Fetuin Kit ELISA

N° du produit ABIN956179

Détail du produit Fetuin Kit ELISA

Antigène: Fetuin

Reactivité: Humain

Méthode de détection: Colorimetric

Type de méthode: Sandwich ELISA

Application: ELISA

Fonction: The Human Fetuin ELISA is a highly sensitive two-site enzyme-linked immunoassay (ELISA) for the quantitative determination of fetuin in human serum or plasma.

Analytical Method: Quantitative

Attributs du produit: In this assay the Fetuin present in samples reacts with the anti-Fetuin antibodies which have been adsorbed to the surface of polystyrene microtiter wells. After the removal of unbound proteins by washing, anti-Fetuin antibodies conjugated with horseradish peroxidase (HRP), are added. These enzyme-labeled antibodies form complexes with the previously bound Fetuin. 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 Fetuin in the sample tested, thus, the absorbance, at 450 nm, is a measure of the concentration of Fetuin in the test sample. The quantity of Fetuin in the test sample can be interpolated from the calibration curve constructed from the calibrators, and corrected for sample 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-human Fetuin antibody conjugated with HRP in stabilizing buffer.
4. TMB Substrate Solution: One vial containing 12 mL of TMB and hydrogen peroxide in citric acid buffer at pH 3.3.
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-human Fetuin.
7. Human Fetuin Calibrator: One vial containing a lyophilized Human Fetuin Calibrator.

Matériel non inclus: Precision pipettes (2 µL to 200 µL) for making and dispensing dilutions
Test tubes
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. Human Fetuin Calibrator: Add 1.0 mL of distilled or de-ionized water to the lyophilized Human Fetuin Calibrator and mix gently until dissolved. The calibrator is now at a concentration of 7 µg/mL (the reconstituted calibrator should be aliquoted and frozen if future use is intended). Prepare the Human Fetuin Calibrators immediately prior to use according to the table below. 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, although a second order polynomial (quadratic) or other curve fits may also be used.
   3. Interpolate test sample values from calibration curve. Correct for sample dilution factor to arrive at Fetuin concentration in original sample.
   
   1. Reliable and reproducible results will be obtained when the assay procedure is carried out with a complete understanding of the information contained in the package insert instructions and with adherence to good laboratory practice.
   2. Factors that might affect the performance of the assay include proper instrument function, cleanliness of glassware, quality of distilled or de-ionized water, and accuracy of reagent and sample pipettings, washing technique, incubation time or temperature.

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 Human Fetuin Calibrator.
2. Diluent: 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: 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: Undiluted anti-Fetuin-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-human Fetuin coated wells are stable until the expiration date, and should be stored at 4°C in the sealed foil pouch with desiccant pack.
8. Human Fetuin Calibrator: The lyophilized Human Fetuin Calibrator should be stored at 4°C or frozen until reconstituted. The reconstituted calibrator should be aliquoted and stored frozen. Avoid multiple freeze/thaw cycles. The working calibrator solutions should be prepared immediately prior to use and are stable for up to 8 hours.

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

Détail du antigène

Antigène: Fetuin

Autre désignation: Fetuin (Alpha 2 HS Glycoprotein) (Fetuin Produits)

Synonymes: AHSG Kit ELISA, FETUA Kit ELISA, alpha 2-HS glycoprotein Kit ELISA, AHSG Kit ELISA

Sujet: Human Fetuin (Alpha 2 HS Glycoprotein) is a plasma protein synthesized in the liver and enriched in bone. The concentration of fetuin changes in various physiological conditions and is highest in bone during growth. This protein is a negative acute-phase protein whereas its level declines following infection, inflammation and malignancy.