VEGF ELISA Kit (Vascular Endothelial Growth Factor A)

Details for Product VEGFA ELISA Kit No. ABIN625218, Supplier: Log in to see
Antigen
  • vegf
  • vegfa
  • wu:fj82c06
  • VEGF
  • vegf-a
  • vpf
  • vefg
  • VEGF-A
  • VPF
  • eVEGF120
  • eVEGF164
  • MVCD1
  • Vegf
  • Vegf120
  • Vegf164
  • Vegf188
  • Vpf
  • VEGF164
  • vascular endothelial growth factor Aa
  • vascular endothelial growth factor A L homeolog
  • vascular endothelial growth factor A
  • vegfaa
  • vegfa.L
  • VEGFA
  • vegfa
  • Vegfa
Reactivity
Rat (Rattus)
Alternatives
Kits with alternative reactivity to:
29
26
26
15
13
8
8
8
7
6
3
3
2
1
Method Type
Sandwich ELISA
Detection Range
2-200 pg/mL
Minimum Detection Limit
2 pg/mL
Application
ELISA
Options
Supplier
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Supplier Product No.
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Purpose Rat VEGF-A ELISA Kit for cell culture supernatants, plasma, and serum samples.
Sample Type Plasma, Cell Culture Supernatant, Serum
Analytical Method Quantitative
Detection Method Colorimetric
Specificity This ELISA kit shows no cross-reactivity with any of the cytokines tested: rat CINC-2, CINC-3, CNTF, Fractalkine, IL-1 alpha, IL-1 beta, IL-4, IL-6, IL-10, GM-CSF, IFN-gamma, Leptin, Lix, MCP-1, MIP-3 alpha, beta- NGF, TIMP-1, TNF-alpha.
Cross-Reactivity (Details) This ELISA kit shows no cross-reactivity with any of the cytokines tested (e.g., rat CINC-2, CINC-3, CNTF, Fractalkine, IL-1alpha, IL- 1beta, IL-4, IL-6, IL-10, GM-CSF, IFN-gamma, Leptin, Lix, MCP-1, MIP-3alpha, beta- NGF, TIMP-1, TNF-alpha).
Sensitivity < 2 pg/mL
Characteristics
  • Strip plates and additional reagents allow for use in multiple experiments
  • Quantitative protein detection
  • Establishes normal range
  • The best products for confirmation of antibody array data
Components
  • Pre-Coated 96-well Strip Microplate
  • Wash Buffer
  • Stop Solution
  • Assay Diluent(s)
  • Lyophilized Standard
  • Biotinylated Detection Antibody
  • Streptavidin-Conjugated HRP
  • TMB One-Step Substrate
Material not included
  • Distilled or deionized water
  • Precision pipettes to deliver 2 μL to 1 μL volumes
  • Adjustable 1-25 μL pipettes for reagent preparation
  • 100 μL and 1 liter graduated cylinders
  • Tubes to prepare standard and sample dilutions
  • Absorbent paper
  • Microplate reader capable of measuring absorbance at 450nm
  • Log-log graph paper or computer and software for ELISA data analysis
Plasmids, Primers & others Plasmids, Primers & others VEGF products on genomics-online (e.g. as negative or positive controls)
Antigen Vascular Endothelial Growth Factor A (VEGFA)
Alternative Name VEGF-A (VEGFA ELISA Kit Abstract)
Background Vascular endothelial growth factor A (VEGF-A) (Vascular permeability factor) (VPF)
Gene ID 83785
UniProt P16612
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
Application Notes Recommended Dilution for serum and plasma samples2 fold
Sample Volume 100 μL
Plate Pre-coated
Protocol
  1. Prepare all reagents, samples and standards as instructed in the manual.
  2. Add 100 μL of standard or sample to each well.
  3. Incubate 2.5 h at RT or O/N at 4 °C.
  4. Add 100 μL of prepared biotin antibody to each well.
  5. Incubate 1 h at RT.
  6. Add 100 μL of prepared Streptavidin solution to each well.
  7. Incubate 45 min at RT.
  8. Add 100 μL of TMB One-Step Substrate Reagent to each well.
  9. Incubate 30 min at RT.
  10. Add 50 μL of Stop Solution to each well.
  11. Read at 450 nm immediately.
Reagent Preparation
  1. Bring all reagents and samples to room temperature (18 - 25 °C) before use.
    2. Sample dilution: If your samples need to be diluted, Assay Diluent A (Item D) should be used for dilution of serum/plasma samples. 1x Assay Diluent B (Item E) should be used for dilution of culture supernatants. Suggested dilution for normal serum/plasma: 2 fold. Please note that levels of the target protein may vary between different specimens. Optimal dilution factors for each sample must be determined by the investigator.
    3. Assay Diluent B (Item E) should be diluted 5-fold with deionized or distilled water before use.
