hGH ADA (human growth hormone) ELISA
Catalogue Number: EL-141-878
Size: 1X96 wells
Antigen: antibodies to hGH
Storage: -20°C, 1 year
Reactivity: Human
Detection Range: 30 ng/mL -1920 ng/mL
Detection Minimum: 30 ng/mL
Protocol: This assay employs the indirect enzyme immunoassay technique. hGH is coated onto a 96 well microplate. Calibrator and test samples are prepared by dilution into assay buffer and are pipetted into the appropriate wells. Antibodies to hGH present in biological matrices are bound to the immobilized hGH. After washing away any unbound substances, antibody-enzyme reagent and a substrate solution is added to the wells for color development. The color development is proportional to the amount of antibodies to hGH present in the calibrator and test samples. The color development is stopped and the intensity of the color is measured.
Components: Coated microtiter plate, 96 wells (12x8 strips) 1 Calibrator diluent 1.8 ml Calibrator (1.15mg/mL) 12 μl 10X wash buffer 50 ml Assay buffer 50 ml 1000X detection reagent 17 μl TMB 12 ml TMB stop solution
Method Type: Direct sandwich ELISA
Detection Method: Peroxidase / OD450
Principle: Quantification of antibodies to hGH
Plate: Strip
Sample Type: Serum Plasma
Sample Volume: 15ul
Assay Time: 2.5 hours
Specificity: anti-hGH antibodies
Assay Precision: <20%, <20%
Preservative: none
Description: 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
Gene ID: N/ap
Assay Procedure: This assay employs the indirect enzyme immunoassay technique. hGH is coated onto a 96 well microplate. Calibrator and test samples are prepared by dilution into assay buffer and are pipetted into the appropriate wells. Antibodies to hGH present in biological matrices are bound to the immobilized hGH. After washing away any unbound substances, antibody-enzyme reagent and a substrate solution is added to the wells for color development. The color development is proportional to the amount of antibodies to hGH present in the calibrator and test samples. The color development is stopped and the intensity of the color is measured
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 deionized water 1/10 before use (for example add 20mL concentrate to 180mL deionized water). Mix well. 2. Detection Reagent (1X) Preparation: Dilute detection reagent with assay buffer 1/1000 before use (for example add 11μl concentrate to 11ml of assay buffer). Mix well. 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. Alternatively a log-log curve fit may be used. 2. The concentration of the unknowns can be read directly from this standard curve using the absorbance value for each sample. 3. Any sample undiluted or diluted still reading greater than the highest standard should be diluted appropriately with assay buffer and retested. If the samples have been diluted, the concentration determined from the standard curve must be multiplied by the dilution factor.
Sample Collection: 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.
Sample Preparation: Dilute calibrators and test samples 1/100 with assay buffer (for example add 5µL of prepared calibrator or sample to 495µL of assay buffer). Mix well. Do not store diluted samples.