Alanine Aminotransferase 1 ELISA Kit (GPT)

INTENDED USE

Human alanine aminotransferase 1 ELISA kit can measure concentrations of GPT (alanine aminotransferase 1, AAT1, Glutamic pyruvate transaminase) present is plasma, serum, biological fluids or tissue homogenate samples.

INTRODUCTION

Alanine aminotransferase 1 (ALT1), more commonly termed glutamate pyruvate transaminase (GPT), serves an essential role in glucose and amino acid metabolism as a hub for nitrogen shuttling between different compounds. It catalyzes transfer of an amino group from alanine to α-ketoglutarate, yielding pyruvate and glutamate products. First described in 1958, ALT became recognized as a liver-specific enzyme vital for gluconeogenesis and nitrogen homeostasis. During fasting, increases in ALT flux directionally promotes conversion of pyruvate derived from muscle-liberated alanine into glucose within the liver.

However, liver injury invariably causes large spills of ALT1 into circulation making it a routinely used clinical biomarker. Interestingly, baseline circulating ALT levels independently predict future risk for insulin resistance, metabolic syndrome, and type II diabetes mellitus. Minor ALT elevations likely reflect underlying hepatic lipid accumulation, the hallmark of non-alcoholic fatty liver disease now reaching epidemic prevalence.

Mechanistically, inflammatory and metabolic stresses trigger ALT1 release via microvesicle shedding and caspase-mediated cleavage. Once in plasma, it retains catalytic activity and binding partners from its intrinsic metabolic pathways. This combination of abundant liberation from damaged tissue coupled with endogenous enzyme function makes ALT1 a unique multi-purpose damage associated molecular pattern (DAMP) signal. Circulating ALT1-metabolites help sound systemic alarms through TLR4 immune activation while also directing metabolic adaptations in extra-hepatic organs. This unique capacity as both enzyme and DAMP underlies why modest ALT fluctuations can presage metabolic disease years later.

CONTENT

All reagents supplied need to be stored at 2 °C – 8 °C, unopened reagents will retain reactivity until expiration date. Do not use reagents beyond this date.

• One 96-Well Plate: Pre-coated with anti- GPT.
• Standards: Lyophilized recombinant.
• Sample/Standard Dilution Buffer.
• Biotinylated-labelled Antibody.
• Antibody Dilution Buffer.
• HRP-Streptavidin Conjugate (SABC).
• SABC Dilution Buffer.
• TMB Substrate.
• Wash Buffer (25x).
• Plate Sealer.
• Product Instructions.

TYPICAL RESULTS

For this GPT ELISA kit, it is recommended that a standard curve is generated for each assay carried out.

Standard Curve: 0, 3.125, 6.25, 12.5, 25, 50, 100, 200 mIU/ml
Reactivity: Human
Sensitivity: 1.875 mIU/ml
Range: 3.125 – 200 mIU/ml
Principle: Sandwich, Double Antibody
Application: Research Use Only.

ASSAY CHARACTERISTICS

– Specificity: Highly specific for GPT, no cross reactivity or interference between GPT and analogues was detected.
– Recovery: Serum (86 – 102%), EDTA Plasma (94 – 101%), Heparin Plasma (92 – 103%).
– Linearity: Serum (87 – 103%), EDTA Plasma (83 – 100%), Heparin Plasma (81 – 100%).
– Precison Intra-Assay: CV < 8%
– Precison Inter-Assay: CV < 10%

REFERENCES

1. On elevated alanine aminotransferase in type 1 diabetes mellitus. Minerva Gastroenterol Dietol. (2016) 62 (2): 223. Pellicano R. and Saracco G.M.
2. Enzymes involved in l-lactate metabolism in humans. Mitochondrion. (2013) 13 (6): 615-29. Adeva M., et al.
3. Alanine aminotransferase in clinical practice. Arch Intern Med. (1992) 152 (1): 208. Hodgson M.J.

ADDITIONAL INFORMATION

• Full Name: Alanine Aminotransferase 1 ELISA Kit (GPT)
• Reactivity: Human
• Sample Type: Plasma, Tissue Homogenates, Serum, Biological Fluids
• Sensitivity: 1.875 mIU/ml

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