Multiplex Assay Kit for Caspase 1 (CASP1) ,etc. by FLIA (Flow Luminescence Immunoassay)

ICE; IL1BC; IL1-BC; P45; Interleukin 1 Beta Converting Enzyme; Apoptosis-Related Cysteine Peptidase; Interleukin-1 beta convertase

(Note: Up to 8-plex in one testing reaction)

Specificity

This assay has high sensitivity and excellent specificity for detection of Caspase 1 (CASP1) ,etc. by FLIA (Flow Luminescence Immunoassay).
No significant cross-reactivity or interference between Caspase 1 (CASP1) ,etc. by FLIA (Flow Luminescence Immunoassay) and analogues was observed.

Recovery

Matrices listed below were spiked with certain level of recombinant Caspase 1 (CASP1) ,etc. by FLIA (Flow Luminescence Immunoassay) and the recovery rates were calculated by comparing the measured value to the expected amount of Caspase 1 (CASP1) ,etc. by FLIA (Flow Luminescence Immunoassay) in samples.

Matrix Recovery range (%) Average(%)
serum(n=5) 78-91 86
EDTA plasma(n=5) 79-89 80
heparin plasma(n=5) 80-94 85
sodium citrate plasma(n=5) 88-102 94

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Caspase 1 (CASP1) ,etc. by FLIA (Flow Luminescence Immunoassay) were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level Caspase 1 (CASP1) ,etc. by FLIA (Flow Luminescence Immunoassay) 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 Caspase 1 (CASP1) ,etc. by FLIA (Flow Luminescence Immunoassay) 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) 92-105% 79-94% 93-101% 84-103%
EDTA plasma(n=5) 87-101% 96-103% 97-104% 83-94%
heparin plasma(n=5) 98-105% 91-105% 96-103% 92-99%
sodium citrate plasma(n=5) 99-105% 90-104% 82-104% 92-105%

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
96-well plate 1 Plate sealer for 96 wells 4
Pre-Mixed Standard 2 Standard Diluent 1×20mL
Pre-Mixed Magnetic beads (22#:CASP1) 1 Analysis buffer 1×20mL
Pre-Mixed Detection Reagent A 1×120μL Assay Diluent A 1×12mL
Detection Reagent B (PE-SA) 1×120μL Assay Diluent B 1×12mL
Sheath Fluid 1×10mL Wash Buffer (30 × concentrate) 1×20mL
Instruction manual 1

Assay procedure summary

1. Preparation of standards, reagents and samples before the experiment;
2. Add 100μL standard or sample to each well,
    add 10μL magnetic beads, and incubate 90min at 37°C on shaker;
3. Remove liquid on magnetic frame, add 100μL prepared Detection Reagent A. Incubate 60min at 37°C on shaker;
4. Wash plate on magnetic frame for three times;
5. Add 100μL prepared Detection Reagent B, and incubate 30 min at 37°C on shaker;
6. Wash plate on magnetic frame for three times;
7. Add 100μL sheath solution, swirl for 2 minutes, read on the machine.

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Magazine Citations
PLoS One. Cilostazol Renoprotective Effect: Modulation of PPAR-γ, NGAL, KIM-1 and IL-18 Underlies Its Novel Effect in a Model of Ischemia-Reperfusion Pubmed:24816434
Fertil Steril Inflammosome in the human endometrium: further step in the evaluation of the “maternal side” PubMed: 26474737
PLoS One Synthetic PreImplantation Factor (PIF) prevents fetal loss by modulating LPS induced inflammatory response. pubmed:28704412
Journal of Virology NS1 protein of 2009 pandemic influenza A virus inhibits porcine NLRP3 inflammasome-mediated interleukin-1 beta production by suppressing ASC ubiquitination Pubmed:29386291
Journal of Translational Medicine Recurrent pregnancy loss is associated to leaky gut: a novel pathogenic model of endometrium inflammation? Pubmed:29665864
Reproductive Sciences A role for the inflammasome in spontaneous labor at term Pubmed:27852921
Journal of Virology CHARACTERIZATION OF PORCINE NLRP3 INFLAMMASOME-MEDIATED INTERLEUKIN-1 BETA PRODUCTION IN RESPONSE TO INFLUENZA A VIRUSES Pubmed: 29386291
ACTA Otorhinolaryngologica Italica NLRP3 inflammasome expression in lipopolysaccharide-induced otitis media Pubmed: 30676848
American Journal of Reproductive Immunology Effect of alpha lipoic acid and myoinositol on endometrial inflammasome from recurrent pregnancy loss women Pubmed: 31148259
Taiwanese Journal of Obstetrics & Gynecology Serum Caspase-1 levels in women with polycystic ovary syndrome Pubmed: 32127139
Iranian Journal of Allergy, Asthma and Immunology Increased Level of Caspase-1 in the Serum of Relapsing-remitting Multiple Sclerosis (RRMS) Patients
ACTIVITY OF APOPTOSIS MARKERS IN HPV-ASSOCIATED CERVICAL PATHOLOGIES
Norwegian Journal of development of the International Science CLINICAL AND PATHOGENETIC SIGNIFICANCE OF CHANGES IN PURINE METABOLISM DURING LUNG CANCER
LIFE SCIENCES The NLRP3 inhibitor dapansutrile attenuates folic acid induced nephrotoxicity via inhibiting inflammasome/caspase-1/IL axis and regulating autophagy/proliferation 34560082
Biol Reprod The alarmin S100A12 causes sterile inflammation of the human chorioamniotic membranes and preterm birth and neonatal mortality in mice 34632484
J Mol Neurosci TRPV1 Suppressed NLRP3 Through Regulating Autophagy in Microglia After Ischemia-Reperfusion Injury Pubmed:35041191
Viruses Influenza A Virus Infection Activates NLRP3 Inflammasome through Trans-Golgi Network Dispersion Pubmed:35062292
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