Human PRSS2 (Protease, Serine, 2) ELISA Kit (HUES02250)

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HUES02250
€539

Description

ELISA Kit Technical ManualMSDS

Human PRSS2 (Protease, Serine, 2) ELISA Kit

This ELISA kit uses Sandwich-ELISA as the method. The micro ELISA plate provided in this kit has been pre-coated with an antibody specific to Human PRSS2 . Standards or samples are added to the appropriate micro ELISA plate wells and combined with the specific antibody. Then a biotinylated detection antibody specific for Human PRSS2 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 PRSS2, biotinylated detection antibody and Avidin-HRP conjugate will appear blue in color. The enzyme-substrate reaction is terminated by adding Stop Solution and the color turns yellow. The optical density (OD) is measured spectrophotometrically at a wavelength of 450 nm ± 2 nm. The OD value is proportional to the concentration of Human PRSS2. The concentration of Human PRSS2 in samples can be calculated by comparing the OD of the samples to the standard curve.

Assay typeSandwich
Format96T
Assay time4.5h
ReactivityHuman
Detection MethodColormetric
Detection Range0.78-50 ng/mL
Sensitivity0.47 ng/mL
Sample Volume Required Per Well100uL
Sample TypeSerum, plasma and other biological fluids

Specificity

This kit recognizes Human PRSS2 in samples. No significant cross-reactivity or interference between Human PRSS2 and analogues was observed.

Typical Data

As the OD 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)
O.D Average Corrected
502.413
2.445
2.4292.339
251.559
1.617
1.5881.498
12.50.892
0.884
0.8880.798
6.250.459
0.487
0.4730.383
3.130.277
0.255
0.2660.176
1.570.191
0.183
0.1870.097
0.780.137
0.143
0.140.05
00.085
0.095
0.09--

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, mid range and high level Human PRSS2 were tested 20 times on one plate, respectively.

Inter-assay Precision (Precision between assays): 3 samples with low, mid range and high level Human PRSS2 were tested on 3 different plates, 20 replicates in each plate.

Intra-assay Precision Inter-assay Precision
Sample123123
n202020202020
Mean (ng/mL)2.577.3619.432.506.7921.25
Standard deviation0.140.370.780.170.331.13
C V (%)5.455.034.016.804.865.32

Recovery

The recovery of Human PRSS2 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)88-10193
EDTA plasma (n=5)89-10095
Cell culture media (n=5)85-9892

Linearity

Samples were spiked with high concentrations of Human PRSS2 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 (%)99-11496-11098-112
Average (%)105101103
1:4Range (%)87-9985-9787-99
Average (%)949192
1:8Range (%)90-10185-10089-103
Average (%)959195
1:16Range (%)88-10485-10083-96
Average (%)959288

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 ELISA 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 Reagent1 vial, 10 mL4'C(shading light)
Stop Solution1 vial, 10 mL4'C
Plate Sealer5 pieces
Product Description1 copy
Certificate of Analysis1 copy

Human PRSS2 (Protease, Serine, 2) ELISA Kit (HUES02250) 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 90µL of Substrate Reagent to each well. Cover with a new plate seal and incubate forapproximately 15 min at 37°C. Protect the plate from light. Note: the reaction time can beshortened or extended according to the actual color change, but not by more than 30min.
    11. Add 50 µL of Stop Solution to each well. Note: Adding the stop solution should be done inthe same order as the substrate solution.
    12. Determine the optical density (OD value) of each well immediately with a microplate readerset at 450 nm.

Human PRSS2 (Protease, Serine, 2) ELISA Kit (HUES02250) Protein Information

UniProt Protein Function:trypsin 2: In the ileum, may be involved in defensin processing, including DEFA5. Belongs to the peptidase S1 family.
UniProt Protein Details:

Protein type:Cell adhesion; Secreted, signal peptide; Calcium-binding; Secreted; EC 3.4.21.4; Extracellular matrix; Protease

Chromosomal Location of Human Ortholog: 7q34

Cellular Component: extracellular matrix; extracellular region; extracellular space

Molecular Function:calcium ion binding; protein binding; serine-type endopeptidase activity; serine-type peptidase activity

Biological Process: collagen catabolic process; digestion; extracellular matrix disassembly; positive regulation of cell adhesion; positive regulation of cell growth; proteolysis

Disease: Pancreatitis, Hereditary

NCBI Summary:This gene belongs to the trypsin family of serine proteases and encodes anionic trypsinogen. It is part of a cluster of trypsinogen genes that are located within the T cell receptor beta locus. Enzymes of this family cleave peptide bonds that follow lysine or arginine residues. This protein is found at high levels in pancreatic juice and its upregulation is a characteristic feature of pancreatitis. This protein has also been found to activate pro-urokinase in ovarian tumors, suggesting a function in tumor invasion. In addition, this enzyme is able to cleave across the type II collagen triple helix in rheumatoid arthritis synovitis tissue, potentially participating in the degradation of type II collagen-rich cartilage matrix. Alternative splicing results in multiple transcript variants.[provided by RefSeq, Jan 2015]
UniProt Code:P07478
NCBI GenInfo Identifier:136413
NCBI Gene ID:5645
NCBI Accession:P07478.1
UniProt Related Accession:P07478
Molecular Weight:26,488 Da
NCBI Full Name:Trypsin-2
NCBI Synonym Full Names:protease, serine 2
NCBI Official Symbol:PRSS2  
NCBI Official Synonym Symbols:TRY2; TRY8; TRYP2  
NCBI Protein Information:trypsin-2
UniProt Protein Name:Trypsin-2
UniProt Synonym Protein Names:Anionic trypsinogen; Serine protease 2; Trypsin II
Protein Family:Trypsin
UniProt Gene Name:PRSS2  
UniProt Entry Name:TRY2_HUMAN
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Additional Information

Reactivity:
Human
ELISA Type:
Sandwich
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