Prealbumin ELISA Kit

Antigen: Prealbumin

Synonyms: D17860, AA408768, AI787086, MGC107649, prealbumin, TTR

Reactivity: Human

Application: ELISA

Certificates: ISO 9001:2008

Catalog no.: ABIN367140

Additional Information

Characteristics: This immunoassay kit allows for the in vitro quantitative determination of human PA concentrations in serum, plasma and other biological fluids.

Alternative name: Prealbumin (PA)

Sample Type: Serum, Plasma

Specificity: This assay recognizes human PA. No significant cross-reactivity or 3 interPArence was observed.

Application Details

Principle: The microtiter plate provided in this kit has been pre-coated with an antibody specific to PA. Standards or samples are then added to the appropriate microtiter plate wells with a Horseradish Peroxidase (HRP) -conjugated antibody preparation specific for PA and incubated. Then substrate solutions are added to each well. The enzyme-substrate reaction is terminated by the addition of a sulphuric acid solution and the color change is measured spectrophotometrically at a wavelength of 450 nm ± 2 nm. The concentration of PA in the samples is then determined by comparing the O.D. of the samples to the standard curve.

Protocol: Preparation of Reagents: Sample Collection and Storage: Serum Use a serum separator tube (SST) and allow samples to clot for 30 minutes before centrifugation for 15 minutes at 1000 x g. Remove serum and assay immediately or aliquot and store samples at _ -20° C. Avoid repeated freeze-thaw cycles. Plasma Collect plasma using citrate, EDTA, or heparin as an anticoagulant. Centrifuge for 15 minutes at 1000 x g within 30 minutes of collection. Assay immediately or aliquot and store samples at _ -20° C. Avoid repeated freeze-thaw cycles. Note: Grossly hemolyzed samples are not suitable for use in this assay. Assay Procedure: Calculation of Results: 7 Average the duplicate readings for each standard, control, and sample and subtract the average zero standard optical density. Create a standard curve by reducing the data using computer software capable of generating a four parameter logistic (4-PL) curve-fit. As an alternative, construct a standard curve by plotting the mean absorbance for each standard on the y-axis against the concentration on the x-axis and draw a best fit curve through the points on the graph. The data may be linearized by plotting the log of the PA concentrations versus the log of the O.D. and the best fit line can be determined by regression analysis. This procedure will produce an adequate but less precise fit of the data. If samples have been diluted, the concentration read from the standard curve must be multiplied by the dilution factor. Limitations of the Procedure: The kit should not be used beyond the expiration date on the kit label. Do not mix or substitute reagents with those from other lots or sources. 8 If samples generate values higher than the highest standard, dilute the samples and repeat the assay. Any variation in operator, pipetting technique, washing technique, incubation time or temperature, and kit age can cause variation in binding. This assay is designed to eliminate interPArence by soluble receptors, binding proteins, and other factors present in biological samples. Until all factors have been tested in the Immunoassay, the possibility of interPArence cannot be excluded.

Assay Procedure: Assay Time: 1-3h
Sample Volume: 50-100ul

Application Notes: Detection Wavelength: 450 nm

Components: Reagent Quantity Assay plate 1 Standard 5 x 0.5 ml HRP-conjugate 1 x 6 ml Substrate A 1 x 7 ml Substrate B 1 x 7 ml Stop Solution 1 x 7 ml 4 Standard S1 S2 S3 S4 S5 Concentration (μg/ml) 40 80 200 400 800 STORAGE 1. Unopened test kits should be stored at 2-8°C upon receipt and the microtiter plate should be kept in a sealed bag. The test kit may be used throughout the expiration date of the kit. RePAr to the package label for the expiration date. 2. Opened test kits will remain stable until the expiring date shown, provided it is stored as prescribed above. 3. A microtiter plate reader with a bandwidth of 10 nm or less and an optical density range of 0-3 OD or greater at 450nm wavelength is acceptable for use in absorbance measurement.

Storage: 1. Unopened test kits should be stored at 2-8°C upon receipt and the microtiter plate should be kept in a sealed bag. The test kit may be used throughout the expiration date of the kit. RePAr to the package label for the expiration date. 2. Opened test kits will remain stable until the expiring date shown, provided it is stored as prescribed above. 3. A microtiter plate reader with a bandwidth of 10 nm or less and an optical density range of 0-3 OD or greater at 450nm wavelength is acceptable for use in absorbance measurement. Expiration date: six months from the date of manufacture.

Restrictions: For Research Use only