IGF1 ELISA Kit

Catalog No. ABIN3073440

This Mouse and Rat IGF1 ELISA Kit is a Colorimetric ELISA Kit designed to quantify Mouse and Rat IGF1.

Quick Overview for IGF1 ELISA Kit (ABIN3073440)

Target: IGF1 (Insulin-Like Growth Factor 1 (IGF1))

Binding Specificity: total

Reactivity: Mouse, Rat

Detection Method: Colorimetric

Method Type: Sandwich ELISA

Detection Range: 0.50-20.07 ng/mL

Application: ELISA

Sample Type: Plasma, Serum

Product Details IGF1 ELISA Kit

Minimum Detection Limit: 0.50 ng/mL

Purpose: The Total Rat/Mouse IGF-I enzyme linked immunosorbent assay (ELISA) kit provides materials for the quantitative measurement of IGF-I in rat and mouse serum, plasma and other biological fluids.

Analytical Method: Quantitative

Sensitivity: 0.0397 ng/mL

Components:

• Total Rat/Mouse IGF-I Calibrators A thru F
• Total Rat/Mouse IGF-I Controls I & II
• Rat/Mouse IGF-I Ab Coated Microtitration strips
• Sample Buffer I
• Sample Buffer II
• IGF-I Assay Buffer
• Total Rat/Mouse IGF-I Ab Enzyme Conjugate Ready-To-Use (RTU)
• TMB Chromogen Solution
• Stopping Solution
• Wash Concentrate A

Material not included:

1. Microtitration plate reader capable of absorbance measurement at 450 nm, 405nm and 630 nm.
2. Microplate shaker.
3. Microplate washer.
4. Semi-automated/manual precision pipette to deliver 10-250 μL.
5. Vortex mixer.
6. Deionized water.
7. Disposable 12 x 75 mm culture tubes.

Application Details

Sample Volume: 10 μL

Assay Time: 1.5 h

Plate: Pre-coated

Reagent Preparation:

1. Wash Solution: Dilute wash concentrate 25-fold with deionized water. The wash solution is stable for one month at room temperature when stored in a tightly sealed bottle.
2. 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.

Sample Collection:

• Serum or plasma 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 should be stored frozen at -20 °C or lower.
• Avoid repeated freezing and thawing of samples.
• Avoid assaying lipemic, hemolyzed or icteric samples.
• 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.

Assay Procedure:

Allow all specimens and reagents to reach room temperature and mix thoroughly by gentle inversion before use. Calibrators, controls, and unknowns should be assayed in duplicate.

1. Label the microtitration strips to be used.
2. Sample Preparation (1:10 dilution): a. In a vial, pipette 10 μL of the sample into 45 μL of IGF-I Sample Buffer I, vortex and incubate for 30 minutes at room temperature. b. Add 45 μL of IGF-I Sample Buffer II to the sample vial, vortex, and incubate for 10 minutes at room temperature. The sample is now ready to be assayed.
3. Pipette 20 μL of the Calibrator, Controls and Unknowns from step 2b to the appropriate wells.
4. Add 50 μL of the IGF-I Assay Buffer to each well using a repeater pipette.
5. 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).
6. Aspirate and wash each strip 5 times with Wash Solution (350μL/per well) using an automatic microplate washer.
7. Add 100 μL of the Rat/Mouse IGF-I Ab Enzyme Conjugate-RTU to each well using a repeater pipette.
8. 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).
9. Aspirate and wash each strip 5 times with the Wash Solution (350μL/per well) using an automatic microplate washer.
10. Add 100 μL of the TMB chromogen solution to each well using a repeater pipette. Avoid exposure to direct sunlight.
11. 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.
12. Add 100 μL of the Stopping solution to each well using a repeater 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.

Calculation of Results:

NOTE: The results in this package insert were calculated by plotting the log optical density (OD) data on the y-axis and log IGF-I concentration on X-axis using a cubic regression curve-fit. Alternatively, log vs. log quadratic regression curve-fit can be used. Other data reduction methods may give slightly different results.

1. Calculate the mean optical density (OD) for each calibrator, Control, or Unknown.
2. Plot the log of the mean OD readings for each of the Calibrators along the y- axis versus log of the IGF-I concentrations in ng/mL along the x-axis, using a cubic regression curve-fit.
3. Determine the IGF-I concentrations of the Controls and unknowns from the calibration curve by matching their mean OD readings with the corresponding IGF-I concentrations.
4. Any sample reading lower than the analytical sensitivity should be used as measured concentration.
5. Multiply the measured concentrations in ng/mL by the dilution factor (10 folds).

Restrictions: For Research Use only

Handling

Precaution of Use: For Research Use Only. Not for use in diagnostic procedures. 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 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. WARNING: Potential Chemical Hazard Some reagents in this kit may contain Pro-Clean 400 and Sodium azide 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.

Storage: 4 °C

Target Details for IGF1

Target: IGF1 (Insulin-Like Growth Factor 1 (IGF1))

Alternative Name: IGF-I

Pathways: RTK Signaling, Intracellular Steroid Hormone Receptor Signaling Pathway, Peptide Hormone Metabolism, Hormone Activity, Regulation of Intracellular Steroid Hormone Receptor Signaling, Regulation of Hormone Metabolic Process, Regulation of Hormone Biosynthetic Process, Stem Cell Maintenance, Glycosaminoglycan Metabolic Process, Regulation of Carbohydrate Metabolic Process, Autophagy, Smooth Muscle Cell Migration, Activated T Cell Proliferation, Positive Regulation of fat Cell Differentiation