The IGF-1 ELISA Assay Kit is intended for the quantification of Insulin-like Growth Factor 1 (IGF-1) in serum and plasma. The Eagle Biosciences IGF-1 ELISA Assay Kit is for research use only and should not be used for diagnostic procedures.

SKU: ARG80495 Categories: ,


IGF-I ELISA Assay is For Research Use Only

Size: 1×96 wells
Sensitivity: 0.09 ng/ml
Dynamic Range: 2-50 ng/mL
Incubation Time: 1.75 hours
Sample Type: Serum, Plasma
Sample Size: 20 µl
Alternative Names: Insulin like growth factors 1, IGF1, IGF-1

Assay Background

Insulin-like growth factors (IGF) I and II play a pivotal role in regulating the proliferation, differentiation and specific functions of many cell types. IGF-I is identical with somatomedin C (Sm-C) and has a molecular weight of 7649 Daltons. Its major regulators are growth hormone (GH) and nutrition, although its production in specific tissues is affected by a multitude of tropic hormones and other peptide growth factors. In contrast to many other peptide hormones, IGFs are avidly bound to specific binding proteins (IGFBP). the seven classes of IGFBPs which are known at present either bind IGF-I and IGF-II with similar affinities or show a preference for IGF-II.

Assay Principle

This IGF-I ELISA assay employs the quantitative sandwich enzyme immunoassay technique. An antibody specific for Human IGF-I has been pre-coated onto a microtiter plate. IGF-I in samples will be release by diluted with an acidic Sample/Standard diluent buffer from IGF1BPs. Diluted standards or samples are pipetted into the wells and any IGF-I present is bound by the immobilized antibody. Then a biotin-conjugated antibody specific for IGF-I is added to each well and incubate. Following a washing to remove unbound substances, streptavidin conjugated to Horseradish Peroxidase (HRP) is added to each microplate well and incubated. After washing away any unbound antibodyenzyme reagent, a substrate solution (TMB) is added and color develops in proportion to the amount of IGF-I bound in the initial step. The color development is stopped by the addition of acid and the intensity of thecolor is measured at a wavelength of 450nm ±2nm.The concentration of IGF-I in the sample is then determined by comparing the O.D of samples to the standard curve.

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Additional Information

Assay Procedure

  1. Remove excess microplate strips from the plate frame, return them to the foil pouch containing the desiccant pack, and reseal it.
  2. Add 80μl of Biotin-conjugated Antibody into each well.
  3. Add 20 μl of standards, diluted controls, diluted samples (dilute 1:21 with Sample/Standard diluent buffer) and zero controls (Sample/Standard diluent buffer) in duplicates into appropriate wells. Incubate for 1 h at room temperature on microplate shaker (~350rpm).
  4. Aspirate each well and wash, repeating the process 4 times for a total 5 washes. Wash by filling each well with 1× Wash Buffer (300μl) using a squirt bottle, manifold dispenser, or autowasher. Complete removal of liquid at each is essential to good performance. After the last wash, remove any remaining Wash Buffer by aspirating, decanting or blotting against clean paper towels.
  5. Add 100 μl of HRP-Streptavidin solution to each well. Cover wells and incubate for 30 minutes at room temperature on microplate shaker (~350rpm).
  6. Aspirate each well and wash as step 3.
  7. Add 100μl of TMB Reagent to each well. Incubate for 15 minutes at room temperature in dark.
  8. Add 100μl of Stop Solution to each well. The color of the solution should change from blue to yellow.
  9. Read the OD with a microplate reader at 450nm immediately. (optional: read at ≥ 590 nm as reference wavelength)

Package Inserts

Please note: All documents above are for reference use only and should not be used in place of the documents included with this physical product. If digital copies are needed, please contact us.

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