VEGFA ELISA Kit

Catalog No. ABIN6975430

Product Details VEGFA ELISA Kit

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

Reactivity: Human

Detection Method: Colorimetric

Method Type: Sandwich ELISA

Detection Range: 31.25 pg/mL - 2000 pg/mL

Minimum Detection Limit: 31.25 pg/mL

Application: ELISA

Purpose: For the quantitative determination of human vascular endothelial cell growth factor (VEGF) concentrations in serum, plasma, cell culture supernates, tissue homogenates, cell lysates, urine.

Sample Type: Cell Culture Supernatant, Cell Lysate, Plasma, Serum, Tissue Homogenate, Urine

Analytical Method: Quantitative

Specificity: This assay has high sensitivity and excellent specificity for detection of human VEGF. No significant cross-reactivity or interference between human VEGF and analogues was observed. Note: Limited by current skills and knowledge, it is impossible for us to complete the cross-reactivity detection between human VEGF and all the analogues, therefore, cross reaction may still exist.

Sensitivity: 25.297 pg/mL

Components:

• Assay plate
• Standard
• HRP-avidin (100 x concentrate)
• Biotin-antibody (100 x concentrate)
• Sample Diluent
• HRP-avidin Diluent
• Biotin-antibody Diluent
• Wash Buffer (25 x concentrate)
• TMB Substrate
• Stop Solution
• Adhesive Strip

Application Details

Application Notes: Optimal working dilution should be determined by the investigator.

Sample Volume: 100 μL

Assay Time: 1 - 4.5 h

Plate: Pre-coated

Protocol:

1. Prepare reagents, samples and standards as instructed.
2. Add 100 µL standard or sample to each well. Incubate 2 hours at 37 °C.
3. Remove the liquid of each well, don't wash.
4. Add 100 µL Biotin-antibody (1x) to each well. Incubate 1 hour at 37 °C.
5. Aspirate and wash 3 times.
6. Add 100 µL HRP-avidin (1x) to each well. Incubate 1 hour at 37 °C
7. Aspirate and wash 5 times.
8. Add 90 μL of TMB Substrate to each well. Incubate for 15-30 minutes at 37 °C. Protect from light.
9. Add 50 µL Stop Solution to each well. Read at 450 nm within 5 minutes.

Reagent Preparation:

1. Biotin-antibody (1x) - Centrifuge the vial before opening. Biotin-antibody requires a 100-fold dilution. A suggested 100-fold dilution is 10 μL of Biotin-antibody + 990 μL of Biotin-antibody Diluent.
2. HRP-avidin (1x) - Centrifuge the vial before opening. HRP-avidin requires a 100-fold dilution. A suggested 100-fold dilution is 10 μL of HRP-avidin + 990 μL of HRP-avidin Diluent.
3. Wash Buffer (1x) - If crystals have formed in the concentrate, warm up to room temperature and mix gently until the crystals have completely dissolved. Dilute 20 mL of Wash Buffer Concentrate (25 x) into deionized or distilled water to prepare 500 mL of Wash Buffer (1 x).
4. Standard Centrifuge the standard vial at 6000-10000rpm for 30s. Reconstitute the Standard with 1.0 mL of Sample Diluent. Do not substitute other diluents. This reconstitution produces a stock solution of 2000 pg/mL. Mix the standard to ensure complete reconstitution and allow the standard to sit for a minimum of 15 minutes with gentle agitation prior to making dilutions. Pipette 250 μL of Sample Diluent into each tube (S0-S6). Use the stock solution to produce a 2-fold dilution series (below). Mix each tube thoroughly before the next transfer. The undiluted Standard serves as the high standard (2000 pg/mL). Sample Diluent serves as the zero standard (0 pg/mL).

Note:

• Kindly use graduated containers to prepare the reagent. Please don't prepare the reagent directly in the Diluent vials provided in the kit.
• Bring all reagents to room temperature (18-25 °C) before use for 30 min.
• Prepare fresh standard for each assay. Use within 4 hours and discard after use.
• Making serial dilution in the wells directly is not permitted.
• Please carefully reconstitute Standards according to the instruction, and avoid foaming and mix gently until the crystals have completely dissolved. To minimize imprecision caused by pipetting, use small volumes and ensure that pipettors are calibrated. It is recommended to suck more than 10 μL for once pipetting.
• Distilled water is recommended to be used to make the preparation for reagents. 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): CV%<8% Three samples of known concentration were tested twenty times on one plate to assess.
Inter-assay Precision (Precision between assays): CV%<10% Three samples of known concentration were tested in twenty assays to assess.

Restrictions: For Research Use only

Handling

Storage: 4 °C,-20 °C

Storage Comment: Unopened kit Store at 2 - 8°C. Do not use the kit beyond the expiration date. May be stored for up to 1 month at 2 - 8°C. Coated assay Try to keep it in a sealed aluminum foil bag, plate and avoid the damp. Standard May be stored for up to 1 month at 2 - 8° C. If Biotin-antibody don't make recent use, better keep it store at HRP-avidin -20°C. Biotin-antibody Diluent Opened kit HRP-avidin Diluent Sample May be stored for up to 1 month at 2 - 8°C. Diluent Wash Buffer TMB Substrate Stop Solution *Provided this is within the expiration date of the kit.

Expiry Date: 6 months

References for VEGFA ELISA Kit (ABIN6975430)

You, Bi, Li, Zhang, Chen, Zhang, Li: "IL-17 induces reactive astrocytes and up-regulation of vascular endothelial growth factor (VEGF) through JAK/STAT signaling." in: Scientific reports, Vol. 7, pp. 41779, (2018) (PubMed).

