Bovine Calprotectin (CALPRO) ELISA Kit

Calprotectin is a protein belonging to the S100 family and occurring in large amounts in neutrophil granulocytes, where it accounts for 5% of total proteins and 60% of cytoplasm proteins. Smaller amounts of calprotectin has also been detected in monocytes and activated macrophages. The structure of calprotectin consists of a light polypeptide chain and two heavy polypeptide chains, with a molecular weight of 36,5 kDa. Bacteriostatic and mycostatic properties of calprotectin are comparable to those of antibiotics. For this reason the abundance of calprotectin in neutrophil granulocytes and its antimicrobial activity suggest a substantial role in the defence of the organism.Calprotectin has been found in several human biological materials: serum, saliva, cerebrospinal fluid and urine.

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

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

Intra-assay Precision (Precision within an assay) Three samples of known concentration were tested twenty times on one plate to assess intra-assay precision. Inter-assay Precision (Precision between assays) Three samples of known concentration were tested in forty separate assays to assess inter-assay precision. CV (%) = SD/meanX100 Intra-Assay: CV<8% Inter-Assay: CV<12%

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

The stability of ELISA 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. The loss rate was determined by accelerated thermal degradation test. Keep the kit at 37°C for 4 and 7 days, and compare O.D.values of the kit kept at 37°C with that of at recommended temperature. (referring from China Biological Products Standard, which was calculated by the Arrhenius equation. For ELISA kit, 4 days storage at 37°C can be considered as 6 months at 2 - 8°C, which means 7 days at 37°C equaling 12 months at 2 - 8°C).

Serum: Use a serum separator tube (SST) and allow samples to clot for two hours at room temperature or overnight at 2 - 8°C before centrifugation for 15 minutes at 1000 × g. Remove serum and assay immediately or aliquot and store samples at ≤ -20°C. Avoid repeated freeze-thaw cycles. Plasma: Collect plasma using EDTA, or heparin as an anticoagulant. Centrifuge for 15 minutes at 1000 × g at 2 - 8°C within 30 minutes of collection. Assay immediately or aliquot and store samples at ≤ -20°C. Avoid repeated freeze-thaw cycles. Other biological fluids: Centrifuge samples for 20 minutes at 1000 × g. Remove particulates and assay immediately or store samples in aliquot at -20°C or -80°C. Avoid repeated freeze/thaw cycles.