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Catalog No: OKBB00272
Size:96 Tests
Price: $583.00
SKU
OKBB00272
Availability: Domestic: within 1-2 week delivery | International: within 1-2 week delivery
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Datasheets/ManualsClick here to download product manual. As variation between lots may occur, always reference the lot-specific manual received with each kit.
Product Info
Predicted Species ReactivityRat
ApplicationELISA
ELISA Kit Detection MethodColorimetric, OD450 nm
ELISA Kit Duration~ 3 Hours
ELISA Kit PrincipleAviva's rat VEGF ELISA Kit was based on standard sandwich enzyme-linked immune-sorbent assay technology. A monoclonal antibody from mouse specific for VEGF has been precoated onto 96-well plates. Standards (NS0,A27-R190) and test samples are added to the wells, a biotinylated detection polyclonal antibody from goat specific for VEGF is added subsequently and then followed by washing with PBS or TBS buffer. Avidin-Biotin-Peroxidase Complex was added and unbound conjugates were washed away with PBS or TBS buffer. HRP substrate TMB was used to visualize HRP enzymatic reaction. TMB was catalyzed by HRP to produce a blue color product that changed into yellow after adding acidic stop solution. The absorbance of yellow is proportional to the rat VEGF amount of sample captured in plate.
ELISA Kit Range15.6 pg/ml - 1,000 pg/ml
ELISA Kit Reproducibility
Intra-Assay PrecisionInter-Assay Precision
Sample123123
Mean (pg/mL)120305621236289632
St. Dev.4.4415.8629.8110.6218.2134.76
%CV3.75.24.804.56.305.5
ELISA Kit Component
ComponentAmount
Lyophilized recombinant rat VEGF standard10 ng/tube x 2
Anti-rat VEGF Antibody Well Plate96 Wells
Sample diluent buffer30 mL
Biotinylated anti-rat VEGF antibody130 uL, dilution 1:100
Antibody diluent buffer12 mL
Avidin-Biotin-Peroxidase Complex (ABC)130 uL, dilution 1:100
ABC diluent buffer12 mL
TMB color developing agent10 mL
TMB stop solution10 mL
Additional InformationRange: 15.6pg/ml-1000pg/ml
::Principle: Aviva’s rat VEGF ELISA Kit was based on standard sandwich enzyme-linked immune-sorbent assay technology. Rat VEGF specific-specific monoclonal antibodies were precoated onto 96-well plates. The rat specific detection polyclonal antibodies were biotinylated. The test samples and biotinylated detection antibodies were added to the wells subsequently and then followed by washing with PBS or TBS buffer. Avidin-Biotin-Peroxidase Complex was added and unbound conjugates were washed away with PBS or TBS buffer. HRP substrate TMB was used to visualize HRP enzymatic reaction. TMB was catalyzed by HRP to produce a blue color product that changed into yellow after adding acidic stop solution. The density of yellow is proportional to the rat VEGF amount of sample captured in plate.
::Notes: 1. To inspect the validity of experiment operation and the appropriateness of sample dilution proportion, pilot
experiment using standards and a small number of samples is recommended.
2. The TMB Color Developing agent is colorless and transparent before using, contact us freely if it is not the case.
3. Before using the Kit, spin tubes and bring down all components to the bottom of tubes.
4. Duplicate well assay is recommended for both standard and sample testing.
5. Don’t let 96-well plate dry, for dry plate will inactivate active components on plate.
6. Don’t reuse tips and tubes to avoid cross contamination.
7. To avoid to use the reagents from different batches together.
8. In order to avoid marginal effect of plate incubation due to temperature difference ( reaction may be stronger in the
marginal wells), it is suggested that the diluted ABC and TMB solution will be pre-warmed in 37C for 30 min before
using.
