Rat/mouse OCT assay kit

Introduction

Non-alcoholic steatohepatitis (NASH) and alcoholic liver disease (ALD) show similar pathological findings, with the involvement of mitochondrial disorders1). Therefore, Yamasa Corporation investigated ornithine carbamoyltransferase (OCT), localized in liver mitochondria, as a marker of liver mitochondrial disorders, and developed rat/mouse OCT quantitative ELISA using a monoclonal antibody. Blood OCT concentrations in rat models of liver injury were examined with the ELISA, demonstrating that OCT was more useful than cytoplasmic markers, alanine aminotransferase (ALT) and aspartate aminotransferase (AST), in rat models of drug-induced liver injury 2)3), NASH 4) and ALD4).

A rat/mouse OCT assay kit, developed based on the ELISA, can be employed for quantifying rat and mouse serum OCT concentrations.

Features of the kit

– ELISA with a 96-well microtiter plate.
– Measurement time: primary reaction, 1 hour; secondary reaction, 2 hours; and color reaction, 30 minutes
– Rat and mouse*1 serum OCT concentrations can be measured.*2
*1: Rat OCT concentrations were used as standards. Mouse OCT concentrations were calculated as equivalents to the rat OCT concentrations.
*2: Samples need to be diluted before measurement. For details, see (2) of “Notes of measurement.”

Principles of measurement

Sandwich ELSIA was developed using two kinds of mouse anti-OCT monoclonal antibody.

Product contents

– Product code: 80127
– Components:

Name Volum Quantity
(1) Antibody Coated Plate 96-well microwells plate 1 plate
(2) OCT Standard 1 (0ng/mL) 0.05mL 1 vial
(3) OCT Standard 2 (30ng/mL) 0.05mL 1 vial
(4) OCT Standard 3 (100ng/mL) 0.05mL 1 vial
(5) OCT Standard 4 (300ng/mL) 0.05mL 1 vial
(6) Diluent 50mL 1 vial
(7) Concentrated Washing Solution 50mL 1 vial
(8) Enzyme Conjugate 0.15mL 1 vial
(9) Color Developing Reagent A 6mL 1 vial
(10) Color Developing Reagent B 6mL 1 vial
(11) Stop Solution 11mL 1 vial

– Storage: 2-8℃
– Shelf life: 6 months after the date of manufacture
– Suggested retail price: ¥ 143,000

Measuring methods

<Outline>

Dispense 100 μL of OCT Standard 1-4 or sample to Antibody Coated Plate.
↓ Incubate the plate for 1 hour at 20-30℃.
After removal of the reaction solution, dispense 300 μL of wash solution into each well.
↓ Repeat these procedures three times.
After removal of the reaction solution, dispense 100 μL of diluted Enzyme Conjugate to each well.
↓ Incubate the plate for 1 hour at 20-30℃.
After removal of the reaction solution, dispense 300 μL of washing solution into each well.
↓ Repeat the above procedures three times.
Dispense 100 μL of coloring solution into each well.
↓ Incubate the plate at 20 – 30 ℃ for 30 min.
Dispense 100 μL of stop solution to each well.
↓
Measure absorbance of each well at 450 nm.

Notes of measurement

(1) Serums of rat or muse are diluted 11-fold with Diluent.  If the OCT level of the sample exceed measuring range, dilute with Diluent to obtain a value within the range..
(2) For more information, contact Marketing & MR Section.

References

1. Pessayre D. et al. (2005). J Hepatol. 42. 928-40.
2. Murayama H. et al. (2007). Clin Chim Acta. 375. 63-68.
3. Murayama H. et al. (2008). Clin Chim Acta. 391. 31-35.
4. Murayama H. et al. :The 36th Annual Meeting of The Japanese Society of Toxicology
5. Furihata T et al. (2016). Drug Metab Pharmacokinet. 31. 102-105.