IGF1 ELISA Kit

Catalog No. ABIN612799

Product Details IGF1 ELISA Kit

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

Reactivity: Mouse

Detection Method: Colorimetric

Method Type: Sandwich ELISA

Minimum Detection Limit: 0.1 ng/mL

Application: ELISA

Purpose: The AssayMax Mouse IGF-1 ELISA kit is designed for detection of mouse IGF-1 in plasma, serum and cell culture supernatants

Brand: AssayMax

Sample Type: Plasma, Cell Culture Supernatant

Analytical Method: Quantitative

Components: Mouse IGF-1 Microplate: A 96 well polystyrene microplate (12 strips of 8 wells) coated with a polyclonal antibody against mouse IGF-1. Sealing Tapes: Each kit contains 3 pre-cut, pressure-sensitive sealing tapes, which can be cut to fit the format of the individual assay. Mouse IGF-1 Standard: Recombinant mouse IGF-1 in a buffered protein base (10 ng, lyophilized). 1 Biotinylated IGF-1 Antibody (100x): A 100-fold concentrated biotinylated polyclonal antibody against IGF-1 (80µl). Streptavidin-Peroxidase Conjugate (SP Conjugate): A 100-fold concentrate (90µl). MIx Diluent Concentrate (10x): A 10-fold concentrated buffered protein base (30 ml). Wash Buffer Concentrate (20x): A 20-fold concentrated buffered surfactant (30 ml). Chromogen Substrate: A ready-to-use stabilized peroxidase chromogen substrate tetramethylbenzidine (8 ml). Stop Solution: A 0.5 N hydrochloric acid to stop the chromogen substrate reaction (12 ml).

Material not included: Microplate reader capable of measuring absorbance at 450 nm. Pipettes (1-20 µL, 20-200 µL, 200-1000µLand multiple channel). Deionized or distilled reagent grade water.

Application Details

Sample Volume: 50 μL

Assay Time: 5 h

Plate: Pre-coated

Protocol: This assay employs a quantitative sandwich enzyme immunoassay technique, which measures IGF-1 in 5 hours. A polyclonal antibody specific for mouse IGF-1 has been pre-coated onto a microplate. Mouse IGF-1 in standards and samples is sandwiched by the immobilized antibody and biotinylated polyclonal antibody specific for mouse IGF-1, which is recognized by a streptavidin- peroxidase conjugate. All unbound material is then washed away and a peroxidase enzyme substrate is added. The color development is stopped and the intensity of the color is measured.

Reagent Preparation:

Freshly dilute all reagents and bring all reagents to room temperature before use. If crystals have formed in the concentrate, mix gently until the crystals have completely dissolved. MIx Diluent Concentrate (10x): Dilute the MIx Diluent Concentrate 1:10 with reagent grade water. Store for up to 1 month at 2-8°C. IGF-1 Standard: Reconstitute the 10 ng of mouse IGF-1 Standard with 1 ml of MIx Diluent to generate a stock solution of 10 ng/ml. Allow the standard to sit for 10 minutes with gentle agitation prior to making dilutions. Prepare triplicate standard points by serially diluting the Standard solution (10 ng/ml) 1:2 with equal volume of MIx Diluent to produce 5, 2.5, 1.25, 0.63, 0.32 and 0.16 ng/ml. MIx Diluent serves as the zero standard (0 ng/ml). Any remaining solution should be frozen at -20°C. Standard Point Dilution [mIGF-1] (ng/ml) P1 1 part Standard (10 ng/ml) 10.00 P2 1 part P1 + 1 part MIx Diluent 5.00 P3 1 part P2 + 1 part MIx Diluent 2.50 P4 1 part P3 + 1 part MIx Diluent 1.25 P5 1 part P4 + 1 part MIx Diluent 0.63 P6 1 part P5 + 1 part MIx Diluent 0.32 P7 1 part P6 + 1 part MIx Diluent 0.16 P8 MIx Diluent 0.00 Biotinylated IGF-1 Antibody (100x): Spin down the antibody briefly and dilute the desired amount of the antibody 1:100 with MIx Diluent. Any remaining solution should be frozen at -20°C. Wash Buffer Concentrate (20x): Dilute the Wash Buffer Concentrate 1: 20 with reagent grade water. Streptavidin-Peroxidase Conjugate (100x): Spin down the SP Conjugate briefly and dilute the desired amount of the conjugate 1:100 with MIx Diluent. Any remaining solution should be frozen at -20°C.

Sample Collection: Plasma: Collect plasma using one-tenth volume of 0.1 M sodium citrate as an anticoagulant. Centrifuge samples at 2000 x g for 10 minutes and assay. Dilute samples 1:250 into MIx Diluent. Store samples at -20°C or below for up to 3 months. Avoid repeated freeze-thaw cycles. Serum: Samples should be collected into a serum separator tube. After clot formation, centrifuge samples at 2000 x g for 10 minutes. Remove serum and assay. Dilute samples 1:250 into MIx Diluent. Store serum at -20°C or below. Avoid repeated freeze-thaw cycles. Cell Culture Supernatants: Centrifuge cell culture media at 2000 x g for 10 minutes to remove debris. Collect supernatants and assay. Store samples at -20°C or below. Avoid repeated freeze- thaw cycles.

