IGFBP3 Kit ELISA

N° du produit ABIN5680713

Détail du produit IGFBP3 Kit ELISA

Antigène: IGFBP3 (Insulin-Like Growth Factor Binding Protein 3 (IGFBP3))

Épitope: intact

Reactivité: Humain

Méthode de détection: Colorimetric

Type de méthode: Sandwich ELISA

Gamme de detection: 3.5 ng/mL - 117 ng/mL

Seuil minimal de détection: 3.5 ng/mL

Application: ELISA

Fonction: The Intact IGFBP-3 enzyme linked immunosorbent assay (ELISA) kit provides materials for the quantitative measurement of intact IGFBP-3 in human serum and other biological fluids.

Type d'échantillon: Plasma, Serum

Analytical Method: Quantitative

Sensibilité: 1.37 ng/mL

Ingrédients:

• IGFBP-3 Calibrators A
• Cal-149F Intact IGFBP-3 Calibrators B - F (Lyophilized)
• Intact IGFBP-3 Controls I & II (Lyophilized)
• IGFBP-3 Coated Microtitration strips
• Sample Buffer I
• Sample Buffer II
• IGFBP-3 Assay Buffer
• Intact IGFBP-3 Antibody-Enzyme Conjugate Ready-to-Use
• TMB Chromogen Solution
• Stopping Solution
• Wash Concentrate A

Matériel non inclus:

1. Microtitration plate reader capable of absorbance measurement at 450 n, 405 nm and 630 nm.
2. Microplate orbital shaker.
3. Microplate washer.
4. Semi-automated/manual precision pipette to deliver 10-250 μL.
5. Repeater pipette.
6. Vortex mixer.
7. Deionized water.

Information d'application

Volume d'échantillon: 25 μL

Durée du test: 1.5 h

Plaque: Pre-coated

Préparation des réactifs:

1. IGFBP-3 calibrators B-F and IGFBP-3 Controls I & II: Tap and reconstitute IGFBP-3 Calibrator B-F and IGFBP-3 Controls I & II each with 0.5 mL deionized water. Solubilize the calibrators and controls for 15 mins in deionized water and mix well before use.
2. Wash Solution: Dilute wash concentrate 25-fold with deionized water. The wash solution is stable for one month at room temperature (23 ± 2 °C) when stored in a tightly sealed bottle.
3. Microtitration Wells: Select the number of coated wells required for the assay. The remaining unused wells should be placed in the resealable pouch with a desiccant. The pouch must be resealed to protect from moisture.

Prélèvement de l'échantillon:

• Serum is the recommended sample type.
• Sample handling, processing, and storage requirements depend on the brand of blood collection tube that you use. Please reference the manufacturer's instructions for guidance. Each laboratory should determine the acceptability of its own blood collection tubes and serum separation products.
• Samples may be stored at 4 °C if assayed within 24 hours, otherwise samples must be stored at -20 °C or below to avoid loss of bioactivity and contamination.
• Avoid assaying lipemic, hemolyzed or icteric samples.
• Avoid repeated freezing and thawing of samples. Thaw samples no more than 3 times.
• For shipping, place specimens in leak proof containers in biohazard specimen bags with appropriate specimen identification and test requisition information in the outside pocket of the biohazard specimen bag. Follow DOT and IATA requirements when shipping specimens.

Procédure de l'essai:

Allow all specimens and reagents to reach room temperature (23 ± 2 °C) and mix thoroughly by gentle inversion before use. Calibrators, controls, and unknowns should be assayed in duplicate. Note: Any sample reading higher than the highest Calibrator should be appropriately diluted with the 0 ng/mL (CAL A) and re-assayed.

