise protein degradation and denaturationmay occur in these samples, leading to false results. Samples should therefore be stored for a short periodat 2 - 8 °C or aliquoted at -20 °C (≤1 month) or -80 °C (≤ 3 months). Repeated freeze-thawcycles should be avoided. Prior to assay, the frozen samples should be slowly thawed and centrifuged toremove precipitates.
• If the sample type is not specified in the instructions, a preliminary test is necessary to determinecompatibility with the kit.
• If a lysis buffer is used to prepare tissue homogenates or cell culture supernatant, there is a possibilityof causing a deviation due to the introduced chemical substance.The recommended dilution factor is for reference only.
• Please estimate the concentration of the samples before performing the test. If the values are not in therange of the standard curve, the optimal sample dilution for the particular experiment has to be determined.Samples should then be diluted with PBS (pH =7.0-7.2).

Assay Precision: Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level of target were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level of target were tested on 3 different plates, 8 replicates in each plate.
CV(%) = SD/meanX100
Intra-Assay: CV < 10%
Inter-Assay: CV < 12%

Restrictions: For Research Use only

Handling

Precaution of Use: The Stop Solution suggested for use with this kit is an acid solution. Wear eye, hand, face, and clothing protection when using this material.

Storage: 4 °C/-20 °C

Storage Comment:

1. For unopened kit: All reagents should be stored according to the labels on the vials. The Standard, Detection Reagent A, Detection Reagent B, and 96-well Strip Plate should be stored at -20 °C upon receipt, while the other reagents should be stored at 4 °C.
2. For opened kits: the remaining reagents must be stored according to the above storage conditions. In addition, please return the unused wells to the foil pouch containing the desiccant and seal the foil pouch with the zipper.

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Expiry Date: 6 months

References for PD-L1 ELISA Kit (ABIN6574208)

Tominaga, Akiyoshi, Yamamoto, Taguchi, Mori, Nagasaki, Fukunaga, Ueno: "Clinical significance of soluble programmed cell death-1 and soluble programmed cell death-ligand 1 in patients with locally advanced rectal cancer treated with neoadjuvant chemoradiotherapy." in: PLoS ONE, Vol. 14, Issue 2, pp. e0212978, (2019) (PubMed).

Okuma, Wakui, Utsumi, Sagawa, Hosomi, Kuwano, Homma: "Soluble Programmed Cell Death Ligand 1 as a Novel Biomarker for Nivolumab Therapy for Non-Small-cell Lung Cancer." in: Clinical lung cancer, Vol. 19, Issue 5, pp. 410-417.e1, (2019) (PubMed).

Chen, Peng, Yang, Yin, Shi, Zhang, Wang: "Increased Levels of Soluble Programmed Death Ligand 1 Associate with Malignancy in Patients with Dermatomyositis." in: The Journal of rheumatology, Vol. 45, Issue 6, pp. 835-840, (2018) (PubMed).

Guo, Wang, Jin, Chen, Zhen, Jiang, Lin, Huang, Xia, Sun: "High Serum Level of Soluble Programmed Death Ligand 1 is Associated With a Poor Prognosis in Hodgkin Lymphoma." in: Translational oncology, Vol. 11, Issue 3, pp. 779-785, (2018) (PubMed).

Zhao, Zhang, Wang, Xi, Zhao, Ji, Hu: "Plasma levels of soluble programmed death ligand-1 may be associated with overall survival in nonsmall cell lung cancer patients receiving thoracic radiotherapy." in: Medicine, Vol. 96, Issue 7, pp. e6102, (2017) (PubMed).

Okuma, Hosomi, Nakahara, Watanabe, Sagawa, Homma: "High plasma levels of soluble programmed cell death ligand 1 are prognostic for reduced survival in advanced lung cancer." in: Lung cancer (Amsterdam, Netherlands), Vol. 104, pp. 1-6, (2017) (PubMed).

Nagato, Ohkuri, Ohara, Hirata, Kishibe, Komabayashi, Ueda, Takahara, Kumai, Ishibashi, Kosaka, Aoki, Oikawa, Uno, Akiyama, Sado, Takei, Celis, Harabuchi, Kobayashi: "Programmed death-ligand 1 and its soluble form are highly expressed in nasal natural killer/T-cell lymphoma: a potential rationale for immunotherapy." in: Cancer immunology, immunotherapy : CII, Vol. 66, Issue 7, pp. 877-890, (2017) (PubMed).

Finkelmeier, Canli, Tal, Pleli, Trojan, Schmidt, Kronenberger, Zeuzem, Piiper, Greten, Waidmann: "High levels of the soluble programmed death-ligand (sPD-L1) identify hepatocellular carcinoma patients with a poor prognosis." in: European journal of cancer (Oxford, England : 1990), Vol. 59, pp. 152-9, (2016) (PubMed).

