CD63 ELISA Kit

Catalog No. ABIN6962593

Product Details CD63 ELISA Kit

Target: CD63

Reactivity: Human

Detection Method: Colorimetric

Method Type: Sandwich ELISA

Detection Range: 0.16 ng/mL - 10 ng/mL

Minimum Detection Limit: 0.16 ng/mL

Application: ELISA

Purpose: The kit is a sandwich enzyme immunoassay technique for the in vitro quantitative measurement in various sample types.

Sample Type: Cell Culture Supernatant, Plasma, Serum

Analytical Method: Quantitative

Specificity: This kit recognizes Human TSPAN30/CD63 in samples. No Significant cross-reactivity or interference between Human TSPAN30/CD63 and analogues was observed.

Sensitivity: 0.1 ng/mL

Components:

• Pre-coated, ready to use 96-well strip plate, flat buttom
• Plate sealer for 96 wells
• Reference Standard
• Reference Standard & Sample Diluent
• Biotinylated Detection Antibody (100 x concentrate)
• HRP Conjugate (100 x concentrate)
• Biotinylated Detection Antibody Diluent
• HRP Conjugate Diluent
• Substrate Reagent
• Stop Solution
• Wash Buffer (25 x concentrate)
• Instruction manual

Application Details

Sample Volume: 100 μL

Assay Time: 3.5 h

Plate: Pre-coated

Protocol:

1. Add 100 µL standard or sample to each well. Incubate for 90 min at 37 °C.
2. Remove the liquid. Add 100 µL Biotinylated Detection Antibody. Incubate for 1 hour at 37 °C.
3. Aspirate and wash 3 times.
4. Add 100 µL HRP Conjugate. Incubate for 30 min at 37 °C.
5. Aspirate and wash 5 times.
6. Add 90 µL Substrate Reagent. Incubate for 15 min at 37 °C.
7. Add 50 µL Stop Solution. Read at 450 nm immediately.
8. Calculation of results.

Reagent Preparation:

1. Bring all reagents to room temperature (18~25 °C) before use. Follow the Microplate reader manual for set-up and preheat it for 15 min before OD measurement.
2. Wash Buffer: Dilute 30 mL of Concentrated Wash Buffer with 720 mL of deionized or distilled water to prepare 750 mL of Wash Buffer.Note: if crystals have formed in the concentrate, warm it in a 40 °C water bath and mix it gently until the crystals have completely dissolved
3. Standard working solution: Centrifuge the standard at 10,000xg for 1 min. Add 1.0 mL of Reference Standard &Sample Diluent, let it stand for 10 min and invert it gently several times. After it dissolves fully, mix it thoroughly with a pipette. This reconstitution produces a working solution of 10 ng/mL. Then make serial dilutions as needed. The recommended dilution gradient is as follows: 10, 5, 2.5, 1.25, 0.63, 0.32, 0.16, 0 ng/mL. Dilution method: Take 7 EP tubes, add 500 μLof Reference Standard & Sample Diluent to each tube. Pipette 500 μLof the 10 ng/mL working solution to the first tube and mix up to produce a 5 ng/mL working solution. Pipette 500 μLof the solution from the former tube into the latter one according to these steps. The illustration below is for reference. Note: the last tube is regarded as a blank. Don't pipette solution into it from the former tube.
4. Biotinylated Detection Antibody working solution: Calculate the required amount before the experiment (100 μL/well). In preparation, slightly more than calculated should be prepared. Centrifuge the stock tube before use, dilute the 100x Concentrated Biotinylated Detection Antibody to 1xworking solution with Biotinylated Detection Antibody Diluent.
5. Concentrated HRP Conjugate working solution: Calculate the required amount before the experiment (100 μL/well). In preparation, slightly more than calculated should be prepared. Dilute the 100x Concentrated HRP Conjugate to 1x working solution with Concentrated HRP Conjugate Diluent.

Sample Preparation:

• It is recommended to use fresh samples without long storage, otherwise protein degradation and denaturation may occur in these samples, leading to false results. Samples should therefore be stored for a short period at 2 - 8 °C or aliquoted at -20 °C (≤1 month) or -80 °C (≤ 3 months). Repeated freeze-thaw cycles should be avoided. Prior to assay, the frozen samples should be slowly thawed and centrifuged to remove precipitates.
• If the sample type is not specified in the instructions, a preliminary test is necessary to determine compatibility with the kit.
• If a lysis buffer is used to prepare tissue homogenates or cell culture supernatant, there is a possibility of 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 the range of the standard curve, the optimal sample dilution for the particular experiment has to be determined.

Assay Precision: Intra-assay Precision (Precision within an assay): 3 samples with low, mid range and high level Human TSPAN30/CD63 were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, mid range and high level Human TSPAN30/CD63 were tested on 3 different plates, 20 replicates in each plate.
Both intra-CV and inter-CV are < 10 %.

