SCETI established in 1952, develops, distributes and manufactures worldwide Assay kits for clinical and research applications. Obesity/Diabetes; Adiponectin, Ghrelin, Glucagon, GLP-1, LPL, etc, Rheumatism; MMP3, Pneumonia; KL6, Inflammation; PTX3, Infection; Flu, Pancreas cancer marker; coming soon, various characteristic antibodies, and other significant products in the wide-ranging field.　 Please stop by our booth at “Booth No. 1207”. We are looking forward to meeting you soon. Thank you.

July 27-29,2010

Anaheim Convention Center (Anaheim/CA,USA)

Booth No.

1207

Introduction

For study with rat and mouse, glucagon assay with smaller volume of specimens are comfortable. YK090 Glucagon EIA can use with not only 100uL specimens but also 50uL specimens. So we compared performance of YK090 Glucagon EIA with 100uL and 50uL specimens.

Method

Glucagon assay performance between with 100uL specimens and 50uL specimens are examined. Each assay are placed along with package insert (Ⅳ. Method).

Result

Standard curve(A490)

Glucagon value(pg/mL)

Discussion

Assay results with 50uL specimens are nearly same as assay results with 100uL specimens (standard procedure). But specimen volume should be fixed in a sequential study.

Here is the publication information;

Endocrinology. 2009 Nov;150(11):5113-8. Epub 2009 Oct 9.
“The RAPID method for blood processing yields new insight in plasma concentrations and molecular forms of circulating gut peptides.”
Stengel A, Keire D, Goebel M, Evilevitch L, Wiggins B, Taché Y, Reeve JR Jr.

Abstract:

The correct identification of circulating molecular forms and measurement of peptide levels in blood entails that the endocrine peptide being studied is stable and recovered in good yields during blood processing. However, it is not clear whether this is achieved in studies using standard blood processing. Therefore, we compared peptide concentration and form of 12 (125)I-labeled peptides using the standard procedure (EDTA-blood on ice) and a new method employing Reduced temperatures, Acidification, Protease inhibition, Isotopic exogenous controls, and Dilution (RAPID). During standard processing there was at least 80% loss for calcitonin-gene-related peptide and cholecystokinin-58 (CCK-58) and more than 35% loss for amylin, insulin, peptide YY forms (PYY((1-36)) and PYY((3-36))), and somatostatin-28. In contrast, the RAPID method significantly improved the recovery for 11 of 12 peptides (P < 0.05) and eliminated the breakdown of endocrine peptides occurring after standard processing as reflected in radically changed molecular forms for CCK-58, gastrin-releasing peptide, somatostatin-28, and ghrelin. For endogenous ghrelin, this led to an acyl/total ghrelin ratio of 1:5 instead of 1:19 by the standard method. These results show that the RAPID method enables accurate assessment of circulating gut peptide concentrations and forms such as CCK-58, acylated ghrelin, and somatostatin-28. Therefore, the RAPID method represents an efficacious means to detect circulating variations in peptide concentrations and form relevant to the understanding of physiological function of endocrine peptides.

Acknowledgments

We thank Dr. Stengel for giving us this precious information..

This procedure has the possibility to apply following our products;

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.

YK240 Corticosterone EIA

1.    What is Corticosterone?
Corticosterone (C21H30O4, also called 11beta, 21-Dihydroxyprogesterone, Reichstein’s Substance H, or Kendall’s Compound B) is, like cortisol and cortisone, a glucocorticoid hormone secreted from the cortex of adrenal gland.
It is a primary glucocorticoid in mice and rats and other animals (such as rabbits, birds, amphibians, and reptilians) in which the 17alfa-hydroxylase is supposed not to exist in adrenal gland.
Corticosterone is produced under the control of ACTH and the production has a circadian rhythm with peak levels in the latter portion of the day in nocturnal animals like rats and is believed to play a decisive role in sleep-wake cycle. Corticosterone can be used as a non-invasive biomarker of stress study through the collection of urine and feces to avoid corticosterone increase of blood levels which is caused by normal invasive methods.

2.    Specification
YK240 Corticosterone EIA kit is a quite useful tool for further development of corticosterone research.
This assay kit is newly developed by our laboratory provides a tool for direct determination of corticosterone in blood by simple dilution of blood samples with the diluent included in the kit. Any complicated extraction processes in sample preparation like other commercial kit are not needed. Furthermore, assays using the kit can be completed within a short period.

Cross reactivities of the antibody used in the kit.

Compound	Cross Reactivity （％）
Corticosterone	100
11-Deoxycorticosterone	<15.5
Progesterone	<5.9
Androstenedione	<5.4
Testosterone	<3.9
Aldosterone	<1.7
Cortisol	<0.5
Cortisone	<0.4
DHEA	<0.05
Estradiol	0
Cholesterol	0

3.    Assay principal
This EIA kit for determination of corticosterone is based on a competitive enzyme immunoassay using combination of specific antibody to corticosterone and corticosterone- horseradish peroxidase (HRP) conjugate (HRP-labeled corticosterone) system.


4.    Kit contents

5.    Technical data

Assay range is 0.21-50 ng/mL.
For detail, see package insert. Clock here to package insert.

6.    Research field

-    Circadian rhythm for mice and rats and other animals (such as rabbits, birds, amphibians, and reptilians).
-    As urinary stress marker.
-    Impairment of long-term memory retrieval.
-    Dietary restrictions.
-    Burn injuries, etc.

7.    Product description

Click here to jump to product description

Correlation after improvement (n=44)
correlation　r=0.960　
Regression analysis　Y=0.544X － 0.091　
Y=the measurement value after improvement
X=the measurement value before improvement



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September 17, 2009

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