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IGFBPs (Insulin-like growth factor binding proteins) are found in various body fluids such as blood serum and amniotic fluid. They are synthesized in the liver and are produced also by various tumor cell lines and cancer cell types. IGFBP-2 is involved in neurogenesis. It is also malignancy-associated protein measurable in tumors and blood.
Elevated levels of IGFBP2 have been detected in the serum of prosate, breast, glioma, and hepatocellular carcinoma cancer patients.
Figure: IGFBP2 Standard Curve
Suggested Application:Quantitative Protein Detection; Establishing Normal Range; Validation of Antibody Array Results.
· Pre-Coated 96-Well Strip Microplate
· Wash Buffer
· Stop Solution
· Assay Diluent(s)
· Lyophilized Standard
· Biotinylated Detection Antibody
· Streptavidin-Conjugated HRP
· TMB One-Step Substrate
Other Materials Required:
· Distilled or deionized water
· Precision pipettes to deliver 2 μl to 1 ml volumes
· Adjustable 1-25 ml pipettes for reagent preparation
· 100 ml and 1 liter graduated cylinders
· Tubes to prepare standard and sample dilutions
· Absorbent paper
· Microplate reader capable of measuring absorbance at 450nm
· Log-log graph paper or computer and software for ELISA data analysis
1.Prepare all reagents, samples and standards as instructed in the manual.
2.Add 100 μl of standard or sample to each well. Incubate 2.5 h at RT or O/N at 4°C.
3.Add 100 μl of prepared biotin antibody to each well. Incubate 1 h at RT.
4.Add 100 μl of prepared Streptavidin solution to each well. Incubate 45 min at RT.
5.Add 100 μl of TMB One-Step Substrate Reagent to each well. Incubate 30 min at RT.
6.Add 50 μl of Stop Solution to each well. Read at 450 nm immediately.
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