Wang, Wang, Liu, Xia, Huang, Jiang, Li, Lu: "High post-treatment serum levels of soluble programmed cell death ligand 1 predict early relapse and poor prognosis in extranodal NK/T cell lymphoma patients." in: Oncotarget, Vol. 7, Issue 22, pp. 33035-45, (2016) (PubMed).

Huang, Lin, Lin, Li, Yao, Tang, Hou, Tsay, Chou, Cheng, Tien: "Soluble PD-L1: A biomarker to predict progression of autologous transplantation in patients with multiple myeloma." in: Oncotarget, Vol. 7, Issue 38, pp. 62490-62502, (2016) (PubMed).

Nasiri Kalmarzi, Fattahi, Kaviani, Ataee, Mansouri, Moradi, Yousefzade, Abbassi: "Inverse correlation of soluble programmed cell death-1 ligand-1 (sPD-L1) with eosinophil count and clinical severity in allergic rhinitis patients." in: Allergology international : official journal of the Japanese Society of Allergology, Vol. 66, Issue 2, pp. 326-331, (2016) (PubMed).

Bi, Wang, Zhang, Wang, Liu, Xia, Huang, Jiang, Zhang, Wang: "PD-L1 is upregulated by EBV-driven LMP1 through NF-κB pathway and correlates with poor prognosis in natural killer/T-cell lymphoma." in: Journal of hematology & oncology, Vol. 9, Issue 1, pp. 109, (2016) (PubMed).

Ha, Nam, Bang, Park, Kim, Lee, Han, Im, Kim, Bang, Oh: "Soluble programmed death-ligand 1 (sPDL1) and neutrophil-to-lymphocyte ratio (NLR) predicts survival in advanced biliary tract cancer patients treated with palliative chemotherapy." in: Oncotarget, Vol. 7, Issue 47, pp. 76604-76612, (2016) (PubMed).

Keane, Vari, Hertzberg, Cao, Green, Han, Seymour, Hicks, Gill, Crooks, Gould, Jones, Griffiths, Talaulikar, Jain, Tobin, Gandhi: "Ratios of T-cell immune effectors and checkpoint molecules as prognostic biomarkers in diffuse large B-cell lymphoma: a population-based study." in: The Lancet. Haematology, Vol. 2, Issue 10, pp. e445-55, (2015) (PubMed).

Rossille, Gressier, Damotte, Maucort-Boulch, Pangault, Semana, Le Gouill, Haioun, Tarte, Lamy, Milpied, Fest: "High level of soluble programmed cell death ligand 1 in blood impacts overall survival in aggressive diffuse large B-Cell lymphoma: results from a French multicenter clinical trial." in: Leukemia, Vol. 28, Issue 12, pp. 2367-75, (2014) (PubMed).

Pizarro, García-Díaz, Codner, Salas-Pérez, Carrasco, Pérez-Bravo: "PD-L1 gene polymorphisms and low serum level of PD-L1 protein are associated to type 1 diabetes in Chile." in: Diabetes/metabolism research and reviews, Vol. 30, Issue 8, pp. 761-6, (2014) (PubMed).

Zheng, Bu, Liu, Zhang, Li, Wu, Wu, Cheng, Xing, Du, Wang, Hu, Ji: "Level of circulating PD-L1 expression in patients with advanced gastric cancer and its clinical implications." in: Chinese journal of cancer research = Chung-kuo yen cheng yen chiu, Vol. 26, Issue 1, pp. 104-11, (2014) (PubMed).

Curry, Lawrence, Burden: "Ovarian sympathectomy in the golden hamster: effects on estrous cyclicity and follicular development." in: Experimental and clinical endocrinology, Vol. 86, Issue 3, pp. 284-90, (1986) (PubMed).

Target Details for PD-L1

Target: PD-L1 (CD274 (PD-L1))

Alternative Name: Programmed Cell Death Protein 1 Ligand 1 (PDL1) (PD-L1 Products)

Synonyms: B7-H ELISA Kit, B7H1 ELISA Kit, PD-L1 ELISA Kit, PDCD1L1 ELISA Kit, PDCD1LG1 ELISA Kit, PDL1 ELISA Kit, A530045L16Rik ELISA Kit, B7h1 ELISA Kit, Pdcd1l1 ELISA Kit, Pdcd1lg1 ELISA Kit, Pdl1 ELISA Kit, RGD1566211 ELISA Kit, CD274 molecule ELISA Kit, CD274 antigen ELISA Kit, programmed cell death 1 ligand 2 ELISA Kit, CD274 ELISA Kit, Cd274 ELISA Kit, PDCD1LG2 ELISA Kit

UniProt: Q9NZQ7

Pathways: Cancer Immune Checkpoints