IGF1 ELISA Kit

Produktnummer ABIN3073439

Dieses Colorimetric ELISA-Kit wurde entwickelt für die quantitative Messung von Maus und Ratte IGF1.

Kurzübersicht für IGF1 ELISA Kit (ABIN3073439)

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

Bindungsspezifität: Free Chain

Reaktivität: Maus, Ratte

Nachweismethode: Colorimetric

Methodentyp: Sandwich ELISA

Detektionsbereich: 0.25-10.03 ng/mL

Applikation: ELISA

Proben: Plasma, Serum

Produktdetails IGF1 ELISA Kit

Untere Nachweisgrenze: 0.25 ng/mL

Verwendungszweck: The Free 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.

Analytische Methode: Quantitative

Sensitivität: 0.1108 ng/mL

Bestandteile:

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

Benötigtes Material:

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.

Anwendungsinformationen

Probenmenge: 10 μL

Testdauer: 1 h

Plattentyp: Pre-coated

Aufbereitung der Reagenzien:

1. Total Inhibin Calibrators A-F: Tap and reconstitute Total Inhibin Calibrators A-F with 1.0 mL deionized water. Solubilize for 10 minutes, mix well and use after reconstitution.
2. 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.
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.
4. Total Inhibin Antibody-Biotin Conjugate Solution: The Total Inhibin Antibody-Biotin Conjugate Concentrate should be diluted at a ratio of 1 part conjugate to 50 parts of Inhibin B Conjugate Diluent, according to the number of wells used. If an entire plate is to be used pipet exactly 220 μL of the Concentrate in to 11 mL of the diluent.

Probennahme:

• 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 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

Testdurchführung:

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. Pipette 10 μL each of the Calibrator, Controls and Unknowns to the appropriate wells.
3. Add 100 μL of the IGF-I Enzyme Conjugate-RTU 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 with Wash Solution (350μL/per well) using an automatic microplate washer.
6. Add 100 μL of the TMB chromogen solution to each well using a repeater pipette. Avoid exposure to direct sunlight.
7. 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.
8. 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.

Ergebnisberechnung:

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.

Beschränkungen: Nur für Forschungszwecke einsetzbar

Handhabung

Vorsichtsmaßnahmen: 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, 200 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.

Lagerung: 4 °C

Details zu IGF1

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

Andere Bezeichnung: IGF-I

Pathways: RTK Signalweg, 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, Autophagie, Smooth Muscle Cell Migration, Activated T Cell Proliferation, Positive Regulation of fat Cell Differentiation