ELISA Kit for Tryptase (TPS)

TPSAB1; Tryptase Alpha/Beta 1; Tryptase Alpha II; Tryptase Beta I; Tryptase-I; Tryptase-II; Tryptase-III

Specificity

This assay has high sensitivity and excellent specificity for detection of Tryptase (TPS).
No significant cross-reactivity or interference between Tryptase (TPS) and analogues was observed.

Recovery

Matrices listed below were spiked with certain level of recombinant Tryptase (TPS) and the recovery rates were calculated by comparing the measured value to the expected amount of Tryptase (TPS) in samples.

Matrix Recovery range (%) Average(%)
serum(n=5) 92-99 96
EDTA plasma(n=5) 82-93 86
heparin plasma(n=5) 97-105 102

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Tryptase (TPS) were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level Tryptase (TPS) were tested on 3 different plates, 8 replicates in each plate.
CV(%) = SD/meanX100
Intra-Assay: CV<10%
Inter-Assay: CV<12%

Linearity

The linearity of the kit was assayed by testing samples spiked with appropriate concentration of Tryptase (TPS) and their serial dilutions. The results were demonstrated by the percentage of calculated concentration to the expected.

Sample 1:2 1:4 1:8 1:16
serum(n=5) 98-105% 81-105% 92-102% 80-90%
EDTA plasma(n=5) 91-103% 87-101% 85-103% 80-101%
heparin plasma(n=5) 85-103% 87-96% 90-104% 98-105%

Stability

The stability of kit is determined by the loss rate of activity. The loss rate of this kit is less than 5% within the expiration date under appropriate storage condition.
To minimize extra influence on the performance, operation procedures and lab conditions, especially room temperature, air humidity, incubator temperature should be strictly controlled. It is also strongly suggested that the whole assay is performed by the same operator from the beginning to the end.

Reagents and materials provided

Reagents Quantity Reagents Quantity
Pre-coated, ready to use 96-well strip plate 1 Plate sealer for 96 wells 4
Standard 2 Standard Diluent 1×20mL
Detection Reagent A 1×120µL Assay Diluent A 1×12mL
Detection Reagent B 1×120µL Assay Diluent B 1×12mL
TMB Substrate 1×9mL Stop Solution 1×6mL
Wash Buffer (30 × concentrate) 1×20mL Instruction manual 1

Assay procedure summary

1. Prepare all reagents, samples and standards;
2. Add 100µL standard or sample to each well. Incubate 1 hours at 37°C;
3. Aspirate and add 100µL prepared Detection Reagent A. Incubate 1 hour at 37°C;
4. Aspirate and wash 3 times;
5. Add 100µL prepared Detection Reagent B. Incubate 30 minutes at 37°C;
6. Aspirate and wash 5 times;
7. Add 90µL Substrate Solution. Incubate 10-20 minutes at 37°C;
8. Add 50µL Stop Solution. Read at 450nm immediately.

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Magazine Citations
Journal of Allergy and Clinical Immunology Glandular mast cells with distinct phenotype are highly elevated in chronic rhinosinusitis with nasal polyps PubMed: 22534535
J Pediatr. Improved liver function and relieved pruritus after 4-phenylbutyrate therapy in a patient with progressive familial intrahepatic cholestasis type 2 Pubmed:24530123
Mediators of Inflammation Mediators of Mast Cells in Bullous Pemphigoid and Dermatitis Herpetiformis Pubmed:25400334
PLOS ONE A Novel Model of IgE-Mediated Passive Pulmonary Anaphylaxis in Rats Pubmed:25541997
Neurogastroenterology &amp; Motility Peritoneal mast cell degranulation and gastrointestinal recovery in patients undergoing colorectal surgery Pubmed:25677271
Mediators Inflamm Elevated Serum Tryptase and Endothelin in Patients with ST Segment Elevation Myocardial Infarction: Preliminary Report PubMed: 26089601
Int Immunopharmacol Tryptase and protease-activated receptor-2 stimulate scratching behavior in a murine model of ovalbumin-induced atopic-like dermatitis PubMed: 26049029
Mediators of Inflammation Altered expression of IFN-λ2 in allergic airway disorders and identification of its cell origins Journals: Mi
Mediators of Inflammation Altered Expression of IFN-2 in Allergic Airway Disorders and Identification of Its Cell Origins Pubmed:27057098
Analytica Chimica Acta An ultrasensitive electrochemical detection of tryptase using 3D macroporous reduced graphene oxide nanocomposites by one-pot electrochemical synthesis
Journal of Clinical Investigation Dengue virus-elicited tryptase induces endothelial permeability and shock Pubmed: 31265436
Frontiers in Physiology Mast Cells in Peritoneal Fluid From Women With Endometriosis and Their Possible Role in Modulating Sperm Function Pubmed: 31998139
Biomarkers of Allergic Asthma and their Association with Serum Parameters
Annals of Tropical Medicine & Public Health Effectiveness of biomarkers and serum parameters in determination allergic asthma and detection of its severity
Front Immunol Serum CD203c+ Extracellular Vesicle Serves as a Novel Diagnostic and Prognostic Biomarker for Succinylated Gelatin Induced Perioperative Hypersensitive … 34650557
Microbiol Spectr Mast Cell-Derived Proteases Induce Endothelial Permeability and Vascular Damage in Severe Fever with Thrombocytopenia Syndrome Pubmed:35612327
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