Haptoglobin (HP) ELISA Kit

Antigen: Haptoglobin (HP)

Synonyme: HP, DKFZp470F1933, LOC100008816, HPR, MGC133752, HP-1, fb64e01, wu:fb64e01, BP, HPA1S, HP2ALPHA2, MGC111141, Ba1-647

Reaktivität: Ziege

Applikation: ELISA

Zertifikate: ISO 9001:2008

Produktnummer: ABIN456929

Produktbeschreibung

Weitere Bezeichnung: Haptoglobin (Hpt/HP)

Proben: Serum, Plasma

Spezifität: This assay recognizes goat hpt. No significant cross-reactivity or interference was observed.

Sensitivität: The minimum detectable dose of goat hpt is typically less than 0.5 ng/ml. The sensitivity of this assay, or Lower Limit of Detection (LLD) was defined as the lowest protein concentration that could be differentiated from zero. Detection Range: The minimum detectable dose of goat hpt is typically less than 0.5 ng/ml. The sensitivity of this assay, or Lower Limit of Detection (LLD) was defined as the lowest protein concentration that could be differentiated from zero.

Anwendungen

Prinzip: The microtiter plate provided in this kit has been pre-coated with goat hpt. Standards or samples are then added to the appropriate microtiter plate wells with Horseradish Peroxidase (HRP) -conjugated antibody preparation specific for goat hpt ， mix well and incubated. The more the amount of goat hpt in samples, the less HRP-conjugated antibody preparation specific for goat hpt bound by pre-coated goat hpt. Then a TMB (3,3',5,5' tetramethyl-benzidine) substrate solution is added to each well. And the color develops in opposite to the amount of goat hpt in the sample. The color development is stopped and the intensity of the color is measured.

Protokoll: Reagent Preparation:
Bring all reagents to room temperature before use. 1. Wash Buffer If crystals have formed in the concentrate, warm up to room temperature and mix gently until the crystals have completely dissolved. Dilute 20 ml of Wash Buffer Concentrate into deionized or distilled water to prepare 500 ml of Wash Buffer. 5.
Assay Procedure:
Bring all reagents and samples to room temperature before use. It is recommended that all samples, standards, and controls be assayed in duplicate. All the reagents should be added directly to the liquid level in the well. The pipette should avoid contacting the inner wall of the well. 1. Add 50μl Standard or Sample to per well. Add 50 μ l HRP- conjugate to each well immediately. Mix well with the pipette or shake the plate gently for 60 seconds. 2. Then incubate for 40 minutes at 37° C. 3. Aspirate each well and wash, repeating the process five times for a total of five washes. Wash: Fill each well with Wash Buffer (200μl) and let it stand for 2 minutes, then remove the liquid by flicking the plate over a sink. The remaining drops are removed by patting the plate on a paper towel. Complete removal of liquid at each step is essential to good performance. 7 4. Add 90 μl of TMB Substrate to each well. Incubate for 20 minutes at 37°C. Keeping the plate away from drafts and other temperature fluctuations in the dark. 5. Add 50 μl of Stop Solution to each well when the last four wells containing the lowest concentration of standards develop obvious blue color. If color change does not appear uniform, gently tap the plate to ensure thorough mixing. 6. Determine the optical density of each well within 15 minutes, using a microplate reader set to 450 nm.
Calculation of Results:
Using the professional soft ",Curve Exert 1.3", to make a standard curve is recommended, which can be downloaded from our web. Average the duplicate readings for each standard, control, and sample and divide the average zero standard optical density. Create a standard curve by reducing the data using computer software. As an alternative, construct a standard curve by plotting the absorbance ratio 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 goat hpt concentrations versus the ratio and the best fit line can be determined by regression analysis. This procedure 8 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 with the appropriate Diluent 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 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.
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. Centrifuge the sample again after thawing before the assay. Avoid repeated freeze-thaw cycles. Recommend to dilute the serum or plasma samples with Sample Diluent(1:500) before test. The suggested 500-fold dilution can be achieved by adding 10μl sample to 190μl of 6 Sample Diluent first, then complete the 500-fold dilution by adding 10μl of this solution to 240μl of Sample Diluent. The recommended dilution factor is for reference only. The optimal dilution factor should be determined by users according to their particular experiments. Note: Grossly hemolyzed samples are not suitable for use in this assay.

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

Applikationshinweise: Detection Wavelength: 450 nm

Bestandteile: Reagent (Quantity): Assay plate (1), Standard 8x0.5ml Sample Diluent (2x20ml), HRP- Conjugate (1x6ml), Wash Buffer (25×concentrate) (1x20ml), TMB Substrate (1x10ml), Stop Solution (1x10ml)

Benötigtes Material: Microplate reader capable of measuring absorbance at 450 nm, with the correction wavelength set at 540 nm or 570 nm. Pipettes and pipette tips. Deionized or distilled water. Squirt bottle, manifold dispenser, or automated microplate washer. An incubator which can provide stable incubation conditions up to 37°C±0.5°C.

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, provided it is stored as prescribed above. Refer to the package label for the expiration date. 2. Opened test plate should be stored at 2-8 C in the aluminum foil bag with desiccants to minimize exposure to damp air. The 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.

Beschränkungen: Nur für Forschungszwecke einsetzbar