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PRKACA ELISA Kit (Human)

CAT: 0247-OKCD09352-96WSize: 96 WellDry Ice: NoHazardous: No
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CAT#:0247-OKCD09352-96WSize:96 Well
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Reproducibility
Intra-assay Precision (Precision within an assay) : 3 samples with low, middle and high level Protein Kinase, cAMP Dependent Catalytic Alpha (PRKACa) were tested 20 times on one plate, respectively. Inter-assay Precision (Precision between assays) : 3 samples with low, middle and high level Protein Kinase, cAMP Dependent Catalytic Alpha (PRKACa) were tested on 3 different plates, 8 replicates in each plate. CV (%) = SD/meanX100 Intra-Assay: CV<10% Inter-Assay: CV<12%
CAS Number
7732-18-5
Gene Name
Protein kinase cAMP-activated catalytic subunit alpha
Gene Aliases
CAFD1; cAMP-dependent protein kinase catalytic subunit alpha; PKA C-alpha; PKACA; PPNAD4; protein kinase A catalytic subunit; protein kinase, cAMP-dependent, alpha catalytic subunit; protein kinase, cAMP-dependent, catalytic, alpha.
Gene ID
5566
Accession Number
NP_001291278.1
Reactivity
Homo sapiens|Human
Target
Phosphorylates a large number of substrates in the cytoplasm and the nucleus. Regulates the abundance of compartmentalized pools of its regulatory subunits through phosphorylation of PJA2 which binds and ubiquitinates these subunits, leading to their subsequent proteolysis. Phosphorylates CDC25B, ABL1, NFKB1, CLDN3, PSMC5/RPT6, PJA2, RYR2, RORA and VASP. RORA is activated by phosphorylation. Required for glucose-mediated adipogenic differentiation increase and osteogenic differentiation inhibition from osteoblasts. Involved in the regulation of platelets in response to thrombin and collagen; maintains circulating platelets in a resting state by phosphorylating proteins in numerous platelet inhibitory pathways when in complex with NF-kappa-B (NFKB1 and NFKB2) and I-kappa-B-alpha (NFKBIA), but thrombin and collagen disrupt these complexes and free active PRKACA stimulates platelets and leads to platelet aggregation by phosphorylating VASP. Prevents the antiproliferative and anti-invasive effects of alpha-difluoromethylornithine in breast cancer cells when activated. RYR2 channel activity is potentiated by phosphorylation in presence of luminal Ca (2+), leading to reduced amplitude and increased frequency of store overload-induced Ca (2+) release (SOICR) characterized by an increased rate of Ca (2+) release and propagation velocity of spontaneous Ca (2+) waves, despite reduced wave amplitude and resting cytosolic Ca (2+) . PSMC5/RPT6 activation by phosphorylation stimulates proteasome. Negatively regulates tight junctions (TJs) in ovarian cancer cells via CLDN3 phosphorylation. NFKB1 phosphorylation promotes NF-kappa-B p50-p50 DNA binding. Involved in embryonic development by down-regulating the Hedgehog (Hh) signaling pathway that determines embryo pattern formation and morphogenesis. Prevents meiosis resumption in prophase-arrested oocytes via CDC25B inactivation by phosphorylation. May also regulate rapid eye movement (REM) sleep in the pedunculopontine tegmental (PPT) . Phosphorylates APOBEC3G and AICDA. Isoform 2 phosphorylates and activates ABL1 in sperm flagellum to promote spermatozoa capacitation. Phosphorylates HSF1; this phosphorylation promotes HSF1 nuclear localization and transcriptional activity upon heat shock (PubMed:21085490) .
