CLIA Kit for Calprotectin (CALPRO)

MRP-8/MRP-14; S100A8/A9

Specificity

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

Recovery

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

Matrix Recovery range (%) Average(%)
serum(n=5) 84-98 92
EDTA plasma(n=5) 95-103 99
heparin plasma(n=5) 80-94 89

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Calprotectin (CALPRO) were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level Calprotectin (CALPRO) 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) 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) 80-99% 93-101% 86-98% 81-101%
EDTA plasma(n=5) 82-96% 95-103% 85-104% 81-91%
heparin plasma(n=5) 93-101% 80-97% 99-105% 80-93%

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
Substrate A 1×10mL Substrate B 1×2mL
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 100µL Substrate Solution. Incubate 10 minutes at 37°C;
8. Read RLU value immediately.

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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 Canis familiaris; Canine (Dog) Organism species Applications (RESEARCH USE ONLY!)
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