Porcine Trypsin ELISA Kit (Try)

Full Name: Porcine Trypsin ELISA Kit (Try)
Reactivity: Porcine
Sample Type: Serum, Tissue Homogenates, Biological Fluids, Plasma
Sensitivity:  0.938 ng/ml

INTRODUCTION

Trypsin (Try) cleaves proteins into smaller peptides, aiding in the absorption of nutrients such as amino acids. It is also used in research laboratories to cut proteins and peptides into specific sizes for further analysis. It functions by cleaving predominately arginine and lysine residues that are located at the carboxyl terminal. It is usually denatured with heat, alkali or acidic solutions to make it more active and then the enzyme can be stored in a solution or kept in its native form.

Trypsin enzyme cuts itself and its substrate between arginine residues. The other end of the substrate is cleaved on a phenylalanine residue that is located at the carboxyl terminal. Natively, this enzyme also cuts itself and its substrate between aspartic acid residues.

Trypsin deficiency can cause malabsorption of proteins and this can lead to pancreatitis. However, this is a rare condition, as trypsinogen is activated into trypsin in the small intestine and enterokinase deficiency is more common. Historically, it was prepared from casein, a milk protein. However, in modern times, it is produced from soy protein. It is classified as a serine protease.

One trypsin molecule cleaves at least two peptide bonds in the non-catalysed reaction between the active site and its substrate, resulting in two fragments: an amide and a carboxyl group. Trypsin cleaves peptide bonds, mainly in non-catalysed reactions with its substrate. This results in the release of a carboxyl group and an amide fragment. The amide fragment is non-functional because it has lost its amino group, but the carboxyl group can react with other molecules to form esters and amides.

INTENDED USE

Porcine trypsin ELISA kit can be used for detecting concentration of trypsin (Try) using porcine plasma, other biological fluids, serum and tissue homogenate samples.

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-trypsin antibody.
• 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 porcine trypsin ELISA kit it is recommended that a standard curve is generated for each assay carried out.

Standard Curve: 0, 1.562, 3.125, 6.25, 12.5, 25, 50, 100 ng/ml.
Reactivity: Porcine
Sensitivity: 0.938 ng/ml
Range: 1.563 – 100 ng/ml
Principle: Sandwich
Application: Research Use Only.

ASSAY CHARACTERISTICS

– Specificity: Highly specific for Try, no cross reactivity or interference between Try and analogues was detected.
– Recovery: Serum (89 – 101%), EDTA Plasma (86 – 105%), Heparin Plasma (85 – 105%).
– Linearity: Serum (88 – 97%), EDTA Plasma (89 – 104%), Heparin Plasma (86 – 101%).
– Precison Intra-Assay: CV < 8%.
– Precison Inter-Assay: CV < 10%.
– Stability: Less than 10%.

REFERENCES

1. Trypsin-promotes porcine deltacoronavirus mediating cell-to-cell fusion in a cell type-dependent manner. Emerg Microbes Infect. (2020) 9 (1): 457-468. Yang Y.L., et al.
2. Porcine trypsin: separation of the two polypeptide chains of the alpha-form and partial amino acid sequence. FEBS Lett. (1973) 32 (2): 223-6. Walker J.E., et al.
3. Expression, activation and characterization of porcine_trypsin in Pichia pastoris GS115. Protein Expr Purif. (2015) 114: 149-55. Shu M., et al.
4. High-gradient magnetic affinity separation of trypsin from porcine pancreatin. Biotechnol Bioeng. (2002) 79 (3): 301-13. Hubbuch J.J. and Thomas O.R.

OTHER RELATED ELISA KITS

• Presepsin ELISA Kit
• Neopterin ELISA Kit
• Glycosaminoglycan ELISA Kit