Assay Procedure:

Prepare all reagents, working standards and samples as instructed. Bring all reagents to room temperature before use. The assay is performed at room temperature (20 - 30 °C). Remove excess microplate strips from the plate frame and return them immediately to the foil pouch with desiccant inside. Reseal the pouch securely to minimize exposure to water vapor and store in a vacuum desiccator. Add 50 µL of Standard or sample per well, and cover wells and incubate for two hours. Start the timer after the last sample addition. Wash five times with 200 µL of Wash Buffer. Invert the plate and decant the contents, and blot it on absorbent paper towel to complete remove liquid at each step. Add 50 µL of Biotinylated IGF-1 Antibody to each well and incubate for two hours. Wash five times with 200 µL of Wash Buffer. Add 50 µL of Streptavidin-Peroxidase Conjugate per well and incubate for 30 minutes. Turn on the microplate reader and set up the program in advance. Wash five times with 200 µL of Wash Buffer. Add 50 µL of Chromogen Substrate per well and incubate for about 10 minutes or till the optimal blue color density develops. Gently tap plate to ensure thorough mixing and break the bubbles in the well with pipette tip. Add 50 µL of Stop Solution to each well. The color will change from blue to yellow. Read the absorbance on a microplate reader at a wavelength of 450 nm immediately.

Calculation of Results:

Calculate the mean value of the triplicate readings for each standard and sample. To generate a Standard Curve, plot the graph using the standard concentrations on the x-axis and the corresponding mean 450 nm absorbance on the y-axis. The best-fit line can be determined by regression analysis using log-log or four-parameter logistic curve-fit. Determine the unknown sample concentration from the Standard Curve and multiply the value by the dilution factor. Standard Curve The curve is provided for illustration only. A standard curve should be generated each time the assay is performed.

Assay Precision: Intra-assay and inter-assay coefficients of variation were 4.8 % and 7.2% respectively.

Restrictions: For Research Use only

Handling

Handling Advice: The kit should not be used beyond the expiration date.

Storage: 4 °C/-20 °C

Storage Comment: Store unopened kit at 2-8°C up to expiration date. Opened MIx Diluent may be stored for up to 1 month at 2-8°C. Store reconstituted reagents at <-20°C. Opened unused strip wells may return to the foil pouch with the desiccant pack, reseal along zip-seal. May be stored for up to 1 month in a vacuum desiccator.

References for IGF1 ELISA Kit (ABIN612799)

Steger, Füller, Garcia-Cuellar, Hetzner, Slany: "Insulin-like growth factor 1 is a direct HOXA9 target important for hematopoietic transformation." in: Leukemia, Vol. 29, Issue 4, pp. 901-8, (2015) (PubMed).

Liu, Chen, zhou, Liu, Tang: "Insulin-like growth factor-1 and bone morphogenetic protein-2 jointly mediate prostaglandin E2-induced adipogenic differentiation of rat tendon stem cells." in: PLoS ONE, Vol. 9, Issue 1, pp. e85469, (2014) (PubMed).

Nicolas-Francès, Arnauld, Kaminski, Ver Loren van Themaat, Clémencet, Chamouton, Athias, Grober, Gresti, Degrace, Lagrost, Latruffe, Mandard: "Disturbances in cholesterol, bile acid and glucose metabolism in peroxisomal 3-ketoacylCoA thiolase B deficient mice fed diets containing high or low fat contents." in: Biochimie, Vol. 98, pp. 86-101, (2014) (PubMed).

Target Details for IGF1

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

Alternative Name: Insulin-like Growth Factor 1 (IGF-1) (IGF1 Products)

Synonyms: IGF-I ELISA Kit, IGF1A ELISA Kit, IGFI ELISA Kit, C730016P09Rik ELISA Kit, Igf-1 ELISA Kit, Igf-I ELISA Kit, Npt2B ELISA Kit, IGF-1 ELISA Kit, IGFIA ELISA Kit, IGF-IB ELISA Kit, igf1 ELISA Kit, IGF-1L ELISA Kit, IGF-1a ELISA Kit, IGF1 ELISA Kit, igf-1 ELISA Kit, igf1-A ELISA Kit, igf1.S ELISA Kit, xigf1 ELISA Kit, insulin like growth factor 1 ELISA Kit, insulin-like growth factor 1 ELISA Kit, insulin-like growth factor I ELISA Kit, insulin like growth factor 1 L homeolog ELISA Kit, IGF1 ELISA Kit, Igf1 ELISA Kit, LOC100136741 ELISA Kit, igf1 ELISA Kit, LOC678666 ELISA Kit, igf1.L ELISA Kit, LOC104911603 ELISA Kit

Background: Insulin-like growth factor 1 (IGF-1) is a 70 amino acid polypeptide protein hormone with molecular mass of 7.65 kDa. IGF-1 is produced primarily by the liver in response to the stimulation of growth hormone. It is transported in plasma bound to different forms of IGF-1 binding proteins. It also binds to specific IGF-1 tyrosine kinase receptor and the insulin receptor. Inhibition IFG-1 receptor reduces pancreatic cancer growth and angiogenesis. IGF-I regulates cellular proliferation, differentiation, apoptosis, and amyloid precursor protein family (4 - 5). It may be important in the pathophysiological processes underlying chronic disease, including type 2 diabetes mellitus, coronary heart disease, cancer and Alzheimer's disease (6 - 8). Increased levels of IGF lead to an increased risk of cancer. IGF-I stimulates osteoblast proliferation, bone formation, and increases bone volume. It is a potent neurotrophic as well as a neuroprotective factor found in the central and the peripheral nervous systems of the brain.

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