Horse Cystic fibrosis transmembrane conductance regulator (CFTR) ELISA Kit

SKU: AE50680HO

Size:96-test Kit. Other sizes are also available. Please contact us.

Species Reactivity	Horse (Equus caballus; Equine)
UniProt	Q2QLA3
Abbreviation	CFTR
Alternative Names	tcag7.78; ABC35; ABCC7; CF; CFTR/MRP; MRP7; TNR-CFTR; dJ760C5.1; ATP-binding cassette sub-family C; member 7|cystic fibrosis transmembrane conductance regulator
Range	Request Information
Sensitivity	Request Information
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

Test principle

This assay employs a two-site sandwich ELISA to quantitate CFTR in samples. An antibody specific for CFTR has been pre-coated onto a microplate. Standards and samples are pipetted into the wells and any CFTR present is bound by the immobilized antibody. After removing any unbound substances, a biotin-conjugated antibody specific for CFTR 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 CFTR bound in the initial step. The color development is stopped and the intensity of the color is measured.

Product Overview

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 12 mL
Streptavidin-HRP (concentrate 100 x)	1 x 120 μL	Streptavidin-HRP Diluent	1 x 12 mL
Wash Buffer (concentrate 25 x)	1 x 20 mL	Substrate Solution	1 x 10 mL
Stop Solution	1 x 6 mL	Adhesive Films	4

Specificity

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

Recovery

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

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 Horse CFTR and their serial dilutions. The results were demonstrated by the percentage of calculated concentration to the expected.

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.