1. Label the microtitration strips to be used.
2. Pipette 25 μL of the Calibrator and Controls and treated Unknowns (see sample preparation section) to the appropriate wells.
3. Add 100 μL of the IGFBP-3 Assay Buffer to each well using a repeater pipette.
4. Incubate the plate, shaking at a fast speed (600-800 rpm) on an orbital microplate shaker, for 60 minutes at room temperature (23 ± 2 °C).
5. Aspirate and wash each strip 5 times (350 μL/per well) with Wash Solution using an automatic microplate washer.
6. Add 100 μL of the IGFBP-3 Enzyme Conjugate Solution (RTU) to each well using a repeater pipette.
7. Incubate the plate, shaking at a fast speed (600-800 rpm) on an orbital microplate shaker, for 30 minutes at room temperature (23 ± 2 °C).
8. Aspirate and wash each strip 5 times with the Wash Solution (350 μL/per well) using an automatic microplate washer.
9. Add 100 μL of the TMB chromogen solution to each well using a precision pipette. Avoid exposure to direct sunlight.
10. Incubate the wells, shaking at 600-800 rpm on an orbital microplate shaker, for 8-12 min at room temperature (23 ± 2 °C). NOTE: Visually monitor the color development to optimize the incubation time.
11. Add 100 μL of the stopping solution to each well using a precision pipette. Read the absorbance of the solution in the wells within 20 minutes, using a microplate reader set to 450 nm. NOTE: Zero calibrator should be programmed as ""Blank"" while reading the optical density. If instrument has a wavelength correction, set the instrument to dual wavelength measurement at 450 nm with background wavelength correction at 630 nm.

Calcul des résultats:

NOTE The results in this package insert were calculated by plotting the log optical density (OD) data on the y-axis and log IGFBP-3 concentration on X-axis using a linear regression curve-fit.

1. Optimum results can be obtained at incubation temperature of (23 ± 2 °C).
2. Calculate the mean optical density (OD) for each Calibrator, Control, or Unknown.
3. Plot the log of the mean OD readings for each of the Calibrators along the y-axis versus log of the Intact IGFBP-3 concentrations in ng/mL along the x-axis, using a Liner regression curve-fit.
4. Determine the Intact IGFBP-3 concentrations of the Controls and unknowns from the calibration curve by matching their mean OD readings with the corresponding Intact IGFBP-3 concentrations.
5. Any sample reading higher than the highest Calibrator should be appropriately diluted with the 0 ng/mL (CAL A ) and re-assayed.
6. Any sample reading lower than the analytical sensitivity should be reported as such.
7. Multiply the measured concentrations in ng/mL by the dilution factor (20X).

Restrictions: For Research Use only

Stockage

Précaution d'utilisation: The following precautions should be observed: a) Follow good laboratory practice. b) Use personal protective equipment. Wear lab coats and disposable gloves when handling immunoassay materials. c) Handle and dispose of all reagents and material in compliance with applicable regulations WARNING: Potential Biohazardous Material This reagent may contain some human source material (e.g. serum) or materials used in conjunction with human source materials. Handle all reagents and patient samples at a Biosafety Level 2, as recommended for any potentially infectious human material in the Centers for Disease Control/National Institutes of Health manual "Biosafety in Microbiological and Biomedical Laboratories," 5th Edition, 2007.11 WARNING: Potential Chemical Hazard Some reagents in this kit contain Pro-Clean 400 and Sodium azide12 as a preservative. Pro-Clean 400 and Sodium azide in concentrated amounts are irritants to skin and mucous membranes. For further information regarding hazardous substances in the kit, please refer to the MSDS.

Stock: 4 °C

Détail du antigène

Antigène: IGFBP3 (Insulin-Like Growth Factor Binding Protein 3 (IGFBP3))

Autre désignation: IGFBP-3 (IGFBP3 Produits)

Synonymes: BP-53 Kit ELISA, IBP3 Kit ELISA, AI649005 Kit ELISA, IGFBP-3 Kit ELISA, IGgfbp3 Kit ELISA, IGF-BP3 Kit ELISA, zgc:91788 Kit ELISA, IGFBP3 Kit ELISA, igfbp3 Kit ELISA, insulin like growth factor binding protein 3 Kit ELISA, insulin-like growth factor binding protein 3 Kit ELISA, IGFBP3 Kit ELISA, Igfbp3 Kit ELISA, igfbp3 Kit ELISA, IGFBP-3 Kit ELISA, LOC100305016 Kit ELISA

Pathways: Myometrial Relaxation and Contraction, Regulation of Muscle Cell Differentiation, Skeletal Muscle Fiber Development, Regulation of Carbohydrate Metabolic Process, Autophagy, Smooth Muscle Cell Migration, Growth Factor Binding