Human PRSS1 (Protease, Serine, 1) CLIA Kit (HUES00684)

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SKU:
HUES00684
€599

Description

ELISA Kit Technical ManualMSDS

Human PRSS1 (Protease, Serine, 1) CLIA Kit

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 typeSandwich
Format96T
Assay time4.5h
ReactivityHuman
Detection methodChemiluminescence
Detection range0.16-10 ng/mL
Sensitivity0.09 ng/mL
Sample volume100uL
Sample typeSerum, plasma and other biological fluids
RepeatabilityCV < 15%

Specificity

This kit recognizes Human PRSS1 in samples. No significant cross-reactivity or interference between Human PRSS1 and analogues was observed.

Typical Data

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
1031717
33225
3247132447
514167
16773
1547015446
2.57959
7047
75037479
1.253601
3705
36533629
0.631860
1662
17611737
0.31866
782
824800
0.16334
380
357333
023
25
24--

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, mid range and high level Human PRSS1 were tested 20 times on one plate, respectively.

Inter-assay Precision (Precision between assays): 3 samples with low, mid range and high level Human PRSS1 were tested on 3 different plates, 20 replicates in each plate.

  Intra-assay Precision Inter-assay Precision
Sample123123
n202020202020
Mean (ng/mL)0.471.254.130.441.364.34
Standard deviation0.050.100.360.050.110.32
C V (%)10.648.008.7211.368.097.37

Recovery

The recovery of Human PRSS1 spiked 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)90-10196
EDTA plasma (n=5)91-10296
Cell culture media (n=5)96-110102

Linearity

Samples were spiked with high concentrations of Human PRSS1 and 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)
1:2Range (%)93-10694-106103-117
Average (%)98100110
1:4Range (%)86-10095-10897-109
Average (%)92101102
1:8Range (%)93-10690-10290-102
Average (%)1009595
1:16Range (%)97-11093-10796-107
Average (%)105100101

Kit Components & Storage

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 Standard2 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 Diluent1 vial, 20 mL4'C, 6 months
Biotinylated Detection Ab Diluent1 vial, 14 mL
HRP Conjugate Diluent1 vial, 14 mL
Concentrated Wash Buffer (25X)1 vial, 30 mL
Substrate Reagent A1 vial, 5 mL4'C (shading light)
Substrate Reagent B1 vial, 5 mL4'C (shading light)
Plate Sealer5 pieces
Product Description1 copy
Certificate of Analysis1 copy

Human PRSS1 (Protease, Serine, 1) CLIA Kit (HUES00684) Assay procedure

    1. Set standard, test sample and control (zero) wells on the pre-coated plate and record theirpositions. It is recommended to measure each standard and sample in duplicate. Note: addall solutions to the bottom of the plate wells while avoiding contact with the well walls. Ensuresolutions do not foam when adding to the wells.
    2. Aliquot 100µl of standard solutions into the standard wells.
    3. Add 100µl of Sample / Standard dilution buffer into the control (zero) well.
    4. Add 100µl of properly diluted sample (serum, plasma, tissue homogenates and otherbiological fluids.) into test sample wells.
    5. Cover the plate with the sealer provided in the kit and incubate for 90 min at 37°C.
    6. Aspirate the liquid from each well, do not wash. Immediately add 100µL of BiotinylatedDetection Ab working solution to each well. Cover the plate with a plate seal and gently mix.Incubate for 1 hour at 37°C.
    7. Aspirate or decant the solution from the plate and add 350µL of wash buffer to each welland incubate for 1-2 minutes at room temperature. Aspirate the solution from each well andclap the plate on absorbent filter paper to dry. Repeat this process 3 times. Note: a microplatewasher can be used in this step and other wash steps.
    8. Add 100µL of HRP Conjugate working solution to each well. Cover with a plate seal andincubate for 30 min at 37°C.
    9. Aspirate or decant the solution from each well. Repeat the wash process for five times asconducted in step 7.
    10. Add 100µL of Substrate mixture solution to each well. Cover with a new plate seal andincubate for no more than 5 min at 37°C. Protect the plate from light.
    11. Determine the RLU value of each well immediately.

Human PRSS1 (Protease, Serine, 1) CLIA Kit (HUES00684) Protein Information

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
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Additional Information

Reactivity:
Human
ELISA Type:
CLIA
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