    4. Preparation of standard: Briefly spin the vial of Item C and then add 400 µL Assay Diluent A (for serum/plasma samples) or 1x Assay Diluent B (for cell culture medium) into Item C vial to prepare a 50 ng/mL standard. Dissolve the powder thoroughly by a gentle mix. Add 4 µL VEGF standard from the vial of Item C, into a tube with 996.0 µL Assay Diluent A or 1x Assay Diluent B to prepare a 200 pg/mL stock standard solution. Pipette 300 µL Assay Diluent A or 1x Assay Diluent B into each tube. Use the stock standard solution to produce a dilution series . Mix each tube thoroughly before the next transfer. Assay Diluent A or 1x Assay Diluent B serves as the zero standard (0 pg/mL). 200 µL 200myl 200 µL 200 µL 200 µL 200 µL 4 µL standard +996.0 µL 200 80 32 12.8 5.12 2.05 0.82 0 pg/mL pg/mL pg/mL pg/mL pg/mL pg/mL pg/mL pg/mL
    5. If the Wash Concentrate (20x) (Item B) contains visible crystals, warm to room temperature and mix gently until dissolved. Dilute 20 ml of Wash Buffer Concentrate into deionized or distilled water to yield 400 ml of 1x Wash Buffer.
    6. Briefly spin the Detection Antibody vial (Item F) before use. Add 100 µL of 1x Assay Diluent B into the vial to prepare a detection antibody concentrate. Pipette up and down to mix gently (the concentrate can be stored at 4 °C for 5 days). The detection antibody concentrate should be diluted 80-fold with 1x Assay Diluent B and used in step 4 of Part VI Assay Procedure.
    7. Briefly spin the HRP-Streptavidin concentrate vial (Item G) before use. HRP-Streptavidin concentrate should be diluted 120-fold with 1x Assay Diluent B. For example: Briefly spin the vial (Item G) and pipette up and down to mix gently . Add 100 µL of HRP-Streptavidin concentrate into a tube with 12 m1 1x Assay Diluent B to prepare a 120-fold diluted HRP-Streptavidin solution (don't store the diluted solution for next day use). Mix well. VI. ASSAY PROCEDURE: 1. Bring all reagents and samples to room temperature (18 - 25 °C) before use. It is recommended that all standards and samples be run at least in duplicate.
    2. Add 100 µL of each standard (see Reagent Preparation step 2) and sample into appropriate wells. Cover well and incubate for 2.5 hours at room temperature or over night at 4 °C with gentle shaking.
    3. Discard the solution and wash 4 times with 1x Wash Solution. Wash by filling each well with Wash Buffer (300 myl) using a multi-channel Pipette or autowasher. Complete removal of liquid at each step is essential to good performance. After the last wash, remove any remaining Wash Buffer by aspirating or decanting. Invert the plate and blot it against clean paper towels.
    4. Add 100 µL of 1x prepared biotinylated antibody (Reagent Preparation step 6) to each well. Incubate for 1 hour at room temperature with gentle shaking.
    5. Discard the solution. Repeat the wash as in step
    6. Add 100 µL of prepared Streptavidin solution (see Reagent Preparation step 7) to each well. Incubate for 45 minutes at room temperature with gentle shaking.
    7. Discard the solution. Repeat the wash as in step
    8. Add 100 µL of TMB One-Step Substrate Reagent (Item H) to each well. Incubate for 30 minutes at room temperature in the dark with gentle shaking.
    9. Add 50 µL of Stop Solution (Item I) to each well. Read at 450 nm immediately.
Assay Procedure
  1. Bring all reagents and samples to room temperature (18 - 25 °C) before use. It is recommended that all standards and samples be run at least in duplicate.
    2. Add 100 µL of each standard (see Reagent Preparation step 2) and sample into appropriate wells. Cover well and incubate for 2.5 hours at room temperature or over night at 4 °C with gentle shaking.
    3. Discard the solution and wash 4 times with 1x Wash Solution. Wash by filling each well with Wash Buffer (300 myl) using a multi-channel Pipette or autowasher. Complete removal of liquid at each step is essential to good performance. After the last wash, remove any remaining Wash Buffer by aspirating or decanting. Invert the plate and blot it against clean paper towels.
    4. Add 100 µL of 1x prepared biotinylated antibody (Reagent Preparation step 6) to each well. Incubate for 1 hour at room temperature with gentle shaking.
    5. Discard the solution. Repeat the wash as in step
    6. Add 100 µL of prepared Streptavidin solution (see Reagent Preparation step 7) to each well. Incubate for 45 minutes at room temperature with gentle shaking.
    7. Discard the solution. Repeat the wash as in step
    8. Add 100 µL of TMB One-Step Substrate Reagent (Item H) to each well. Incubate for 30 minutes at room temperature in the dark with gentle shaking.
    9. Add 50 µL of Stop Solution (Item I) to each well. Read at 450 nm immediately.
Calculation of Results

