Mouse Endostatin PicoKine® Quick ELISA Kit

Endostatin is a naturally-occurring 20-kDa C-terminal fragment derived from type XVIII collagen. It is reported to serve as an anti-angiogenic agent, similar to angiostatin and thrombospondin. And It is produced by proteolytic cleavage of collagen XVIII, a member of the multiplexin family that is characterized by interruptions in the triple helix creating multiple domains, by proteases such as cathepsins. Using a genomic clone as a probe for fluorescence in situ hybridization, Endostatin was mapped the COL18A1 gene to 21q22.3. By immunoprecipitation analysis using membrane fractions of human mammary epithelial cells, It showed that endostatin specifically bound to cell surface nucleolin with high affinity. Blockage of nucleolin with neutralizing antibody or knockdown of nucleolin by RNA interference countered the antiendothelial activity of endostatin and abrogated its antiangiogenic and antitumor activity in vivo.

Lots Lot 1 (pg/ml) Lot 2 (pg/ml) Lot 3 (pg/ml) Lot 4 (pg/ml) Mean (pg/ml) Standard Deviation CV (%) Sample 1 305 306 299 312 306 13.44 4.4% Sample 2 1523 1512 1399 1539 1493 68.69 4.6% Sample 3 4766 4723 3964 4369 4456 227.23 5.1% 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 1 305 306 299 312 306 13.44 4.4% Sample 1 305 306 299 312 306 13.44 4.4% Sample 2 1523 1512 1399 1539 1493 68.69 4.6% Sample 2 1523 1512 1399 1539 1493 68.69 4.6% Sample 3 4766 4723 3964 4369 4456 227.23 5.1% Sample 3 4766 4723 3964 4369 4456 227.23 5.1%

Intra-Assay Precision Inter-Assay Precision Sample 1 2 3 1 2 3 n 16 16 16 24 24 24 Mean (pg/ml) 305 1523 4766 304 1462 4785 Standard deviation 12.2 74.63 314.56 13.07 74.56 282.32 CV (%) 4% 4.9% 6.6% 4.3% 5.1% 5.9% Intra-Assay Precision Inter-Assay Precision Sample 1 2 3 1 2 3 n 16 16 16 24 24 24 Mean (pg/ml) 305 1523 4766 304 1462 4785 Standard deviation 12.2 74.63 314.56 13.07 74.56 282.32 CV (%) 4% 4.9% 6.6% 4.3% 5.1% 5.9% 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) 305 1523 4766 304 1462 4785 Mean (pg/ml) 305 1523 4766 304 1462 4785 Standard deviation 12.2 74.63 314.56 13.07 74.56 282.32 Standard deviation 12.2 74.63 314.56 13.07 74.56 282.32 CV (%) 4% 4.9% 6.6% 4.3% 5.1% 5.9% CV (%) 4% 4.9% 6.6% 4.3% 5.1% 5.9%

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) 0 156 312 625 1,250 2,500 5,000 10,000 O.D. 0.002 0.109 0.186 0.368 0.747 1.290 1.917 2.267 Concentration (pg/ml) 0 156 312 625 1,250 2,500 5,000 10,000 O.D. 0.002 0.109 0.186 0.368 0.747 1.290 1.917 2.267 Concentration (pg/ml) 0 156 312 625 1,250 2,500 5,000 10,000 Concentration (pg/ml) 0 156 312 625 1,250 2,500 5,000 10,000 O.D. 0.002 0.109 0.186 0.368 0.747 1.290 1.917 2.267 O.D. 0.002 0.109 0.186 0.368 0.747 1.290 1.917 2.267

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.