Gastrokine 2 Kit ELISA

N° du produit ABIN956185

Kit ELISA Gastrokine 2 Humain, Colorimetric test pour la quantification de Humain Gastrokine 2.

Aperçu rapide pour Gastrokine 2 Kit ELISA (ABIN956185)

Antigène: Gastrokine 2 (GKN2)

Reactivité: Humain

Méthode de détection: Colorimetric

Type de méthode: Sandwich ELISA

Application: ELISA

Détail du produit Gastrokine 2 Kit ELISA

Fonction: This ELISA kit is a sandwich enzyme immunoassay for the in vitro quantitative measurement of human GKN2 in serum and plasma.

Analytical Method: Quantitative

Attributs du produit: The microtiter plate provided in this kit has been pre-coated with antibody specific to GKN2. Calibrators and samples are then added to the plate wells and GKN2, if present, will bind to the antibody pre-coated wells. In order to quantitatively determine the amount of GKN2 present in the sample a preparation of horseradish peroxidase (HRP) conjugated polyclonal antibody specific for GKN2 is added to each well to sandwich the GKN2 immobilized on the plate. The microtiter plate undergoes incubation and the wells are thoroughly washed to remove all unbound components. Next, A and B substrate solutions are added to each well. The HRP and substrate are allowed to react over a short incubation time. Only those wells that contain GKN2 and enzyme conjugated 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.

Ingrédients: Pre-coated 96-well plate 1 Calibrator 1 (0 ng/mL)
Calibrator 2 (5 ng/mL)
Calibrator 3 (10 ng/mL)
Calibrator 4 (25 ng/mL)
Calibrator 5 (50 ng/mL)
Calibrator 6 (100 ng/mL)
Enzyme Conjugate (1 x 6 mL)
Substrate A (1 x 6 mL)
Substrate B (1 x 6 mL)
Stop Solution (1 x 6 mL)
Wash Buffer (100X concentrate) (1 x 6 mL)
Lysis Buffer 1 x 10 mL.

Matériel non inclus: 1. Microplate reader with 450 nm filter.
2. Precision pipettes to deliver 1-2 mL volumes.
3. Adjustable 10-100 mL pipettes for reagent preparation.
4. 100 mL and 1 L graduated cylinders.
5. Calibrated adjustable precision pipettes with disposable tips (multi-channel is desirable for large assays).
6. 37°C incubator.
7. Absorbent paper.
8. Distilled or deionized water
9. Data analysis tools such as graphing software, or graph paper (linear, log-log, semi-log, or log-logit as desired).
10. Tubes to prepare Calibrators or sample dilutions.

Information d'application

Plaque: Pre-coated

Préparation des réactifs:

All reagents must be allowed to reach room temperature before use. Additional information for individual reagents can be found on vial labels. Note: 1. This kit contains materials with small quantities of sodium azide. Sodium azide reacts with lead and copper plumbing to form explosive metal azides. Upon disposal, flush drain with a large volume of water to prevent azide accumulation. Avoid ingestion and contact with eyes, skin, and mucous membranes. In case of contact, rinse affected area with plenty of water. Observe all federal, state, and local regulations for disposal.
2. All blood components and biological materials should be handled as potentially hazardous. Follow universal precautions as established by the Centers for Disease Control and Prevention and by the Occupational Safety and Health Administration when handling and disposing of infectious agents.

Procédure de l'essai:

1. Prepare all calibrators before starting assay procedure (see Reagent Preparation). It is recommended that all calibrators and samples be added in duplicate to the microtiter plate.
   2. First, secure the desired number of coated wells in the holder, then add 50 µL of calibrators and samples to the appropriate well of the antibody coated microtiter plate.
   3. Add 100 µL of conjugate to each well. Mix well. Complete mixing in this step is important. Cover and incubate for 1 hour at 37°C.
   4. Prepare substrate solution no more than 15 minutes before end of incubation (see Reagent Preparation).
   5. Wash the microtiter plate using one of the specified methods indicated below:
   6. Manual Washing: Remove incubation mixture by aspiration contents of the plate into a sink or proper waste container. Using a squirt bottle, fill each well completely with distilled or de-ionized water, then aspirate contents of the plate into a sink or proper waste container. Repeat this procedure four more times for a total of five washes. After final wash, invert plate, and blot dry by hitting plate onto absorbent paper or paper towels until no moisture appears. Note: Hold the sides of the plate frame firmly when washing the plate to assure that all strips remain securely in frame.
   7. Automated Washing: Aspirate all wells, then wash plate five times using distilled or de-ionized water. Always adjust your washer to aspirate as much liquid as possible and set fill volume to 350 µL/well/wash (range: 350-400 µL). After final wash, invert plate, and blot dry by hitting plate onto absorbent paper and paper towels until no moisture appears. It is recommended that the washer be set for a soaking time of 15-30° Conds or shaking time of 5 seconds between washes.
   8. Add 50 µL substrate A and B to each well. Cover and incubate for 15 minutes at 20-25°C.
   9. Add 50 µL stop solution to each well. Mix well.
   10. Read the optical density (OD) at 450 nm using a microtiter plate reader within 30 minutes.

Calcul des résultats:

1. Calculate the mean absorbance value A450 for each set of calibrators and samples.
   2. Subtract the mean value of the zero calibrator from all other values, construct the calibration curve using graph paper of statistical software.
   3. To determine the concentration present in each samples locate the OD on the y-axis and extend a horizontal line to the calibration curve. At the point of intersection draw a vertical line to the x-axis and read the corresponding concentration.
   4. The sensitivity of this assay is 1.0 ng/mL

Restrictions: For Research Use only

Stockage

Stock: 4 °C

Stockage commentaire: All reagents should be stored at 4°C upon receipt. For expiration date refer to kit label.

Date de péremption: The expiry date is stated on the label.

Détail du antigène

Antigène: Gastrokine 2 (GKN2)