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73871-2 Markit M-LPL 96wells

For research use only

Lipoprotein lipase (LPL) is

a key enzyme for the

metabolism of triglyceride (TG)-rich lipoproteins, and functionally

it hydrolyzes the TGs in chylomicrons and very low-density lipoprotein (VLDL) in

the blood.  It is well known that continual hyperlipoproteinemia is a high risk factor for arteriosclerosis and myocardial infarction.  In order to investigate functional abnormalities of LPL in genetic disorders and in secondary hyperlipoproteinemia, measurement of LPL is very important, and it is also very informative for medical treatment of hyperlipoproteinemia. MARKIT-M LPL is a direct sandwich enzyme-linked immunosorbent assay (ELISA) for the quantification of LPL in human plasma using two distinct anti-human LPL monoclonal antibodies.

 

■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

Reference data of LPL concentration in preheparin plasma (PreHP) and PHP using 30 IU/kg of heparin

 

 

 

■Performance

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.