Type
ELISA Kit
Applications
Enzyme-linked Immunosorbent assay-Sandwich
Detection Method
Colorimetric
Assay Principle
The test principle applied in this kit is Sandwich enzyme immunoassay. The microtiter plate provided in this kit has been pre-coated with an antibody specific to Protein Kinase, cAMP Dependent Catalytic Alpha (PRKACa) . Standards or samples are then added to the appropriate microtiter plate wells with a biotin-conjugated antibody specific to Protein Kinase, cAMP Dependent Catalytic Alpha (PRKACa) . Next, Avidin conjugated to Horseradish Peroxidase (HRP) is added to each microplate well and incubated. After TMB substrate solution is added, only those wells that contain Protein Kinase, cAMP Dependent Catalytic Alpha (PRKACa), biotin-conjugated antibody and enzyme-conjugated Avidin will exhibit a change in color. The enzyme-substrate reaction is terminated by the addition of sulphuric acid solution and the color change is measured spectrophotometrically at a wavelength of 450nm +/- 10nm. The concentration of Protein Kinase, cAMP Dependent Catalytic Alpha (PRKACa) in the samples is then determined by comparing the O.D. of the samples to the standard curve.
Assay Protocol
Reconstitution & Storage Instructions Western Blotting/Immunoblotting (WB/IB) Protocol Immunohistochemistry (IHC) Protocol Immunocytochemistry (ICC) Protocol Enzyme-Linked ImmunoSorbent Assay (ELISA) Protocol Blocking Peptide Competition Protocol (BPCP) Immunoprecipitation (IP) Protocol Antibody Array (AA) Protocol Reconstitution & Storage Instructions Reconstitution & Storage Instructions Western Blotting/Immunoblotting (WB/IB) Protocol Western Blotting/Immunoblotting (WB/IB) Protocol Immunohistochemistry (IHC) Protocol Immunohistochemistry (IHC) Protocol Immunocytochemistry (ICC) Protocol Immunocytochemistry (ICC) Protocol Enzyme-Linked ImmunoSorbent Assay (ELISA) Protocol Enzyme-Linked ImmunoSorbent Assay (ELISA) Protocol Blocking Peptide Competition Protocol (BPCP) Blocking Peptide Competition Protocol (BPCP) Immunoprecipitation (IP) Protocol Immunoprecipitation (IP) Protocol Antibody Array (AA) Protocol Antibody Array (AA) Protocol
Sample Type
Tissue homogenates, cell lysates and other biological fluids
Detection Range
0.156-10ng/mL
Sensitivity
< 0.058ng/mL
Reconstitution
2°C to 8°C|-20°C
Components
Component Amount PRKACA Microplate 96 Wells (12 x 8 Well strips) PRKACA Lyophilized Standard 2 100X PRKACA HRP-Detector Anitbody 1 x 120 uL 100X Avidin-HRP Conjugate 1 x 120 uL Standard Diluent 1 x 20 mL Detector Antibody Diluent 1 x 12 mL Conjugate Diluent 1 x 12 mL 30X Wash Buffer 1 x 20 mL TMB Substrate 1 x 2 mL Stop Solution 1 x 20 mL Component Amount Component Amount PRKACA Microplate 96 Wells (12 x 8 Well strips) PRKACA Microplate 96 Wells (12 x 8 Well strips) PRKACA Lyophilized Standard 2 PRKACA Lyophilized Standard 2 100X PRKACA HRP-Detector Anitbody 1 x 120 uL 100X PRKACA HRP-Detector Anitbody 1 x 120 uL 100X Avidin-HRP Conjugate 1 x 120 uL 100X Avidin-HRP Conjugate 1 x 120 uL Standard Diluent 1 x 20 mL Standard Diluent 1 x 20 mL Detector Antibody Diluent 1 x 12 mL Detector Antibody Diluent 1 x 12 mL Conjugate Diluent 1 x 12 mL Conjugate Diluent 1 x 12 mL 30X Wash Buffer 1 x 20 mL 30X Wash Buffer 1 x 20 mL TMB Substrate 1 x 2 mL TMB Substrate 1 x 2 mL Stop Solution 1 x 20 mL Stop Solution 1 x 20 mL
NCBI Gene Symbol
PRKACA
Protein Name
CAMP-dependent protein kinase catalytic subunit alpha
Gene Name URL
PRKACA
Nucleotide Accession Number
NM_001304349.1