Background: Vascular permeability factor/vascular endothelial growth factor (VPF/VEGF), a potent cytokine expressed by most malignant tumors, has critical roles in vasculogenesis and both physiological and pathological angiogenesis.VEGF produced by tumor cells potently stimulates endothelial cell proliferation and angiogenesis and plays a key role in the pathophysiology of several neoplasias.VEGF may also play a pivotal role in mediating the development and progression of diabetic retinopathy. VEGF, a major regulator of angiogenesis, binds to two receptor tyrosine kinases, KDR/Flk-1 and Flt-1. The VEGF gene is mapped by fluorescence in situ hybridization to chromosome 6p12.The standard product used in this kit is recombinant rat VEGF164, which is a 25KDa single chain as well as a 50KDa dimer.
Reconstitution and StorageStore at 4C for 6 months, at -20C for 12 months. Avoid multiple freeze-thaw cycles (Shipped with wet ice.)
Sample Typecell culture supernates, serum and plasma (heparin, EDTA, citrate)
Sensitivity<1 pg/ml
Predicted Homology Based on Immunogen SequenceNo detectable cross-reactivity with any other cytokine.
Reference1. Basu, S.; Nagy, J. A.; Pal, S.; Vasile, E.; Eckelhoefer, I. A.; Bliss, V. S.; Manseau, E. J.; Dasgupta, P. S.; Dvorak, H.
F.; Mukhopadhyay, D. The neurotransmitter dopamine inhibits angiogenesis induced by vascular permeability
factor/vascular endothelial growth factor. Nature Med. 7: 569-574, 2001.
2. Poulaki, V.; Mitsiades, C. S.; McMullan, C.; Sykoutri, D.; Fanourakis, G.; Kotoula, V.; Tseleni-Balafouta, S.; Koutras,
D. A.; Mitsiades, N. Regulation of vascular endothelial growth factor expression by insulin-like growth factor I in
thyroid carcinomas. J. Clin. Endocr. Metab. 88: 5392-5398, 2003.
3. Awata, T.; Inoue, K.; Kurihara, S.; Ohkubo, T.; Watanabe, M.; Inukai, K.; Inoue, I.; Katayama, S. A common
polymorphism in the 5-prime-untranslated region of the VEGF gene is associated with diabetic retinopathy in type 2
diabetes. Diabetes 51: 1635-1639, 2002.
4. Soker, S.; Takashima, S.; Miao, H. Q.; Neufeld, G.; Klagsbrun, M. Neuropilin-1 is expressed by endothelial and
tumor cells as an isoform-specific receptor for vascular endothelial growth factor. Cell 92: 735-745, 1998.
5. Wei, M.-H.; Popescu, N. C.; Lerman, M. I.; Merrill, M. J.; Zimonjic, D. B. Localization of the human vascular
endothelial growth factor gene, VEGF, at chromosome 6p12. Hum. Genet. 97: 794-797, 1996.
SpecificityNatural and recombinant rat VEGF
ImmunogenExpression system for standard: NS0; Immunogen sequence: A27-R190
DescriptionFor quantitative detection of rat VEGF in cell culture supernatants, serum and plasma(heparin, EDTA, citrate).
Gene SymbolVEGFA
Gene Full Namevascular endothelial growth factor A
Alias SymbolsMGC70609, MVCD1, Vascular endothelial growth factor, Vascular endothelial growth factor A, Vascular permeability factor, VEGF A, VEGFA, VPF
NCBI Gene Id83785
Protein NameVascular endothelial growth factor A
Description of TargetVascular permeability factor/vascular endothelial growth factor (VPF/VEGF), a potent cytokine expressed by most malignant tumors, has critical roles in vasculogenesis and both physiological and pathological angiogenesis.VEGF produced by tumor cells potently stimulates endothelial cell proliferation and angiogenesis and plays a key role in the pathophysiology of several neoplasias.VEGF may also play a pivotal role in mediating the development and progression of diabetic retinopathy. VEGF, a major regulator of angiogenesis, binds to two receptor tyrosine kinases, KDR/Flk-1 and Flt-1. The VEGF gene is mapped by fluorescence in situ hybridization to chromosome 6p12.The standard product used in this kit is recombinant rat VEGF164, which is a 25KDa single chain as well as a 50KDa dimer.
Uniprot IDP16612
Protein Accession #NP_001103804.1
Nucleotide Accession #NM_001110334.2
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