Human CAMP-dependent protein kinase type I-beta regulatory subunit (PRKAR1B) ELISA Kit

Cyclic AMP-dependent protein kinase A (PKA) is an essential enzyme in the signaling pathway of the second messenger cAMP. Through phosphorylation of target proteins, PKA controls many biochemical events in the cell including regulation of metabolism, ion transport, and gene transcription. PRKAR1b is composed of 2 regulatory and 2 catalytic subunits and dissociates from the regulatory subunits upon binding of cAMP. BAD is required to assemble the complex, the lack of which results in diminished mitochondria-based glucokinase activity and blunted mitochondrial respiration in response to glucose. Glucose deprivation results in dephosphorylation of BAD, and BAD-dependent cell death.

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Human CAMP-dependent protein kinase type I-beta regulatory subunit (PRKAR1B) ELISA Kit has high sensitivity and excellent specificity for detection of Human PRKAR1B. No significant cross-reactivity or interference between Human PRKAR1B and analogues was observed.

This Human CAMP-dependent protein kinase type I-beta regulatory subunit (PRKAR1B) ELISA Kit employs a two-site sandwich ELISA to quantitate PRKAR1B in samples. An antibody specific for PRKAR1B has been pre-coated onto a microplate. Standards and samples are pipetted into the wells and anyPRKAR1B present is bound by the immobilized antibody. After removing any unbound substances, a biotin-conjugated antibody specific for PRKAR1B is added to the wells. After washing, Streptavidin conjugated Horseradish Peroxidase (HRP) is added to the wells. Following a wash to remove any unbound avidin-enzyme reagent, a substrate solution is added to the wells and color develops in proportion to the amount of PRKAR1B bound in the initial step. The color development is stopped and the intensity of the color is measured.

• Do not mix components from different kit lots or use reagents beyond the kit expiration date.• Allow all reagents to warm to room temperature for at least 30 minutes before opening.• Pre-rinse the pipet tip with reagent, use fresh pipet tips for each sample, standard and reagent to avoid contamination.• Unused wells must be kept desiccated at 4 °C in the sealed bag provided.• Mix Thoroughly is very important for the result. It is recommended using low frequency oscillator or slight hand shaking every 10 minutes.• It is recommended that all samples and standards be assayed in duplicate or triplicate.