Growth hormone (GH) or somatotropin, also known as human growth hormone (hGH or HGH) in its human form, is a peptide hormone that stimulates growth, cell reproduction, and cell regeneration in humans and other animals. It is thus important in human development. GH also stimulates production of IGF-1 and increases the concentration of glucose and free fatty acids
Reagent preparation
Prepare only the appropriate amount of required reagent on the day of use. Store all reagents as per instructions stated on the label.
1. Wash Buffer (1X) Preparation: Dilute wash buffer concentrate with ultra-pure water 1/20 before use (for example add 25mL concentrate to 475mL ultrapure water). Mix well.
2. Secondary Antibody (1X) Preparation: Dilute secondary antibody with assay buffer 1/1000 before use (for example add 12ul to 12mL of assay buffer). Mix well.
3. Detection Reagent (1X) Preparation: Dilute detection reagent with assay buffer 1/1000 before use (for example add 12ul concentrate to 12ml of assay buffer). Mix well.
4. Calibrator Preparation: Dilute the calibrator from 1mg/ml down to 5ug/ml by pipetting 5uL of calibrator stock into 995uL assay buffer. Label "Cal. Int." Mix well. Prepare calibrators with concentrations ranging from 500 ng/ml to 9.375 ng/ml. The following is an example calibrator curve.
Results calculation
1. Construct a standard curve by plotting the absorbance obtained from each standard against concentration. Use a 4 or 5 parameter curve fit.
2. The concentration of the unknowns can be back calculated directly from this standard curve using the absorbance value for each sample.
3. Any sample diluted more or less than the standard series will need additional data correction. For example, if the sample is diluted 1/50, then the concentration will be calculated by dividing by 2 due to the calibrators being 2 times more diluted. Similarly, if the sample is diluted 1/500, then the concentration will be calculated by multiplying by a correction factor of 5 due to the calibrators being 5 times more concentrated.
Sample preparation
Dilute calibrators and test samples 1/50 with assay buffer (for example add 5uL of prepared calibrator or sample to 245uL of assay buffer). Mix well. Do not store diluted samples.
< 25% for ULOQ and LLOQ and <20% for the remaining concentrations
Notes
This kit is compatible with EDTA-plasma, heparinplasma and serum samples. Samples can be stored at or below -20°C for up to 1 year.
Usage Notes
Quantification of human growth hormone in biological matrices
Protocol
This assay employs the indirect sandwich enzyme immunoassay technique. Anti-GH is coated onto a 96 well microplate. Calibrator and test samples prepared by dilution into assay buffer and are pipetted into the appropriate wells. GH present in biological matrices is bound by the immobilized anti- GH antibody. After washing away any unbound substances, tagged antiGH secondary antibody is added to the wells. After washing, diluted detection reagent is added to the wells. The plate is washed to remove any unbound antibody-enzyme reagent and a substrate solution is added to the wells for color development. The color development is proportional to the amount of GH present in test samples. The color development is stopped and the intensity of the color is measured.
Gene ID
2688
Applications
ELISA
Storage
-20C, 1 year
Reviews of Human growth hormone (hGH) (Somatropin) Pharmacokinetic ELISA