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|Application / Reactivity||Mouse (Murine)|
|ELISA||21 ELISA Kits|
|Antigen||Vascular Endothelial Growth Factor A (VEGFA) ELISA Kits|
|Reactivity||Mouse (Murine) Alternatives|
Kits with alternative reactivity to:
|Method Type||Sandwich ELISA|
|Detection Range||2-1000 pg/mL|
|Minimum Detection Limit||2 pg/mL|
|20 references available|
|Supplier||Log in to see|
Product Details VEGF ELISA KitTarget details Application Details Handling References for VEGF Kit (ABIN625186) Images
|Purpose||Mouse VEGF-A ELISA Kit for cell culture supernatants, plasma, and serum samples.|
|Sample Type||Plasma, Cell Culture Supernatant, Serum|
|Specificity||This ELISA kit shows no cross-reactivity with any of the cytokines tested: Mouse 6Ckine, CTACK, Eotaxin, GCSF, GM-CSF, IL-2, IL-3, IL-4, IL-5, IL-6, IL-9, IL-10, IL-12p40, IL-12p70, IL-13, IL-17, IFN-gamma, KC, Leptin, MCP-5, MIP-1 alpha, MIP-2, MIP-3 beta, RANTES, SCF, sTNFri, TARC, TIMP-1, TNF-alpha, Tpo.|
|Cross-Reactivity (Details)||This ELISA kit shows no cross-reactivity with any of the cytokines tested (e.g., Mouse 6Ckine, CTACK, Eotaxin, GCSF, GM-CSF, IL-2, IL-3, IL-4, IL-5, IL-6, IL-9, IL-10, IL-12p40, IL-12p70, IL-13, IL-17, IFN-gamma, KC, Leptin, MCP-5, MIP-1alpha, MIP-2, MIP-3beta, RANTES, SCF, sTNFri, TARC, TIMP-1, TNF-alpha, Tpo).|
|Sensitivity||< 2 pg/mL|
|Material not included||
Target detailsProduct Details VEGF ELISA Kit Application Details Handling References for VEGF Kit (ABIN625186) Images back to top
|Alternative Name||VEGF-A (VEGFA ELISA Kit Abstract)|
|Background||Vascular endothelial growth factor A (VEGF-A) (Vascular permeability factor) (VPF)|
|Research Area||Cytokines, Growth Factors, Receptors, Immunology, Inflammation, Signaling|
|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|
Application DetailsProduct Details VEGF ELISA Kit Target details Handling References for VEGF Kit (ABIN625186) Images back to top
|Application Notes||Recommended Dilution for serum and plasma samples3 fold|
|Sample Volume||100 μL|
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: 3 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 should be diluted 5-fold with deionized or distilled water.
4. Preparation of standard: Briefly spin the vial of Item C. 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 25 ng/mL standard. Dissolve the powder thoroughly by a gentle mix. Add 40 µL VEGF standard from the vial of Item C, into a tube with 960 µL Assay Diluent A or 1x Assay Diluent B to prepare a 1,000 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 200 µL 200 µL 200 µL 200 µL 40 µL standard + 960.0 µL 200myl 1,000 400 160 64 25.6 10.2 4.1 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 160-fold with 1x Assay Diluent B. For example: Briefly spin the vial (Item G) and pipette up and down to mix gently . Add 50 µL of HRP-Streptavidin concentrate into a tube with 8 ml 1x Assay Diluent B to prepare a 160-fold diluted HRP-Streptavidin solution (don't store the diluted solution for next day use). Mix well.
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 Mouse VEGF concentration (pg/mL) 1 10 100 1000 10000 O D =4 50 n m 0.01 0.1 1 10 Assay Diluent B Mouse VEGF concentration (pg/mL) 1 10 100 1000 10000 O D =4 50 n m 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 mouse VEGF into mouse serum, plasma and cell culture media. Mean recoveries are as follows: Sample Type Average % Recovery Range ( %) Serum 87.67 79-102 Plasma 89.32 81-103 Cell culture media 94.29 83-105
Linearity: Sample Type Serum Plasma Cell Culture Media 1:2 Average % of Expected 96 97 95 Range ( %) 83-103 84-103 82-102 1:4 Average % of Expected 94 98 97 Range ( %) 82-102 84-104 83-103
Reproducibility: Intra-Assay: CV<10 % Inter-Assay: CV<12 %
|Assay Precision||Intra-Assay: CV< 10 % Inter-Assay: CV< 12 %|
|Restrictions||For Research Use only|
HandlingProduct Details VEGF ELISA Kit Target details Application Details References for VEGF Kit (ABIN625186) Images back to top
|Handling Advice||Avoid repeated freeze-thaw cycles.|
|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|
References for VEGF Kit (ABIN625186)Product Details VEGF ELISA Kit Target details Application Details Handling Images back to top
|Product cited in:||
Huey, Smith, Sulaeman, Breen: "Skeletal myofiber VEGF is necessary for myogenic and contractile adaptations to functional overload of the plantaris in adult mice." in: Journal of applied physiology (Bethesda, Md. : 1985), Vol. 120, Issue 2, pp. 188-95, 2016
