NEW DetectX® P
Total Cyclic AMP EIA KitsCatalog Numbers:
One plate K019-H1 Offer Price 200.00 UK Pounds
Five plate K019-H5 Offer Price 820.00 UK Pounds
Cells to Results in < 3 Hours
Features
Colour-coded, simple protocol
Reduced sample requirements
Measure to less than 4 fmol cAMP per sampleHigh optical density in 2.5 hours
INTRODUCTION
The DetectX® Cyclic AMP (cAMP) Immunoassay kit is designed to quantitatively measure cAMP present in lysed cells, tissue, plasma, urine, saliva and tissue culture media samples. The kit is unique in that all samples and standards are diluted into an acidic Sample Diluent which contains special additives and stabilizers designed to lyse cells for measurement. AcAMP standard is provided to generate a standard curve for the assay. Aspecial Plate Primer is added to all the wells and standards or samples in Sample Diluent are pipetted into the primed clear microtiter plate. AcAMP-peroxidase conjugate is added to the standards and samples in the wells. The binding reaction is initiated by the addition of an antibody to cAMP. After 2 hours, the plate is washed and substrate is added. The substrate reacts with the bound cAMP-peroxidase conjugate. After a short incubation, the reaction is stopped and the intensity of the generated color is detected in a microtiter plate reader capable of measuring 450nm wavelength. The concentration of cAMP in the sample is calculated, after making suitable correction for the dilutions. An optional Acetylation Format allows super low concentrations of cAMP to be measured.
Sample PreparationAll samples are diluted in the Sample Diluent. Tissue, urine and saliva samples should be run in the Regular Format. Plasma and samples with low expected concentrations of cAMPshould be run in the Acetylated Format. TCM samples should be diluted in TCM and read off a standard curve generated in the same TCM.
Assay Protocol
• ipet Plate Primer into all wells used for the assay.
• Add 50 μL of samples or standards into duplicate wells in the microtiter plate.
• Add 25 μL of the DetectX® cAMP-Peroxidase Conjugate, followed by 25 μL of cAMP Antibody and shake at room temperature for 2 hours.
• Wash 4 times with wash buffer.
• Add 100 μL of TMB substrate to each well and incubate for 30 minutes.
• Add 50 μL of stop solution to each well and read at 450 nm.
Sensitivity
Sensitivity was 3.9 fmol/well in the Acetylated Format.