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This kit uses Sandwich-CLIA as the method. The micro CLIA plate provided in this kit has been pre-coated with an antibody specific to Human PRSS1 . Standards or samples are added to the appropriate micro CLIA plate wells and combined with the specific antibody. Then a biotinylated detection antibody specific for Human PRSS1 and Avidin-Horseradish Peroxidase (HRP) conjugate are added to each micro plate well successively and incubated. Free components are washed away. The substrate solution is added to each well. Only those wells that contain Human PRSS1, biotinylated detection antibody and Avidin-HRP conjugate will appear fluorescence. The Relative light unit (RLU) value is measured spectrophotometrically by the Chemiluminescence immunoassay analyzer. The RLU value is positively associated with the concentration of Human PRSS1. The concentration of Human PRSS1 in the samples can be calculated by comparing the RLU of the samples to the standard curve.
| Assay type | Sandwich |
| Format | 96T |
| Assay time | 4.5h |
| Reactivity | Human |
| Detection method | Chemiluminescence |
| Detection range | 0.16-10 ng/mL |
| Sensitivity | 0.09 ng/mL |
| Sample volume | 100uL |
| Sample type | Serum, plasma and other biological fluids |
| Repeatability | CV < 15% |
This kit recognizes Human PRSS1in samples. No significant cross-reactivity or interference between Human PRSS1and analogues was observed.
As the RLU values of the standard curve may vary according to the conditions of the actual assay performance (e.g. operator, pipetting technique, washing technique or temperature effects), the operator should establish a standard curve for each test. Typical standard curve and data is provided below for reference only.
| Concentration(ng/mL) | RLU | Average | Corrected |
|---|---|---|---|
| 10 | 3171733225 | 32471 | 32447 |
| 5 | 1416716773 | 15470 | 15446 |
| 2.5 | 79597047 | 7503 | 7479 |
| 1.25 | 36013705 | 3653 | 3629 |
| 0.63 | 18601662 | 1761 | 1737 |
| 0.31 | 866782 | 824 | 800 |
| 0.16 | 334380 | 357 | 333 |
| 0 | 2325 | 24 | -- |
Intra-assay Precision (Precision within an assay): 3 samples with low, mid range and high level Human PRSS1were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, mid range and high level Human PRSS1were tested on 3 different plates, 20 replicates in each plate.
| Intra-assay Precision | Inter-assay Precision | |||||
|---|---|---|---|---|---|---|
| Sample | 1 | 2 | 3 | 1 | 2 | 3 |
| n | 20 | 20 | 20 | 20 | 20 | 20 |
| Mean (ng/mL) | 0.47 | 1.25 | 4.13 | 0.44 | 1.36 | 4.34 |
| Standard deviation | 0.05 | 0.10 | 0.36 | 0.05 | 0.11 | 0.32 |
| C V (%) | 10.64 | 8.00 | 8.72 | 11.36 | 8.09 | 7.37 |
The recovery of Human PRSS1spiked at three different levels in samples throughout the range of the assay was evaluated in various matrices.
| Sample Type | Range (%) | Average Recovery (%) |
|---|---|---|
| Serum (n=5) | 90-101 | 96 |
| EDTA plasma (n=5) | 91-102 | 96 |
| Cell culture media (n=5) | 96-110 | 102 |
Samples were spiked with high concentrations of Human PRSS1and diluted with Reference Standard & Sample Diluent to produce samples with values within the range of the assay.
| Serum (n=5) | EDTA plasma (n=5) | Cell culture media (n=5) | ||
|---|---|---|---|---|
| 1:2 | Range (%) | 93-106 | 94-106 | 103-117 |
| Average (%) | 98 | 100 | 110 | |
| 1:4 | Range (%) | 86-100 | 95-108 | 97-109 |
| Average (%) | 92 | 101 | 102 | |
| 1:8 | Range (%) | 93-106 | 90-102 | 90-102 |
| Average (%) | 100 | 95 | 95 | |
| 1:16 | Range (%) | 97-110 | 93-107 | 96-107 |
| Average (%) | 105 | 100 | 101 |
An unopened kit can be stored at 4"C for 1 month. If the kit is not used within 1 month, store the items separately according to the following conditions once the kit is received.