Li, Wang, Huang, Zhang, Yao, Wang, Lv, An, Corrigan, Huang, Ying: "Distinct sustained structural and functional effects of interleukin-33 and interleukin-25 on the airways in a murine asthma surrogate." in: Immunology, Vol. 145, Issue 4, pp. 508-18, 2015
Pala, Atilgan, Ozkan, Kavak, Ilhan, Akpolat, Sapmaz: "Effect of varying doses of tamoxifen on ovarian histopathology, serum VEGF, and endothelin 1 levels in ovarian hyperstimulation syndrome: an experimental study." in: Drug design, development and therapy, Vol. 9, pp. 1761-6, 2015
Patel, Thaker: "The effects of A2B receptor modulators on vascular endothelial growth factor and nitric oxide axis in chronic cyclosporine nephropathy." in: Journal of pharmacology & pharmacotherapeutics, Vol. 6, Issue 3, pp. 147-53, 2015
Martin, Nguyen, Molinolo, Gutkind: "Accumulation of dephosphorylated 4EBP after mTOR inhibition with rapamycin is sufficient to disrupt paracrine transformation by the KSHV vGPCR oncogene." in: Oncogene, Vol. 33, Issue 18, pp. 2405-12, 2014
Wei, Yu, Lee, Chen, Taylor, Deveau, Yu, Wei: "Regulatory role of the JNK-STAT1/3 signaling in neuronal differentiation of cultured mouse embryonic stem cells." in: Cellular and molecular neurobiology, Vol. 34, Issue 6, pp. 881-93, 2014
Abbas, Borman, Terzi, Terzi, Bayraktar, Ozkan, Yazc: "Inhibition of the Notch Pathway Promotes Flap Survival by Inducing Functional Neoangiogenesis." in: Annals of plastic surgery, 2014
Cai, Zhou, Zhou, Lou: "Hypoxic conditioned medium from rat cerebral cortical cells enhances the proliferation and differentiation of neural stem cells mainly through PI3-K/Akt pathways." in: PLoS ONE, Vol. 9, Issue 11, pp. e111938, 2014
Yamamoto, Fujita, Tanaka, Kojima, Kanatani, Ishihara, Tachibana: "Low oxygen tension enhances proliferation and maintains stemness of adipose tissue-derived stromal cells." in: BioResearch open access, Vol. 2, Issue 3, pp. 199-205, 2013
Hartwich, Orr, Ng, Spence, Morton, Davidoff: "HIF-1? activation mediates resistance to anti-angiogenic therapy in neuroblastoma xenografts." in: Journal of pediatric surgery, Vol. 48, Issue 1, pp. 39-46, 2013
Said, Frierson, Sanchez-Carbayo, Brekken, Theodorescu: "Loss of SPARC in bladder cancer enhances carcinogenesis and progression." in: The Journal of clinical investigation, Vol. 123, Issue 2, pp. 751-66, 2013
Saldaña, Crespo, Bensiamar, Arruebo, Vilaboa: "Mechanical forces regulate stem cell response to surface topography." in: Journal of biomedical materials research. Part A, 2013
Xiao, Guo, Chen, El-Gohary, Wiersch, Gaffar, Prasadan, Shiota, Gittes: "Hypoglycemia reduces vascular endothelial growth factor A production by pancreatic beta cells as a regulator of beta cell mass." in: The Journal of biological chemistry, Vol. 288, Issue 12, pp. 8636-46, 2013
Brunner, Huber, Weinberger, Vallaster, Wollenweber, Gerbitz, Hacker, Franz: "Migration of bone marrow-derived cells and improved perfusion after treatment with erythropoietin in a murine model of myocardial infarction." in: Journal of cellular and molecular medicine, Vol. 16, Issue 1, pp. 152-9, 2012
Juffer, Jaspers, Lips, Bakker, Klein-Nulend: "Expression of muscle anabolic and metabolic factors in mechanically loaded MLO-Y4 osteocytes." in: American journal of physiology. Endocrinology and metabolism, Vol. 302, Issue 4, pp. E389-95, 2012
George, Rajoria, Suriano, Mittleman, Tiwari: "Hypoxia and estrogen are functionally equivalent in breast cancer-endothelial cell interdependence." in: Molecular cancer, Vol. 11, pp. 80, 2012
Altman, Nguyen, Patel, Fambrough, Beedle, Hardman, Murph: "Regulator of G-Protein Signaling 5 Reduces HeyA8 Ovarian Cancer Cell Proliferation and Extends Survival in a Murine Tumor Model." in: Biochemistry research international, Vol. 2012, pp. 518437, 2012
Jones, Antonelli, Masko, Broadwater, Lascola, Fels, Dewhirst, Dyck, Nagendran, Flores, Betof, Nelson, Pollak, Dash, Young, Freedland: "Exercise modulation of the host-tumor interaction in an orthotopic model of murine prostate cancer." in: Journal of applied physiology (Bethesda, Md. : 1985), Vol. 113, Issue 2, pp. 263-72, 2012
Sawai, Murata, Koto, Matsui, Horie, Ashihara, Maekawa, Fushiki, Kubo: "Effects of low-intensity pulsed ultrasound on osteosarcoma and cancer cells." in: Oncology reports, Vol. 28, Issue 2, pp. 481-6, 2012
Clarke, Al Ahmad, Lee, Parham, Auckland, Fertala, Kahle, Shaw, Roberts, Bix: "Perlecan Domain V induces VEGf secretion in brain endothelial cells through integrin α5β1 and ERK-dependent signaling pathways." in: PLoS ONE, Vol. 7, Issue 9, pp. e45257, 2012
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