Yu, Lu, Li, Zhou, Zeng, Zhan, Yuan, Yang, Xia: "Correlation of Plasma Vascular Endothelial Growth Factor and Endostatin Levels with Symptomatic Intra- and Extracranial Atherosclerotic Stenosis in a Chinese Han Population." in: Journal of stroke and cerebrovascular diseases : the official journal of National Stroke Association, Vol. 26, Issue 5, pp. 1061-1070, (2017) (PubMed).

Vingolo, Fragiotta, Mafrici, Cutini, Marinelli, Concistrè, Iannucci, Petramala, Letizia: "Vitreous and plasma changes of endothelin-1, adrenomedullin and vascular endothelium growth factor in patients with proliferative diabetic retinopathy." in: European review for medical and pharmacological sciences, Vol. 21, Issue 4, pp. 662-668, (2017) (PubMed).

Asadi, Torkaman, Hedayati, Mohajeri-Tehrani, Ahmadi, Gohardani: "Angiogenic effects of low-intensity cathodal direct current on ischemic diabetic foot ulcers: A randomized controlled trial." in: Diabetes research and clinical practice, Vol. 127, pp. 147-155, (2017) (PubMed).

Cui, Joo, Lee, Yu, Jeong, Kim, Seo: "Low temperature plasma induces angiogenic growth factor via up-regulating hypoxia-inducible factor 1α in human dermal fibroblasts." in: Archives of biochemistry and biophysics, Vol. 630, pp. 9-17, (2017) (PubMed).

Huang, Wang, Xu, Wang, Yin, Lai, Liao, Hou, Hu: "Exercise training with dietary restriction enhances circulating irisin level associated with increasing endothelial progenitor cell number in obese adults: an intervention study." in: PeerJ, Vol. 5, pp. e3669, (2017) (PubMed).

Nie, Wu, Zhu, Zhang, Zheng, Fang, You, Zhong, Xia, Hong: "Angiogenic factors are associated with development of acute graft-versus-host disease after allogeneic hematopoietic stem cell transplantation." in: Journal of Huazhong University of Science and Technology. Medical sciences = Hua zhong ke ji da xue xue bao. Yi xue Ying De wen ban = Huazhong keji daxue xuebao. Yixue Yingdewen ban, Vol. 35, Issue 5, pp. 694-699, (2016) (PubMed).

Shahidi-Dadras, Haghighatkhah, Abdollahimajd, Younespour, Partovi Kia, Zargari: "Correlation between vascular endothelial growth factor and subclinical atherosclerosis in patients with psoriasis." in: International journal of dermatology, (2015) (PubMed).

Ambreen, Ismail, Qureshi: "Association of gene polymorphism with serum levels of inflammatory and angiogenic factors in Pakistani patients with age-related macular degeneration." in: Molecular vision, Vol. 21, pp. 985-99, (2015) (PubMed).

Ayada, Toru, Genç, Akc?lar, ?ahin: "Balanced oxidative status by nesfatin-1 in intestinal ischemia-reperfusion." in: International journal of clinical and experimental medicine, Vol. 8, Issue 3, pp. 3318-24, (2015) (PubMed).

Yang, Zhang, Lu, Jia, Liang, Dong, Ye, Qin: "MicroRNA-26a suppresses angiogenesis in human hepatocellular carcinoma by targeting hepatocyte growth factor-cMet pathway." in: Hepatology (Baltimore, Md.), Vol. 59, Issue 5, pp. 1874-85, (2014) (PubMed).

Guo, Jiang, Tang, Si, Jiao: "The association of serum vascular endothelial growth factor and ferritin in diabetic microvascular disease." in: Diabetes technology & therapeutics, Vol. 16, Issue 4, pp. 224-34, (2014) (PubMed).

Umar, Asmawi, Sadikun, Majid, Al-Suede, Hassan, Altaf, Ahamed: "Ethyl-p-methoxycinnamate isolated from Kaempferia galanga inhibits inflammation by suppressing interleukin-1, tumor necrosis factor-?, and angiogenesis by blocking endothelial functions." in: Clinics (São Paulo, Brazil), Vol. 69, Issue 2, pp. 134-44, (2014) (PubMed).

Liu, Yang, Zhang, Shui, Song, Yao, Dai, Sun: "Fructopyrano-(1?4)-glucopyranose inhibits the proliferation of liver cancer cells and angiogenesis in a VEGF/VEGFR dependent manner." in: International journal of clinical and experimental medicine, Vol. 7, Issue 11, pp. 3859-69, (2014) (PubMed).

Tang, Jiao, Chang, Huang: "Risk factors for the evaluation of potential central nervous system metastasis in Burkitt's lymphoma: a case study and literature review." in: Hematological oncology, (2014) (PubMed).

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Yuan, Jin, Wei, Liu, Xue, Wang et al.: "Propranolol given orally for proliferating infantile haemangiomas: analysis of efficacy and serological changes in vascular endothelial growth factor and endothelial nitric oxide synthase in 35 ..." in: The British journal of oral & maxillofacial surgery, Vol. 51, Issue 7, pp. 656-61, (2013) (PubMed).

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Target Details for VEGFA

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

Alternative Name: vascular endothelial growth factor A (VEGFA Products)

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

Background:

Abbreviation: VEGFA

Alias: RP1-261G23.1, MGC70609, MVCD1, VEGF, VEGF-A, VPF, vascular endothelial growth factor isoform VEGF165|vascular permeability factor

UniProt: P15692

Pathways: RTK Signaling, 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