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Multiplex Assay Kit for Calprotectin (CALPRO) ,etc. by FLIA (Flow Luminescence Immunoassay)

MRP-8/MRP-14; S100A8/A9

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

Specificity

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

Recovery

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

Matrix Recovery range (%) Average(%)
serum(n=5) 78-99 88
EDTA plasma(n=5) 93-105 102
heparin plasma(n=5) 78-98 91
sodium citrate plasma(n=5) 78-102 85

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Calprotectin (CALPRO) ,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 Calprotectin (CALPRO) ,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 Calprotectin (CALPRO) ,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) 96-104% 88-96% 81-95% 95-103%
EDTA plasma(n=5) 81-93% 82-89% 91-101% 85-94%
heparin plasma(n=5) 94-105% 85-104% 92-101% 91-101%
sodium citrate plasma(n=5) 98-105% 97-105% 87-101% 83-103%

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#:CALPRO) 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
Medicine (Baltimore) A Novel Combination of Calprotectin and CXCL12 for Predicting Malignancy in Patients with Exudative Pleural Effusion PubMed: 26632726
Neurochemical Research  Effects of (-)-Sesamin on Chronic Stress-Induced Anxiety Disorders in Mice pubmed:27995494
International journal of molecular sciences Immune Modulating Topical S100A8/A9 Inhibits Growth of Pseudomonas aeruginosa and Mitigates Biofilm Infection in Chronic Wounds. pubmed:28672877
Analytical Chemistry  Multiplex Immuno-MALDI-TOF MS for Targeted Quantification of Protein Biomarkers and Their Proteoforms Related to Inflammation and Renal Dysfunction Pubmed:29420882
 JOURNAL OF OBSTETRICS AND GYNAECOLOGY RESEARCH Maternal serum calprotectin level in intrahepatic cholestasis of pregnancy 34254410
Bull Exp Biol Med Influence of Rectal Ozone Application on the Intensity of Free Radical Destruction of Lipids and Intestinal Proteins in the Dynamics of Experimental Colitis Pubmed:35622259
Serum Calprotectin and Iron Metabolism Biomarker Levels in Behçet's Disease
Catalog No. Related products for research use of Rattus norvegicus (Rat) Organism species Applications (RESEARCH USE ONLY!)
SEK504Ra ELISA Kit for Calprotectin (CALPRO) Enzyme-linked immunosorbent assay for Antigen Detection.
LMK504Ra Multiplex Assay Kit for Calprotectin (CALPRO) ,etc. by FLIA (Flow Luminescence Immunoassay) FLIA Kit for Antigen Detection.