Rat VEGF PicoKine® Quick ELISA Kit

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.

Reproducibility To assay reproducibility, three samples with differing target protein concentrations were assayed using four different lots. Lots Lot 1 (pg/ml) Lot 2 (pg/ml) Lot 3 (pg/ml) Lot 4 (pg/ml) Mean (pg/ml) Standard Deviation CV (%) Sample 1 25 29 30 31 29 1.24 4.3% Sample 2 233 265 241 247 247 16.27 6.6% Sample 3 364 388 403 412 392 18.41 4.7% Lots Lot 1 (pg/ml) Lot 2 (pg/ml) Lot 3 (pg/ml) Lot 4 (pg/ml) Mean (pg/ml) Standard Deviation CV (%) Lots Lot 1 (pg/ml) Lot 2 (pg/ml) Lot 3 (pg/ml) Lot 4 (pg/ml) Mean (pg/ml) Standard Deviation CV (%) Sample 1 25 29 30 31 29 1.24 4.3% Sample 1 25 29 30 31 29 1.24 4.3% Sample 2 233 265 241 247 247 16.27 6.6% Sample 2 233 265 241 247 247 16.27 6.6% Sample 3 364 388 403 412 392 18.41 4.7% Sample 3 364 388 403 412 392 18.41 4.7%

Intra-Assay Precision Inter-Assay Precision Sample 1 2 3 1 2 3 n 16 16 16 24 24 24 Mean (pg/ml) 25 233 364 36 241 398 Standard deviation 1.4 18.87 22.93 2.45 19.04 21.89 CV (%) 5.6% 8.1% 6.3% 6.8% 7.9% 5.5% Intra-Assay Precision Inter-Assay Precision Sample 1 2 3 1 2 3 n 16 16 16 24 24 24 Mean (pg/ml) 25 233 364 36 241 398 Standard deviation 1.4 18.87 22.93 2.45 19.04 21.89 CV (%) 5.6% 8.1% 6.3% 6.8% 7.9% 5.5% Intra-Assay Precision Inter-Assay Precision Intra-Assay Precision Inter-Assay Precision Sample 1 2 3 1 2 3 Sample 1 2 3 1 2 3 n 16 16 16 24 24 24 n 16 16 16 24 24 24 Mean (pg/ml) 25 233 364 36 241 398 Mean (pg/ml) 25 233 364 36 241 398 Standard deviation 1.4 18.87 22.93 2.45 19.04 21.89 Standard deviation 1.4 18.87 22.93 2.45 19.04 21.89 CV (%) 5.6% 8.1% 6.3% 6.8% 7.9% 5.5% CV (%) 5.6% 8.1% 6.3% 6.8% 7.9% 5.5%

Description|Quantity|Volume Anti-tag Pre-coated 96-well Strip Microplate|1|12 strips of 8 wells Standard||2 Antibody A|1|3ml Antibody B|1|3ml Sample Diluent|1|15ml TBS-T Wash Buffer (25x) |1|12ml Color Developing Reagent (TMB) |1|10ml Stop Solution|1|10ml Adhesive Plate Sealers|2|Piece

Concentration (pg/ml) 0 15.6 31.2 62.5 125 250 500 1,000 O.D. 0.043 0.108 0.159 0.260 0.443 0.795 1.396 2.276 Concentration (pg/ml) 0 15.6 31.2 62.5 125 250 500 1,000 O.D. 0.043 0.108 0.159 0.260 0.443 0.795 1.396 2.276 Concentration (pg/ml) 0 15.6 31.2 62.5 125 250 500 1,000 Concentration (pg/ml) 0 15.6 31.2 62.5 125 250 500 1,000 O.D. 0.043 0.108 0.159 0.260 0.443 0.795 1.396 2.276 O.D. 0.043 0.108 0.159 0.260 0.443 0.795 1.396 2.276

Microplate Reader capable of reading absorbance at 450nm. Automated plate washer (optional) Pipettes and pipette tips capable of precisely dispensing 0.5 μl through 1 ml volumes of aqueous solutions. Multichannel pipettes are recommended for large amount of samples. Deionized or distilled water. 500ml graduated cylinders. Test tubes for dilution. Horizontal orbital microplate shaker capable of maintaining a speed of 500 rpm, amplitude 3mm.