Intended use 
This immunoassay kit allows for the in vitro quantitative determination of General cholyglycine concentrations in serum, Plasma, tissue homogenates and Cell culture supernates and Other biological fluids. 
Test principle 
The ELISA is based on the competitive binding enzyme immunoassay technique. The microtiter plate provided in this kit has been pre-coated with an antibody specific to General cholyglycine, During the reaction, General cholyglycine in the sample or standard competes with a fixed amount of biotin-labeled General cholyglycine for sites on a pre-coated Monoclonal antibody specific to General. Excess conjugate and unbound sample or standard are washed from the plate. Next, Avidin conjugated to Horseradish Peroxidase (HRP) is added to each microplate well and incubated. Then a TMB substrate solution is added to each well. The enzyme-substrate reaction is terminated by the addition of a sulphuric acid solution and the color change is measured spectrophotometrically at a wavelength of 450 nm ± 2 nm. The concentration of General cholyglycine in the samples is then determined by comparing the O.D. of the samples to the standard curve.

Linearity

The linearity of the kit was assayed by testing samples spiked with appropriate concentration of cholyglycine and their serial dilutions. The results were demonstrated by the percentage of calculated concentration to the expected.

Sample	1：2	1:4	1:8	1:16
serum(n=5)	111-121%	98-106%	106-116%	103-113%
EDTA plasma(n=5)	90-100%	100-112%	98-110%	105-115%
heparin plasma(n=5)	107-116%	105-115%	99-109%	97-106%

 

General Annotation

 

Function:

Serum cholyglycine (Cholyglycine, CG) is a combination of secondary bile acids with glycine conjugated bile acids, in liver cells, cholesterol, after its complex enzymatic reaction, turn into the primary bile acid. Cholic acid (CA) and chenodeoxycholic acid (the CD-CA). Bile acid steroid nucleus three hydroxyl groups (C3, C7, C12), the side chain hydroxyl at the end of the peptide bond and glycine combined with a molecular weight of 462u. CG normal metabolic pathways of the intestine - liver circulation, CG synthesized by hepatocytes, bile canaliculi, bile duct discharged into the gall bladder, along with the bile into the duodenum to help digestion of food. 95% of bile acid reabsorption in the distal ileum, come back to the liver via the portal vein, and uptake by the liver cells to re-use. Exist in the form of protein binding in serum, the total amount of overflow into the systemic circulation is less than 1%. Under normal circumstances, bile acid content in the peripheral blood, leaving little room for normal adults, whether fasting or postprandial serum CG concentration stabilized at a low level. When liver cells are damaged, the liver cell uptake CG capacity decreased, resulting in increased CG content in the blood; cholestasis, liver excretion of bile acids that barriers to reflux of blood circulation increased CG content, but also to the blood CG levels increased. Therefore, the determination of serum glycocholic acid (SCG) by ELISA method is a sensitive indicator to evaluate the function of liver cells and their hepatobiliary material circulation.

 

Location:

Normal serum cholyglycine: 1.3 ± 0.8mg / L, range 0.4 to 2.98mg / L, and the low limit of the hepatitis diagnosis <3.18mg / L

 

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