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Canine VEGF ELISA Kit (CNFI00022)

SKU:
CNFI00022
Product Type:
ELISA Kit
Size:
96 Assays
Uniprot:
Q9MYV3
Sensitivity:
18.75pg/ml
Range:
31.25-2000pg/ml
ELISA Type:
Sandwich
Synonyms:
VEGF, VAS, Vasculotropin, VEGFA, VPF, MVCD1, vascular endothelial growth factor A, Vascular permeability factor, VEGF-A, VPFvascular endothelial growth factor
Reactivity:
Canine
€649
Frequently bought together:

Description

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Canine VEGF ELISA Kit

The Canine VEGF (Vascular Endothelial Growth Factor) ELISA Kit is a valuable tool for the precise measurement of VEGF levels in canine samples including serum, plasma, and tissue culture supernatants. This kit offers superior sensitivity and specificity, ensuring accurate and consistent results for a variety of research applications.VEGF is a key signaling protein involved in the formation of new blood vessels, a process known as angiogenesis. It plays a critical role in various physiological processes, including wound healing, organ development, and tumor growth.

By measuring VEGF levels, researchers can gain insights into the angiogenic activity in canine tissues and the role it plays in different diseases and conditions.This canine VEGF ELISA kit provides researchers with a reliable and efficient tool for studying angiogenesis in dogs, allowing for a better understanding of the mechanisms underlying vascular development and the potential implications for canine health and disease.

Product Name:Canine VEGF (Vascular Endothelial cell Growth Factor) ELISA Kit
Product Code:CNFI00022
Size:96 Assays
Target:Canine VEGF
Alias:VEGF, VAS, Vasculotropin, VEGFA, VPF, MVCD1, vascular endothelial growth factor A, Vascular permeability factor, VEGF-A, VPFvascular endothelial growth factor
Reactivity:Canine
Detection Method:Sandwich ELISA, Double Antibody
Sensitivity:18.75pg/ml
Range:31.25-2000pg/ml
Storage:4°C for 6 months
Note:For Research Use Only
Recovery:Matrices listed below were spiked with certain level of Canine VEGF and the recovery rates were calculated by comparing the measured value to the expected amount of Canine VEGF in samples.
MatrixRecovery range(%)Average(%)
serum(n=5)85-10294
EDTA plasma(n=5)91-10399
UFH plasma(n=5)85-10292
Linearity:The linearity of the kit was assayed by testing samples spiked with appropriate concentration of Canine VEGF and their serial dilutions. The results were demonstrated by the percentage of calculated concentration to the expected.
Sample1:21:41:8
serum(n=5)85-102%90-100%97-103%
EDTA plasma(n=5)86-96%83-99%82-100%
UFH plasma(n=5)87-98%81-96%92-100%
Intra-Assay:CV <8%
Inter-Assay:CV <10%
Uniprot:Q9MYV3
UniProt Protein Function:Growth factor active in angiogenesis, vasculogenesis and endothelial cell growth. Induces endothelial cell proliferation, promotes cell migration, inhibits apoptosis and induces permeabilization of blood vessels. Binds to the FLT1/VEGFR1 and KDR/VEGFR2 receptors, heparan sulfate and heparin. Binding to NRP1 receptor initiates a signaling pathway needed for motor neuron axon guidance and cell body migration, including for the caudal migration of facial motor neurons from rhombomere 4 to rhombomere 6 during embryonic development.
UniProt Code:Q9MYV3
NCBI GenInfo Identifier:160358860
NCBI Gene ID:403802
NCBI Accession:NP_001003175.2
UniProt Secondary Accession:Q9MYV3,Q9XSF3, Q9XSF4, Q9XSF5,
UniProt Related Accession:Q9MYV3
Molecular Weight:22,292 Da
NCBI Full Name:vascular endothelial growth factor A isoform 1
NCBI Synonym Full Names:vascular endothelial growth factor A
NCBI Official Symbol:VEGFA  
NCBI Official Synonym Symbols:VEGF  
NCBI Protein Information:vascular endothelial growth factor A
UniProt Protein Name:Vascular endothelial growth factor A
UniProt Synonym Protein Names:Vascular permeability factor; VPF
UniProt Gene Name:VEGFA  
StepProcedure
1. Set standard, test sample and control (zero) wells on the pre-coated plate respectively, and then, record their positions. It is recommended to measure each standard and sample in duplicate. Wash plate 2 times before adding standard, sample and control (zero) wells!
2.Aliquot 0.1ml standard solutions into the standard wells.
3. Add 0.1 ml of Sample / Standard dilution buffer into the control (zero) well.
4. Add 0.1 ml of properly diluted sample ( Human serum, plasma, tissue homogenates and other biological fluids.) into test sample wells.
5.Seal the plate with a cover and incubate at 37°C for 90 min.
6. Remove the cover and discard the plate content, clap the plate on the absorbent filter papers or other absorbent material. Do NOT let the wells completely dry at any time. Wash plate X2.
7. Add 0.1 ml of Biotin- detection antibody working solution into the above wells (standard, test sample & zero wells). Add the solution at the bottom of each well without touching the side wall.
8.Seal the plate with a cover and incubate at 37°C for 60 min.
9. Remove the cover, and wash plate 3 times with Wash buffer. Let wash buffer rest in wells for 1 min between each wash.
10. Add 0.1 ml of SABC working solution into each well, cover the plate and incubate at 37°C for 30 min.
11. Remove the cover and wash plate 5 times with Wash buffer, and each time let the wash buffer stay in the wells for 1-2 min.
12. Add 90 µL of TMB substrate into each well, cover the plate and incubate at 37°C in dark within 10-20 min. (Note: This incubation time is for reference use only, the optimal time should be determined by end user.) And the shades of blue can be seen in the first 3-4 wells (with most concentrated standard solutions), the other wells show no obvious color.
13. Add 50 µL of Stop solution into each well and mix thoroughly. The color changes into yellow immediately.
14. Read the O.D. absorbance at 450 nm in a microplate reader immediately after adding the stop solution.

