Prolactin (PRL) ELISA Kit

Antigen: Prolactin (PRL)

Synonyme: Prl1a1, AV290867, PRLB, Prol, PRLSD1, RNPROL, RATPRLSD1, PRL, prolactin

Reaktivität: Fisch

Applikation: ELISA

Zertifikate: ISO 9001:2008

Produktnummer: ABIN364862

Produktbeschreibung

Produktmerkmale: This immunoassay kit allows for the in vitro quantitative determination of fish PRL concentrations in serum, plasma and other biological fluids.

Proben: Serum, Plasma

Beschreibung: Prolactin (PRL) or Luteotropic hormone (LTH) is a peptide hormone primarily associated with lactation. PRL is synthesised and secreted by lactotrope cells in the adenohypophysis. It is also produced in other tissues including the breast, the decidua, parts of the central nervous system and the immune system. Pituitary prolactin secretion is regulated by neuroendocrine neurons in the hypothalamus, the most important ones being the neurosecretory tuberoinfundibulum (TIDA) neurons of the arcuate nucleus, which secrete dopamine to act on the dopamine-2 receptors of lactotrophs, causing inhibition of prolactin secretion. Thyrotropin-releasing factor has a stimulatory effect on prolactin release. Mouse prolactin is a single-chain polypeptide hormone containing 197 amino acid residues with a molecular weight of 23 kDa. In the mouse, there is a large family of paralogous genes closely related to PRL. PRL family genes reside on chromosome 13 of the mouse genome.

Spezifität: This assay recognizes fish PRL. No significant cross-reactivity or interference was observed.

Anwendungen

Prinzip: The microtiter plate provided in this kit has been pre-coated with an antibody specific to PRL. Standards or samples are then added to the appropriate microtiter plate wells with a Horseradish Peroxidase 3 (HRP)-conjugated PRL and anti-PRL-antibody and incubated. Then substrate solution A and B are added to each well. Only those wells that contain PRL, HRP-conjugated PRL and anti-PRL-antibody will exhibit a change in color. 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 PRL in the samples is then determined by comparing the O.D. of the samples to the standard curve.

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°C and avoid sunlight. 2. Wash Buffer If crystals have formed in the concentrate, warm 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. 3. Standard Reconstitute the Standards with 0.5 ml of ddH 2 O, respectively. Allow the standard to sit for a minimum of 15 minutes 6 with gentle agitation prior to use. 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. 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 7 -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 PRL 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: 9 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. It is important that the Calibrator Diluent selected for the standard curve be consistent with the samples being assayed. If samples generate values higher than the highest standard, dilute the samples with the appropriate Calibrator Diluent and repeat the assay. Any variation in Standard Diluent, operator, pipetting technique, washing technique, incubation time or temperature, and kit age can cause variation in binding. This assay is designed to eliminate interference by soluble receptors, binding proteins, and other factors present in biological samples. Until all factors have been tested in the Immunoassay, the possibility of interference cannot be excluded.

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

Applikationshinweise: Detection Wavelength: 450 nm

Bestandteile: Reagent Quantity Assay plate 1 Standard(S 1 -S 5 ) 5 Antibody 1 x 6 ml 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 S 1 S 2 S 3 S 4 S 5 Concentration (ng/ml) 0.2 1 5 20 100 STORAGE 5 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. 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: 5 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. 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.

Forschungsgebiet: Hormone

Beschränkungen: Nur für Forschungszwecke einsetzbar