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.
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Useful in Sandwich ELISA for Quantitative Detection of Antigen. Aliquot 0.1ml per well of the 10,000pg/ml, 5000pg/ml, 2500pg/ml, 1250pg/ml, 625pg/ml, 312pg/ml, 156pg/ml mouse Endostatin standard solutions into the precoated 96-well plate. Add 0.1ml of the sample diluent buffer into the control well (Zero well). Add 0.1ml of each properly diluted sample of mouse cell culture supernates, serum or plasma(heparin, EDTA) to each empty well. It is recommended that each mouse Endostatin standard solution and each sample be measured in duplicate.
Store vials at 4°C prior to opening. Centrifuge product if not completely clear after standing at room temperature. This product is stable for 6 months at 4°C as an undiluted liquid. Dilute only prior to immediate use. For extended storage freeze at -20°C or below for 12 months. Avoid cycles of freezing and thawing.