IGFBP3 ELISA 试剂盒

• 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

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.

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.

• 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.

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.

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).