Calculate the mean absorbance for each set of duplicate standards, controls and samples, and subtract the average zero standard optical density. Plot the standard curve on log-log graph paper or using Sigma plot software, with standard concentration on the x-axis and absorbance on the y-axis. Draw the best-fit straight line through the standard points.
Typical Data: These standard curves are for demonstration only. A standard curve must be run with each assay. Assay Diluent A Rat VEGF concentration (pg/mL) 0.1 1 10 100 1000 O D =4 50 (n m ) 0.01 0.1 1 10 Assay Diluent B Rat VEGF concentration (pg/mL) 0.1 1 10 100 1000 O D =4 50 (n m ) 0.001 0.01 0.1 1 10
Sensitivity: The minimum detectable dose of VEGF is typically less than 2 pg/mL.
Recovery: Recovery was determined by spiking various levels of Rat VEGF into Rat serum, plasma and cell culture media. Mean recoveries are as follows: Sample Type Average % Recovery Range ( %) Serum 97.43 83-103 Plasma 96.42 85-104 Cell culture media 98.26 84-103
Linearity: Sample Type Serum Plasma Cell Culture Media 1:2 Average % of Expected 96 95 96 Range ( %) 84-103 83-103 84-104 1:4 Average % of Expected 98 96 96 Range ( %) 86-104 84-104 84-103
Reproducibility: Intra-Assay: CV<10 % Inter-Assay: CV<12 %

Assay Precision Intra-Assay: CV< 10 % Inter-Assay: CV< 12 %
Restrictions For Research Use only
Handling Advice Avoid repeated freeze-thaw cycles.
Storage -20 °C
Storage Comment The entire kit may be stored at -20°C for up to 1 year from the date of shipment. Avoid repeated freeze-thaw cycles. The kit may be stored at 4°C for up to 6 months. For extended storage, it is recommended to store at -80°C.
Expiry Date 6 months
Supplier Images
ELISA image for Vascular Endothelial Growth Factor A (VEGFA) ELISA Kit (ABIN625218) Vascular Endothelial Growth Factor A (VEGFA) ELISA Kit
Product cited in: El-Mahdy, Salem, El-Sayad, El-Desouky, Zaghow: "Bone marrow mononuclear cells enhance anti-inflammatory effects of pravastatin against isoproterenol-induced myocardial infarction in rats." in: Journal of immunotoxicology, Vol. 13, Issue 3, pp. 393-402, 2017 (PubMed). (Sample species: Rat (Rattus)). Further details: Sample: Tissue lysate (Left ventricles from rats injected with various cell types or control)

Lin, Pan, Ruan, Qian, Liang, Ge, Huang: "Differential expression of HIF-1?, AQP-1, and VEGF under acute hypoxic conditions in the non-ventilated lung of a one-lung ventilation rat model." in: Life sciences, Vol. 124, pp. 50-5, 2015 (PubMed).

Kaya, Demir, Bulut, Akpolat, Turgut: "Effects of lapatinib and trastuzumab on vascular endothelial growth factor in experimental corneal neovascularization." in: Clinical & experimental ophthalmology, Vol. 43, Issue 5, pp. 449-57, 2015 (PubMed).

Ikutomi, Sahara, Nakajima, Minami, Morita, Hirata, Komuro, Nakamura, Sata: "Diverse contribution of bone marrow-derived late-outgrowth endothelial progenitor cells to vascular repair under pulmonary arterial hypertension and arterial neointimal formation." in: Journal of molecular and cellular cardiology, Vol. 86, pp. 121-35, 2015 (PubMed).

