emoved by a wash step. The reaction is completed by the addition of the colorimetric substrate, 3,3,5,5-tetramethylbenzidine/hydrogen peroxide (TMB/H₂O₂). The addition of TMB/H₂O₂ to each well results in a blue-green product background, and the reaction is stopped by the addition of 1 M phosphoric acid. A bright yellow color results, and absorbance of peroxidase-labeled cAMP that is bound to the cAMP-specific antibody is determined using a spectrophoto- metric plate reader at 450 nm. Since the amount of cAMP-horseradish peroxidase bound to rabbit anti-cAMP antibody is inversely proportional to cAMP in the sample (or standard), a high absorbance value corresponds to low levels of cAMP present in the sample. The EIA cAMP Kit includes cAMP standards for calculating standard curves. As an option for increased sensitivity (figure 2), standards and prepared samples can be acetylated by treatment with acetic anhydride in the presence of triethylamine.¹ Picomolar detection is possible for acetylated cAMP, whereas nonacetylated cAMP levels can generally be assayed in the nanomolar range. The EIA cAMP Kit includes all the re- agents and protocols for quantifying cAMP levels in both nonacetylated and acetylated preparations.

AC Toxin for Increased Intracellular cAMP Levels

Stratagene is offering adenylate cyclase (AC) toxin, a useful reagent for increasing intracellular cAMP levels in cultured cells. AC toxin can be used to study cAMP-dependent protein kinase activation, the key regulatory event in signal transduction pathways involving cAMP. Modulators currently available for examining the effects of cAMP on intracellular signaling include
'"/>

Source:

Page: All 1 2 3 4 5 6
Related biology technology :

1. The Best Enzyme for the Toughest PCR Challenges Improved with New Hot-Start Feature
2. Comparing Fidelity and Performance of Proofreading PCR Enzymes
3. pfuturbo DNA Polymerase: A High-Performance, High-Fidelity Enzyme Ideal for PCR Cloning
4. Greater Amplification Specificity with New Hot Start PCR Enzyme
5. Improve Amplification Specificity with Hot Start PCR Enzyme
6. Choice of RT-PCR Enzymes
7. Properties of PCR Enzymes
8. Select the Optimal Enzyme for RT-PCR
9. Enzyme Purification With the Econo-Pac Q Cartridge
10. Bio-Plex Cytokine Immunoassays and ELISA: Comparison of Two Methodologies in Testing Samples From Asthmatic and Healthy Children, Rev A
11. Principles of Curve Fitting for Multiplex Sandwich Immunoassays, Rev B