Kininogen Kit ELISA

N° du produit ABIN956045

Ce kit ELISA Colorimetric est destiné à la mesure quantitative de Rat Kininogen.

Aperçu rapide pour Kininogen Kit ELISA (ABIN956045)

Antigène: Kininogen (KNG)

Reactivité: Rat

Méthode de détection: Colorimetric

Type de méthode: Sandwich ELISA

Application: ELISA

Détail du produit Kininogen Kit ELISA

Fonction: The Rat Thiostatin ELISA is a highly sensitive two-site enzyme linked immunoassay (ELISA) for the quantitative determination of thiostatin in rat biological samples.

Analytical Method: Quantitative

Attributs du produit: Thiostatin is a major protease inhibitor in serum and an acute phase protein that increases significantly in concentration in the rat as a result of inflammation. The major pathophysiological role for rat thiostatin has yet to be conclusively defined.In this assay the thiostatin present in samples reacts with the anti-thiostatin antibodies, which have been adsorbed to the surface of polystyrene microtiter wells. After the removal of unbound proteins by washing, anti-Thiostatin antibodies conjugated with horseradish peroxidase (HRP), are added. This HRP-conjugated antibody forms a complex with the previously bound thiostatin. Following another washing step, the enzyme bound to the immunosorbent is assayed by the addition of a chromogenic substrate, 3,3',5,5'-tetramethylbenzidine (TMB). The quantity of bound enzyme varies directly with the concentration of thiostatin in the sample tested, thus, the absorbance, at 450 nm, is a measure of the concentration of thiostatin in the test sample. The quantity of thiostatin in the test sample can be interpolated from the calibration curve constructed from the calibrators, and corrected for serum dilution.

Ingrédients: 1. Diluent Concentrate: One bottle containing 50 mL of a 5X concentrated diluent running buffer.
2. Wash Solution Concentrate: One bottle containing 50 mL of a 20X concentrated wash solution.
3. Enzyme-Antibody Conjugate Concentrate: One vial containing 150 µL of a 100X concentrated affinity-purified anti-Rat thiostatin antibody conjugated with HRP in a stabilizing buffer.
4. TMB Substrate Solution: One vial containing 12 mL of TMB and hydrogen peroxide in citric acid buffer at pH 3.3. WARNING: Avoid contact with skin.
5. Stop Solution: One vial containing 12 mL of 0.3 M sulfuric acid. WARNING: Avoid contact with skin.
6. Microtiter Plate: Twelve removable eight-well strips in well holder frame. Wells are coated with affinity-purified anti-rat thiostatin.
7. Rat Thiostatin Calibrator: One vial containing Rat Thiostatin Calibrator.

Matériel non inclus: Test tubes
Precision pipettes (2 µL to 200 µL)
Microplate washer/aspirator
Distilled or de-ionized H2O Microplate reader
Assorted glassware for the preparation of reagents and buffer solutions
Timer
Vortex mixer

Information d'application

Plaque: Pre-coated

Préparation des réactifs:

1. Diluent Concentrate: The Diluent Solution supplied is a 5X concentrate and must be diluted 1:5 with distilled or de-ionized water.
   2. Wash Solution Concentrate: The Wash Solution supplied is a 20X concentrate and must be diluted 1:20 with distilled or de-ionized water. Crystal formation in the concentrate is not uncommon when storage temperatures are low. Warming of the concentrate to 30-35°C before dilution can dissolve crystals.
   3. Enzyme-Antibody Conjugate Concentrate: Calculate the required amount of working conjugate solution for each microtiter plate test strip by adding 10 µL Enzyme-Antibody Conjugate to 990 µL of 1X Diluent for each test strip to be used for testing. Mix uniformly, but gently. Avoid foaming.
   4. TMB Substrate Solution: Ready to use as supplied.
   5. Stop Solution: Ready to use as supplied.
   6. Microtiter Plate: Ready to use as supplied. Unseal Microtiter Pouch and remove plate from pouch. Remove all strips and wells that will not be used in the assay and place back in pouch and re-seal along with desiccant.
   7. Rat Thiostatin Calibrator: The calibrator is now at a concentration of 800 ng/mL. The Rat Thiostatin calibrators need to be prepared immediately prior to use. Mix well between each step. Avoid foaming.

Calcul des résultats:

1. Subtract the average background value from the test values for each sample.
   2. Using the results observed for the calibrators construct a calibration curve. The appropriate curve fit is that of a four- parameter logistics curve. A second order polynomial (quadratic) or other curve fits may also be used.
   3. Interpolate test sample values from the calibration curve. Correct for serum dilution factor to arrive at thiostatin concentration in original sample.

Restrictions: For Research Use only

Stockage

Stock: 4 °C

Stockage commentaire: 1. Complete Kit: The expiration date for the kit is stated on the outer label. The recommended storage temperature is 4°C. Note: See long-term storage recommendations below for the Rat Thiostatin Calibrator.
2. Diluent: Concentrate The 5X Diluent Concentrate is stable until the expiration date. The 1X working solution is stable for at least one week from the date of preparation. Both solutions should be stored at 4°C.
3. Wash Solution: Concentrate The 20X Wash Solution Concentrate is stable until the expiration date. The 1X working solution is stable for at least one week from the date of preparation. Both solutions can be stored at room temperature (RT, 16-25°C) or at 4°C.
4. Enzyme-Antibody Conjugate: Concentrate Undiluted anti-Thiostatin-HRP conjugate should be stored at 4°C and diluted immediately prior to use. The working conjugate solution is stable for up to 8 hours.
5. TMB Substrate Solution: The TMB Substrate Solution should be stored at 4°C and is stable until the expiration date.
6. Stop Solution: The Stop Solution should be stored at 4°C and is stable until the expiration date.
7. Microtiter Plate: Anti-Rat Thiostatin coated wells are stable until the expiration date and should be stored at 4°C in the sealed foil pouch with a desiccant pack.
8. Rat Thiostatin Calibrator: Aliquot the Rat Thiostatin Calibrator and store frozen. Long Term Storage : Upon receipt, aliquot the calibrator and store them frozen. They will be stable until expiration date.
Short Term Storage : Upon receipt, aliquot the calibrator and store them frozen. They will be stable until expiration date. Short Term. Storage : Upon receipt, aliquot the calibrator and store them frozen. They will be stable until expiration date. Short Term

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

Détail du antigène

Antigène: Kininogen (KNG)

Autre désignation: Thiostatin