Mouse/Rat GH ELISA Assay Kit

$540.00

The Eagle Biosciences Mouse/Rat GH ELISA Assay Kit is intended to be used for the measurement of Growth Hormone in mouse and rat serum and plasma samples. The Mouse/Rat GH ELISA Assay Kit is for Research Use Only.

Mouse/Rat GH ELISA Assay Kit

For Research Use Only

Size: 1×96 wells
Sensitivity: 0.04 ng/ml
Incubation Time: 3 hours
Sample Type: Serum, Plasma
Sample Size: 20 µL

Mouse/Rat GH ELISA Assay Kit is manufactured by Mediagnost

Additional Information

Assay Principle

The Eagle Biosciences Mouse/Rat GH ELISA Assay Kit is a so-called sandwich-assay. It utilizes two different specific high affinity polyclonal antibodies for this protein. The GH in the samples binds quantitatively to the immobilized antibody. In the following step, the biotinylated antibody in turn binds GH. After washing, a streptavidin-peroxidase-enzyme conjugate will be added, which will bind highly specific to the biotin of the antibody. Subsequently, the peroxidase catalyzes an enzymatic reaction resulting in a blue coloration. The intensity of the blue color depends on the GH content of the sample. The reaction is stopped by the addition of stop solution and color intensity is quantified by measuring the absorption.

Assay Procedure

  1. Before the assay procedure, bring all reagents to room temperature (20 – 25°C).
    Add 100 µL Dilution Buffer VP into the first two wells (these wells serve as blanks). Subsequently add 100 µL Standard or 100 µL of diluted Control Sera or diluted samples into wells designated for standards A-G, controls KS1 and KS2 or samples.
  2. Cover the wells with sealing tape and incubate the plate for 1 hour at room temperature (shake at 350 rpm).
  3. After incubation, aspirate the contents of the wells and wash the wells 5 times 300 µL Washing Buffer WP / well. The washing buffer should incubate for at least for 15 seconds/cycle. Tap the plate firmly against several layers of folded paper towels to remove excess washing buffer.
  4. Following the last washing step pipette 100 µL of the Antibody Conjugate AK in each well.
  5. Cover the wells with sealing tape and incubate the plate for 1 hour at room temperature (shake 350 rpm).
  6. After incubation wash the wells 5 times with Washing Buffer as described in step 3.
  7. Following the last washing step pipette 100 µL of the Enzyme Conjugate EK in each well. Cover the wells with sealing tape and incubate the plate for 0.5 hour at room temperature (shake 350 rpm).
  8. After incubation wash the wells 5 times with Washing Buffer as described in step 3.
  9. Pipette 100 µL of the TMB Substrate Solution in each well.
  10. Incubate the plate for 30 minutes in the dark at room temperature (20 – 25°C).
  11. Stop the reaction by adding 100 µL of Stopping Solution.
  12. Measure the color reaction within 30 minutes at 450 nm (reference filter ≥590 nm).

Manual

Product Manual


Publications

Citations

Popp S., Bielohuby M., Meurer S., Horngacher A., Bildingmaier M.; Abstract-Nr. P2 8-5-Analysis of different blood sample pre-treatment conditions on hormone concentrations in rats;Quelle/ Source: Abstract-CD 55. Symposium der Deutschen Gesellschaft für Endokrinologie 2012; ISSN 1862-1503