Restrictions: For Research Use only

Handling

Storage: 4 °C,-20 °C

Storage Comment:

1. For unopened kit: All reagents should be stored according to the labels on the vials, so they are stable up to 12 months after receipt of the kit. The Reference Standard, Biotinylated Detection Antibody, HRP Conjugate and the 96-well stripe plate should be stored at -20 °C upon receipt while the other reagents should be stored at 4 °C.
2. For used kit: When the kit is used, the remaining reagents need to be stored according to the above storage condition. Besides, please return the unused wells to the foil pouch containing the desiccant pack, and zip-seal the foil pouch.

Expiry Date: 12 months

References for CD63 ELISA Kit (ABIN6962593)

Li, Liu, Zang, Zhou, Zhang, Sun, Qi, Li, Kong, Jin, Yang, Luo, Lu, Lin, Niu, Liu: "Small extracellular vesicle-bound vascular endothelial growth factor secreted by carcinoma-associated fibroblasts promotes angiogenesis in a bevacizumab-resistant manner." in: Cancer letters, Vol. 492, pp. 71-83, (2021) (PubMed).

Shi, Qu, Lin, Xie, Tu, Liu, Zhou, Cao, Li, Liu: "Deep-Fried Atractylodis Rhizoma Protects against Spleen Deficiency-Induced Diarrhea through Regulating Intestinal Inflammatory Response and Gut Microbiota." in: International journal of molecular sciences, Vol. 21, Issue 1, (2020) (PubMed).

Chigurupati, Auddy, Biyani, Chakrabarti, Stohs: "Prevention of alcohol-induced DNA damage by a proprietary glycyrrhizin/D-mannitol product: A randomized, placebo-controlled, cross-over human study." in: Alcohol (Fayetteville, N.Y.), Vol. 69, pp. 33-39, (2019) (PubMed).

Bertram, Brixius, Brinkmann: "Exercise for the diabetic brain: how physical training may help prevent dementia and Alzheimer's disease in T2DM patients." in: Endocrine, Vol. 53, Issue 2, pp. 350-63, (2017) (PubMed).

Abo-Youssef: "Protective effect of rosiglitazone, quercetin, and their combination on fructose-induced metabolic syndrome in rats." in: Indian journal of pharmacology, Vol. 47, Issue 6, pp. 620-6, (2016) (PubMed).

"36th International Symposium on Intensive Care and Emergency Medicine : Brussels, Belgium. 15-18 March 2016." in: Critical care, Vol. 20, Issue Suppl 2, pp. 94, (2016) (PubMed).

Lopes, Silveira, Camargo, Barberato-Filho, Del Fiol, Osorio-de-Castro: "Biological drugs for the treatment of psoriasis in a public health system." in: Revista de saude publica, Vol. 48, Issue 4, pp. 651-61; discussion 661, (2015) (PubMed).

Zhao, Xu, Zhang, Ji: "Cardioprotective effect of carvedilol: inhibition of apoptosis in H9c2 cardiomyocytes via the TLR4/NF-κB pathway following ischemia/reperfusion injury." in: Experimental and therapeutic medicine, Vol. 8, Issue 4, pp. 1092-1096, (2014) (PubMed).

Akai, Onai, Morishita, Mino, Shijuku, Maruyama, Arai, Itoh, Hazama, Checchetto, Szabò, Yukutake, Suematsu, Yasui, Ishiura, Uozumi: "Aquaporin AqpZ is involved in cell volume regulation and sensitivity to osmotic stress in Synechocystis sp. strain PCC 6803." in: Journal of bacteriology, Vol. 194, Issue 24, pp. 6828-36, (2013) (PubMed).

Figueras-Aloy, Salvia-Roiges, Rodriguez-Miguélez, Miracle-Echegoyen, Botet-Mussons, Marín-Soria, Carbonell-Estrany: "Impact of chorioamnionitis on exhaled nitric oxide and endotracheal aspirate levels of nitrites-nitrates and interleukin-8 in mechanically ventilated preterm neonates." in: Pediatric pulmonology, Vol. 46, Issue 6, pp. 595-603, (2011) (PubMed).

Ghaffariyeh, Honarpisheh, Heidari, Puyan, Abasov: "Brain-derived neurotrophic factor as a biomarker in primary open-angle glaucoma." in: Optometry and vision science : official publication of the American Academy of Optometry, Vol. 88, Issue 1, pp. 80-5, (2011) (PubMed).

Escobar, Cheng, Moore, Johnson, Tannahill, Baker, Moldawer, Banerjee: "Stored packed red blood cell transfusion up-regulates inflammatory gene expression in circulating leukocytes." in: Annals of surgery, Vol. 246, Issue 1, pp. 129-34, (2007) (PubMed).

Target Details for CD63

Target: CD63

Alternative Name: Tetraspanin 30 (CD63 Products)

Synonyms: C75951 ELISA Kit, ME491 ELISA Kit, Tspan30 ELISA Kit, LAMP-3 ELISA Kit, MLA1 ELISA Kit, OMA81H ELISA Kit, TSPAN30 ELISA Kit, TSP29FA ELISA Kit, CD63 antigen ELISA Kit, CD63 molecule ELISA Kit, Cd63 molecule ELISA Kit, Cd63 ELISA Kit, CD63 ELISA Kit