Hao, Huang, Shi, Cheng, Li, Zhou, Guo, Li, Lu, Zhu, Duan: "Pulsed electromagnetic field improves cardiac function in response to myocardial infarction." in: American journal of translational research, Vol. 6, Issue 3, pp. 281-90, 2014

Lebrun-Harris, Fiore, Tomoyasu, Ngo-Metzger: "Cigarette Smoking, Desire to Quit, and Tobacco-Related Counseling Among Patients at Adult Health Centers." in: American journal of public health, 2014 (PubMed).

Hao, Huang, Song, Shi, Cheng, Murohara, Lu, Su, Duan: "Arterial baroreflex dysfunction impairs ischemia-induced angiogenesis." in: Journal of the American Heart Association, Vol. 3, Issue 3, pp. e000804, 2014 (PubMed).

Hao, Huang, Shi, Cheng, Li, Zhou, Guo, Li, Lu, Zhu, Duan: "Pulsed electromagnetic field improves cardiac function in response to myocardial infarction." in: American journal of translational research, Vol. 6, Issue 3, pp. 281-90, 2014 (PubMed).

Oktem, Ozcan, Erdem, Karakaya, Cenksoy, Guner, Karabacak, Dursun: "The effect of captopril on endometriotic implants in a rat model." in: European journal of obstetrics, gynecology, and reproductive biology, Vol. 180, pp. 120-5, 2014 (PubMed).

Ergeno?lu, Yeniel, Erba?, Aktu?, Yildirim, Uluku?, Taskiran: "Regression of endometrial implants by resveratrol in an experimentally induced endometriosis model in rats." in: Reproductive sciences (Thousand Oaks, Calif.), Vol. 20, Issue 10, pp. 1230-6, 2013 (PubMed).

Gandhi, Srinivasan, Akarte: "Aliskiren improves insulin resistance and ameliorates diabetic renal vascular complications in STZ-induced diabetic rats." in: Journal of the renin-angiotensin-aldosterone system : JRAAS, Vol. 14, Issue 1, pp. 3-13, 2013 (PubMed).

Yüksel, Yavuz, I?, Çomuno?lu, Üzüm, Akyüz, Y?ld?r?m: "Simvastatin reduces VEGF and NO levels in acute stages of experimental traumatic brain injury." in: Neurological sciences : official journal of the Italian Neurological Society and of the Italian Society of Clinical Neurophysiology, Vol. 34, Issue 11, pp. 1941-6, 2013 (PubMed).

Maillard, Juszczak, Langlois, Kleiss, Sencier, Bietiger, Sanchez-Dominguez, Krafft, Johnson, Pinget, Sigrist: "Perfluorocarbon emulsions prevent hypoxia of pancreatic ?-cells." in: Cell transplantation, Vol. 21, Issue 4, pp. 657-69, 2013 (PubMed).

Kwiecinski, Elfimova, Noetel, Töx, Steffen, Hacker, Nischt, Dienes, Odenthal: "Expression of platelet-derived growth factor-C and insulin-like growth factor I in hepatic stellate cells is inhibited by miR-29." in: Laboratory investigation; a journal of technical methods and pathology, Vol. 92, Issue 7, pp. 978-87, 2012 (PubMed).

Zhong, Zhou, Ye, Cai, Zhang, Deng: "Hypoxia-inducible factor 1-?-AA-modified bone marrow stem cells protect PC12 cells from hypoxia-induced apoptosis, partially through VEGF/PI3K/Akt/FoxO1 pathway." in: Stem cells and development, Vol. 21, Issue 14, pp. 2703-17, 2012 (PubMed).

Kelly, Zhang, Wang, Zhang, Dominguez: "Intravenous renal cell transplantation for rats with acute and chronic renal failure." in: American journal of physiology. Renal physiology, Vol. 303, Issue 3, pp. F357-65, 2012 (PubMed).

Yilmaz, Ozaksit, Keskin, Tapisiz, Mollamahmutoglu, Uysal, Astarci, Ustun, Mulazımoglu: "The effect of formoterol on peritoneal VEGF levels in rats with endometriosis." in: Cytokine, Vol. 58, Issue 1, pp. 47-9, 2012 (PubMed).

Zaitone, Abo-Gresha: "Rosuvastatin promotes angiogenesis and reverses isoproterenol-induced acute myocardial infarction in rats: role of iNOS and VEGF." in: European journal of pharmacology, Vol. 691, Issue 1-3, pp. 134-42, 2012 (PubMed).

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