| Item | Specifications | Storage |
|---|---|---|
| Micro CLIA Plate(Dismountable) | 8 wells X12 strips | -20"C, 6 months |
| Reference Standard | 2 vials | |
| Concentrated Biotinylated Detection Ab (100X) | 1 vial, 120 uL | |
| Concentrated HRP Conjugate (100X) | 1 vial, 120 uL | -20"C(shading light), 6 months |
| Reference Standard & Sample Diluent | 1 vial, 20 mL | 4"C, 6 months |
| Biotinylated Detection Ab Diluent | 1 vial, 14 mL | |
| HRP Conjugate Diluent | 1 vial, 14 mL | |
| Concentrated Wash Buffer (25X) | 1 vial, 30 mL | |
| Substrate Reagent A | 1 vial, 5 mL | 4"C (shading light) |
| Substrate Reagent B | 1 vial, 5 mL | 4"C (shading light) |
| Plate Sealer | 5 pieces | |
| Product Description | 1 copy | |
| Certificate of Analysis | 1 copy |
| UniProt Protein Function: | trypsin 1: Has activity against the synthetic substrates Boc-Phe- Ser-Arg-Mec, Boc-Leu-Thr-Arg-Mec, Boc-Gln-Ala-Arg-Mec and Boc-Val- Pro-Arg-Mec. The single-chain form is more active than the two- chain form against all of these substrates. Defects in PRSS1 are a cause of pancreatitis (PCTT). A disease characterized by the presence of calculi in pancreatic ducts. It causes severe abdominal pain attacks. Belongs to the peptidase S1 family. |
| UniProt Protein Details: | Protein type:Secreted, signal peptide; EC 3.4.21.4; Protease; Secreted Chromosomal Location of Human Ortholog: 7q34 Cellular Component: extracellular region Molecular Function:serine-type endopeptidase activity; metal ion binding Biological Process: extracellular matrix disassembly; extracellular matrix organization and biogenesis; vitamin metabolic process; cobalamin metabolic process; digestion; proteolysis; water-soluble vitamin metabolic process Disease: Trypsinogen Deficiency; Pancreatitis, Hereditary |
| NCBI Summary: | This gene encodes a trypsinogen, which is a member of the trypsin family of serine proteases. This enzyme is secreted by the pancreas and cleaved to its active form in the small intestine. It is active on peptide linkages involving the carboxyl group of lysine or arginine. Mutations in this gene are associated with hereditary pancreatitis. This gene and several other trypsinogen genes are localized to the T cell receptor beta locus on chromosome 7. [provided by RefSeq, Jul 2008] |
| UniProt Code: | P07477 |
| NCBI GenInfo Identifier: | 136408 |
| NCBI Gene ID: | 5644 |
| NCBI Accession: | P07477.1 |
| UniProt Secondary Accession: | P07477,Q5NV57, Q7M4N3, Q7M4N4, Q92955, Q9HAN4, Q9HAN5 Q9HAN6, Q9HAN7, A1A509, A6NJ71, B2R5I5, |
| UniProt Related Accession: | P07477 |
| Molecular Weight: | 26,558 Da |
| NCBI Full Name: | Trypsin-1 |
| NCBI Synonym Full Names: | protease, serine, 1 (trypsin 1) |
| NCBI Official Symbol: | PRSS1 |
| NCBI Official Synonym Symbols: | TRP1; TRY1; TRY4; TRYP1 |
| NCBI Protein Information: | trypsin-1; beta-trypsin; trypsinogen 1; trypsinogen A; digestive zymogen; cationic trypsinogen; nonfunctional trypsin 1 |
| UniProt Protein Name: | Trypsin-1 |
| UniProt Synonym Protein Names: | Beta-trypsin; Cationic trypsinogen; Serine protease 1; Trypsin ICleaved into the following 2 chains:Alpha-trypsin chain 1; Alpha-trypsin chain 2 |
| Protein Family: | Tripeptidyl aminopeptidase |
| UniProt Gene Name: | PRSS1 |
| UniProt Entry Name: | TRY1_HUMAN |
Reactivity: | Human |
ELISA Type: | CLIA |
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