Retinol Binding Protein 4, Plasma (RBP4) ELISA Kit

Antigen: Retinol Binding Protein 4, Plasma (RBP4)

Synonyme: rbp4, MGC54038, RBP4, MGC107934, RBP, PRBP

Reaktivität: Maus

Applikation: ELISA

Zertifikate: ISO 9001:2008

Produktnummer: ABIN367944

Produktbeschreibung

Produktmerkmale: This immunoassay kit allows for the in vitro quantitative determination of mouse RBP-4 concentrations in urine, serum, plasma and other biological fluids.

Weitere Bezeichnung: Retinol-binding protein 4 (RBP-4)

Beschreibung: RBP4 is a specific carrier for retinol in the blood. It is a member of the lipocalin protein family. Under physiological conditions, RBP binds transthyretin (TTR) to prevent glomerular filtration of low molecular weight RBP in the kidneys. The RBP/TTR molar ration provides an indirect way to indicate marginal vitamin A deficiency. RBP4 may also play a role in insulin resistance in diabetes.

Spezifität: This assay recognizes recombinant and natural mouse RBP-4. No significant cross-reactivity or interference was observed. 3

Anwendungen

Prinzip: This assay employs the competitive inhibition enzyme immunoassay technique. An antibody specific to RBP-4 has been pre-coated onto a microplate. Standards or samples are added to the appropriate microtiter plate wells with HRP-conjugated RBP-4 and incubated. A competitive inhibition reaction is launched between RBP-4 (Standards or samples) and HRP-conjugated RBP-4 with the pre-coated antibody specific for RBP-4. The more amount of RBP-4 in samples, the less antibody bound by HRP-conjugated RBP-4. Then the substrate solutions are added to the wells, respectively. And the color develops in opposite to the amount of RBP-4 in the sample. The color development is stopped and the intensity of the color is measured.

Protokoll: Preparation of Reagents: 1. Bring all reagents and plate to room temperature for at least 30 minutes before use. Unused wells need store at 2-8℃and avoid sunlight. 2. Wash Buffer If crystals have formed in the concentrate, warm up to room temperature and mix gently until the crystals have completely dissolved. Dilute 15 ml of Wash Buffer Concentrate into deionized or distilled water to prepare 300 ml of Wash Buffer. Precaution: The Stop Solution provided with this kit is an acid solution. Wear eye, hand, face, and clothing protection when using this material. 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 g. Remove serum and assay immediately or aliquot and store samples at -20° C. Avoid repeated freeze-thaw cycles. 5 Plasma Collect plasma using citrate, EDTA, or heparin as an anticoagulant. Centrifuge for 15 minutes at 1000 g within 30 minutes of collection. Assay immediately or aliquot and store samples at -20° C. Avoid repeated freeze-thaw cycles. Assay Procedure: Calculation of Results: 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 RBP-4 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. 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..

Applikationshinweise: When mixing or reconstituting protein solutions, always avoid foaming. To avoid cross-contamination, change pipette tips between additions of each standard level, between sample additions, and between reagent additions. Also, use separate reservoirs for each reagent. 7 When using an automated plate washer, adding a 30 second soak period following the addition of wash buffer, and/or rotating the plate 180 degrees between wash steps may improve assay precision. To ensure accurate results, proper adhesion of plate sealers during incubation steps is necessary. Substrate Solution should remain colorless until added to the plate. Keep Substrate Solution protected from light. Substrate Solution should change from colorless to gradations of blue. Stop Solution should be added to the plate in the same order as the Substrate Solution. The color developed in the wells will turn from blue to yellow upon addition of the Stop Solution. Wells that are green in color indicate that the Stop Solution has not mixed thoroughly with the Substrate Solution.

Bestandteile: Reagent Quantity Assay plate 1 Standard 5 x 0.5ml HRP-conjugate 1 x 6 ml Wash Buffer 1 x 15 ml (20×concentrate) Substrate A 1 x 7 ml Substrate B 1 x 7 ml Stop Solution 1 x 7 ml Standard S1 S2 S3 S4 S5 Concentration (ng/ml) 1.3 2.6 6.6 26.6 100 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. Refer to the package label for the expiration date. 4 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.

Lagerung: 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. Refer to the package label for the expiration date. 4 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.

Beschränkungen: Nur für Forschungszwecke einsetzbar