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Cat Calprotectin (CALPRO) ELISA Kit


Species Reactivity

Cat (Felis catus,Feline)

UniProt

N/A

Abbreviation

CALPRO

Alternative Names

CAL; MRP-8/MRP-14; S100A8/A9

Range

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Sensitivity

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Sample Type

Serum, Plasma, Other biological fluids

Detection Method

Sandwich

Analysis Method

Quantitive

Assay Duration

1-4.5h

Sample Volume

1-200 μL

Detection Wavelengt

450 nm

Cat

AE64533CA

Size

48T 96T

Price

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Availability

3-5 working days

Manual

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Test principle:
This assay employs a two-site sandwich ELISA to quantitate CALPRO in samples. An antibody specific for CALPRO has been pre-coated onto a microplate. Standards and samples are pipetted into the wells and any CALPRO present is bound by the immobilized antibody. After removing any unbound substances, a biotin-conjugated antibody specific for CALPRO is added to the wells. After washing, Streptavidin conjugated Horseradish Peroxidase (HRP) is added to the wells. Following a wash to remove any unbound avidin-enzyme reagent, a substrate solution is added to the wells and color develops in proportion to the amount of CALPRO bound in the initial step. The color development is stopped and the intensity of the color is measured.

Product Overview:
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.

Components:

Reagents

Quantity

Reagents

Quantity

Assay plate (96 Wells)

1

Instruction manual

1

Standard (lyophilized)

2

Sample Diluent

1 x 20 mL

Biotin-Conjugate (concentrate 100 x)

1 x 120 μL

Biotin-Conjugate Diluent

1 x 20 mL

Streptavidin-HRP  (concentrate 100 x)

1 x 120 μL

Streptavidin-HRP Diluent   

1 x 20 mL

Wash Buffer (concentrate 25 x)

1 x 20 mL

Substrate Solution

1 x 12 mL

Stop Solution

1 x 10 mL

Adhesive Films

4

Specificity:
This assay has high sensitivity and excellent specificity for detection of Cat CALPRO. No significant cross-reactivity or interference between Cat CALPRO and analogues was observed.
Recovery:
Matrices listed below were spiked with certain level of recombinant Cat CALPRO and the recovery rates were calculated by comparing the measured value to the expected amount of Cat CALPRO in samples.

Sample Type

Number

Recovery range (%)

Average(%)

Serum

10

90-101

96

EDTA plasma

10

89-97

93

Heparin plasma

10

91-99

95

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

Sample Type

1:2

1:4

1:8

1:16

Serum

78-89%

81-99%

92-103%

95-105%

EDTA plasma

91-101%

90-98%

93-101%

91-98%

Heparin plasma

92-103%

93-102%

92-99%

91-101%

Stability:
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).
Sample collection and storage:
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.
Kits storage instructions:
Store at 2-8°C. Please refer to Instruction Manual.