VEGFA Kit ELISA

N° du produit ABIN6963448

Détail du produit VEGFA Kit ELISA

Antigène: VEGFA (Vascular Endothelial Growth Factor A (VEGFA))

Reactivité: Humain

Méthode de détection: Colorimetric

Type de méthode: Sandwich ELISA

Gamme de detection: 31.25 pg/mL - 2000 pg/mL

Seuil minimal de détection: 31.25 pg/mL

Application: ELISA

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

Type d'échantillon: Cell Culture Supernatant, Plasma, Serum

Analytical Method: Quantitative

Specificité: This kit recognizes Human VEGF-A in samples. No Significant cross-reactivity or interference between Human VEGF-A and analogues was observed.

Sensibilité: 18.75 pg/mL

Ingrédients:

• 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

Information d'application

Volume d'échantillon: 100 μL

Durée du test: 3.5 h

Plaque: Pre-coated

Protocole:

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.

Préparation des réactifs:

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 2000 pg/mL. Then make serial dilutions as needed. The recommended dilution gradient is as follows: 2000, 1000, 500, 250, 125, 62.5, 31.25, 0 pg/mL. Dilution method: Take 7 EP tubes, add 500 μLof Reference Standard & Sample Diluent to each tube. Pipette 500 μLof the 2000 pg/mL working solution to the first tube and mix up to produce a 1000 pg/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.

Préparation de l'échantillon:

• 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.

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

Restrictions: For Research Use only

Stockage

Stock: 4 °C,-20 °C

Stockage commentaire:

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.

Date de péremption: 12 months

Références pour VEGFA Kit ELISA (ABIN6963448)

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." dans: Cancer letters, Vol. 492, pp. 71-83, (2021) (PubMed).

Yufei, Yuqi, Binyue, Lingchen, Xi, Hoffelt, Fuliang: "Chrysin Inhibits Melanoma Tumor Metastasis via Interfering with the FOXM1/β-Catenin Signaling." dans: Journal of agricultural and food chemistry, Vol. 68, Issue 35, pp. 9358-9367, (2021) (PubMed).

Skrypnik, Mostowska, Jagodziński, Bogdański et al.: "Association of rs699947 (-2578 C/A) and rs2010963 (-634 G/C) Single Nucleotide Polymorphisms of the VEGF Gene, VEGF-A and Leptin Serum Level, and Cardiovascular Risk in Patients with Excess Body ..." dans: Journal of clinical medicine, Vol. 9, Issue 2, (2020) (PubMed).

Vimalraj, Saravanan, Raghunandhakumar, Anuradha: "Melatonin regulates tumor angiogenesis via miR-424-5p/VEGFA signaling pathway in osteosarcoma." dans: Life sciences, Vol. 256, pp. 118011, (2020) (PubMed).

Liu, Xu, Su, Fan: "REC8 suppresses tumor angiogenesis by inhibition of NF-κB-mediated vascular endothelial growth factor expression in gastric cancer cells." dans: Biological research, Vol. 53, Issue 1, pp. 41, (2020) (PubMed).

Li, Fang, Zhang, Wang, Li, Yan, Yu, Cheng, Sun: "Melatonin stimulates VEGF expression in human granulosa-lutein cells: A potential mechanism for the pathogenesis of ovarian hyperstimulation syndrome." dans: Molecular and cellular endocrinology, Vol. 518, pp. 110981, (2020) (PubMed).

Khan, Paterson, Roshandel, Raza, Ajmal, Waheed, Azam, Qamar: "Association of IGF1 and VEGFA polymorphisms with diabetic retinopathy in Pakistani population." dans: Acta diabetologica, Vol. 57, Issue 2, pp. 237-245, (2020) (PubMed).

Yueniwati, Darmiastini, Arisetijono: "Thicker carotid intima-media thickness and increased plasma VEGF levels suffered by post-acute thrombotic stroke patients." dans: International journal of general medicine, Vol. 9, pp. 447-452, (2016) (PubMed).

Détail du antigène

Antigène: VEGFA (Vascular Endothelial Growth Factor A (VEGFA))

Autre désignation: Vascular Endothelial Cell Growth Factor A (VEGFA Produits)

Synonymes: MVCD1 Kit ELISA, VEGF Kit ELISA, VPF Kit ELISA, Vegf Kit ELISA, Vegf120 Kit ELISA, Vegf164 Kit ELISA, Vegf188 Kit ELISA, Vpf Kit ELISA, vegf Kit ELISA, vegfa Kit ELISA, wu:fj82c06 Kit ELISA, VEGF-A Kit ELISA, VEGF164 Kit ELISA, eVEGF120 Kit ELISA, eVEGF164 Kit ELISA, vegf-a Kit ELISA, vpf Kit ELISA, vefg Kit ELISA, si:dkey-14d8.3 Kit ELISA, wu:fd42e02 Kit ELISA, vascular endothelial growth factor A Kit ELISA, vascular endothelial growth factor Aa Kit ELISA, vascular endothelial growth factor A L homeolog Kit ELISA, vascular endothelial growth factor Ab Kit ELISA, VEGFA Kit ELISA, Vegfa Kit ELISA, vegfaa Kit ELISA, vegfa.L Kit ELISA, vegfa Kit ELISA, vegfab Kit ELISA

Sujet: VEGF-A, VEGFA, MVCD1, VPF

Pathways: Signalisation RTK, Glycosaminoglycan Metabolic Process, Regulation of Cell Size, Tube Formation, Signaling Events mediated by VEGFR1 and VEGFR2, Platelet-derived growth Factor Receptor Signaling, VEGFR1 Specific Signals, VEGF Signaling