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LPL ELISA
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■Application Quantification of LPL in human plasma ■Assay procedure for MARKIT-M LPL
■Contents of MARKIT-M LPL ・Standard 0 (lyophilized): 1 vial (for 0.5 mL). ・Standard 25 (lyophilized): 1 vial (for 0.5 mL) contains: T-LPL* 12.5 ng. ・Standard 50 (lyophilized): 1 vial (for 0.5 mL) contains: T-LPL* 25 ng. ・Standard100 (lyophilized): 1 vial (for 0.5 mL) contains: T-LPL* 50 ng. ・Standard 200 (lyophilized): 1 vial (for 0.5 mL) contains: T-LPL* 100 ng. ・Standard 300 (lyophilized): 1 vial (for 0.5 mL) contains: T-LPL* 150 ng. ・Stabilizer solution (bottle No. 1): 1 bottle (14 mL). ・Wash buffer concentrate (bottle No. 2): 1 bottle (90 mL). ・LPL antibody-enzyme conjugate (bottle No. 3): 1 bottle (14 mL). Each mL contains: HRP-labeled anti-human LPL monoclonal antibody. ・LPL antibody-coated wells: 1 plate (96 wells). Each well contains: anti-human LPL monoclonal antibody. ・Substrate tablet: 3. One tablet contains: o-phenylenediamine dihydrochloride (OPD) (13 mg). ・Substrate diluent (bottle No. 4): 3 bottles (15 mL each). One bottle contains: hydrogen peroxide (15μL). ・Stop solution (bottle No. 5): 1 bottle (15 mL). ・Microplate for dilution: 1 plate (96 wells). ・Graph paper: 1 sheet.
■Judgment
of the result of determination
1. Reproducibility When two distinct samples (LPL, 100-240 ng/mL) are determined 10 times each simultaneously, the coefficient of variation in their absorbances should be less than 5%. 2. Assay range LPL 3.6 -300 ng/mL ■Correlation between MARKIT-M LPL and LPL enzyme activity The correlation factor (r) between the LPL mass (X) measured by MARKIT-M LPL and the LPL activity (Y) determined by selective immunoinactivation assay was 0.945, and the regression was Y=0.05X-0.36 (n=33). ■Storage method and expiry period Storage: Store in a cool place (2-10oC), protected from light. Avoid freezing. Expiry period: 2 years ■References 1) Ikeda, Y., et al.: Biochim.Biophys.Acta 1003: 254, 1989. 2) Ikeda, Y., et al.: J. Lipid Res. 31: 1911, 1990. 3) Kimura, H., et al.: Clin. Biochem. 32: 15, 1999. 4) Takagi, A., et al.: J. Clin. Invest. 89: 581, 1992. 5) Takagi, A., et al.: J. Lipid Res. 35: 2008, 1994. 6) Antikainen, M, et al.: Eur. J. Clin. Clin. Biochem. 34: 547, 1996. 7) Suga, S., et al.: J. Intern. Med. 243: 317, 1998. 8) Takagi, A., et al.: Clin. Chim. Acta 285: 143, 1999. 9) Takagi, A., et al.: Biochim. Biophys. Acta 1502: 433, 2000. 10) Ikeda, Y., et al.: Clin. Sci. 99: 569, 2000. 11) Hoffmann, M.M, et al.: J. Clin. Endocr. Metab. 85: 4795, 2000. 12) Ikeda, Y., et al.: J. Lipid Res. 42: 1072, 2001. 13) Ikeda, Y., et al.: Clin. Chim. Acta 316: 179, 2002. 14) Gotto, A.M.: Am. J. Cardiol. 82: 22Q, 1998. 15) Ikeda, Y, et al.: 2nd International Symposium on Triglycerides and HDL: Role in cardiovascular disease and the metabolic syndrome (New York, USA), 2005. | ||||||||||||||||||||||||||||
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