Rat Endostatin ELISA Kit PicoKine®

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 1 265 249 274 244 258 12.06 4.6% Sample 2 1669 1726 1859 1913 1791 98.25 5.4% Sample 3 4310 4545 4581 3831 4316 299.13 6.9% 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 1 265 249 274 244 258 12.06 4.6% Sample 1 265 249 274 244 258 12.06 4.6% Sample 2 1669 1726 1859 1913 1791 98.25 5.4% Sample 2 1669 1726 1859 1913 1791 98.25 5.4% Sample 3 4310 4545 4581 3831 4316 299.13 6.9% Sample 3 4310 4545 4581 3831 4316 299.13 6.9%

Intra-Assay Precision Inter-Assay Precision Sample 1 2 3 1 2 3 n 16 16 16 24 24 24 Mean (pg/ml) 265 1669 4310 253 1736 4538 Standard deviation 18.02 95.13 219.81 21.5 112.84 231.43 CV (%) 6.8% 5.7% 5.1%% 8.5% 6.5% 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) 265 1669 4310 253 1736 4538 Standard deviation 18.02 95.13 219.81 21.5 112.84 231.43 CV (%) 6.8% 5.7% 5.1%% 8.5% 6.5% 5.1% 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) 265 1669 4310 253 1736 4538 Mean (pg/ml) 265 1669 4310 253 1736 4538 Standard deviation 18.02 95.13 219.81 21.5 112.84 231.43 Standard deviation 18.02 95.13 219.81 21.5 112.84 231.43 CV (%) 6.8% 5.7% 5.1%% 8.5% 6.5% 5.1% CV (%) 6.8% 5.7% 5.1%% 8.5% 6.5% 5.1%

Description|Quantity Pre-coated 96-well strip microplate|1 Standard|2 vials Biotinylated antibody (100x) |100ul Avidin-Biotin-Peroxidase Complex (100x) |100ul Sample Diluent|30ml Antibody Diluent|12ml Avidin-Biotin-Peroxidase Diluent|12ml Color Developing Reagent (TMB) |10ml Stop Solution|10ml Wash Buffer (25x) |20ml Adhesive plate sealers|4

Store at 4°C for 6 months, at -20°C for 12 months. Avoid multiple freeze-thaw cycles (Ships with gel ice, can store for up to 3 days in room temperature. Freeze upon receiving.)

Concentration (pg/ml) 0 156 312 625 1250 2500 5000 10000 O.D. 0.018 0.099 0.162 0.296 0.522 0.853 1.462 2.093 Concentration (pg/ml) 0 156 312 625 1250 2500 5000 10000 O.D. 0.018 0.099 0.162 0.296 0.522 0.853 1.462 2.093 Concentration (pg/ml) 0 156 312 625 1250 2500 5000 10000 Concentration (pg/ml) 0 156 312 625 1250 2500 5000 10000 O.D. 0.018 0.099 0.162 0.296 0.522 0.853 1.462 2.093 O.D. 0.018 0.099 0.162 0.296 0.522 0.853 1.462 2.093

Microplate Reader capable of reading absorbance at 450nm. Incubator. 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.