When carrying out an ELISA assay it is important to prepare your samples in order to achieve the best possible results. Below we have a list of procedures for the preparation of samples for different sample types.

Sample Type Protocol
Serum:

If using serum separator tubes, allow samples to clot for 30 minutes at room temperature. Centrifuge for 10 minutes at 1,000x g. Collect the serum fraction and assay promptly or aliquot and store the samples at -80°C. Avoid multiple freeze-thaw cycles.

If serum separator tubes are not being used, allow samples to clotovernight at 2-8°C. Centrifuge for 10 minutes at 1,000x g. Removeserum and assay promptly or aliquot and store the samples at-80°C. Avoid multiple freeze-thaw cycles.

Plasma: Collect plasma using EDTA or heparin as an anti-coagulant. Centrifuge samples at 4°C for 15 mins at 1000 × g within 30 mins of collection. Collect the plasma fraction and assay promptly or aliquot and store the samples at -80°C. Avoid multiple freeze-thaw cycles.Note: Over haemolysed samples are not suitable for use with this kit.
Urine & Cerebrospinal Fluid: Collect the urine (mid-stream) in a sterile container, centrifuge for 20 mins at 2000-3000 rpm. Remove supernatant and assay immediately. If any precipitation is detected, repeat the centrifugation step. A similar protocol can be used for cerebrospinal fluid.
Cell Culture Supernatant: Collect the cell culture media by pipette, followed by centrifugation at 4°C for 20 mins at 1500 rpm. Collect the clear supernatant and assay immediately.
Cell Lysates: Solubilize cells in lysis buffer and allow to sit on ice for 30 minutes. Centrifuge tubes at 14,000 x g for 5 minutes to remove insoluble material. Aliquot the supernatant into a new tube and discard the remaining whole cell extract. Quantify total protein concentration using a total protein assay. Assay immediately or aliquot and store at ≤ -20°C.
Tissue Homogenates: The preparation of tissue homogenates will vary depending upon tissue type. Rinse tissue with 1X PBS to remove excess blood & homogenizein 20ml of 1X PBS (including protease inhibitors) and store overnight at ≤ -20°C. Two freeze-thaw cycles are required to break the cell membranes. To further disrupt the cell membranes you can sonicate the samples. Centrifuge homogenates for 5 mins at 5000xg. Remove the supernatant and assay immediately or aliquot and store at -20°C or-80°C.
Tissue Lysates: Rinse tissue with PBS, cut into 1-2 mm pieces, and homogenize with a tissue homogenizer in PBS. Add an equal volume of RIPA buffer containing protease inhibitors and lyse tissues at room temperature for 30 minutes with gentle agitation. Centrifuge to remove debris. Quantify total protein concentration using a total protein assay. Assay immediately or aliquot and store at ≤ -20 °C.
Breast Milk: Collect milk samples and centrifuge at 10,000 x g for 60 min at 4°C. Aliquot the supernatant and assay. For long term use, store samples at -80°C. Minimize freeze/thaw cycles.

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