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Catalog No: OKCD00537 (Formerly GWB-KBBWW7)
Size:96WELLS
Price: $675.00
SKU
OKCD00537
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Datasheets/ManualsPrintable datasheet for PAK4 ELISA Kit (Human) (OKCD00537)
Product Info
Predicted Species ReactivityHomo sapiens|Human
ApplicationEnzyme-linked Immunosorbent assay-Sandwich
ELISA Kit Detection MethodColorimetric
ELISA Kit Duration3h
ELISA Kit Linearity
Sample1:2 Dilution1:4 Dilution1:8 Dilution1:16 Dilution
Serum (n=5)88-97%93-101%81-88%86-93%
EDTA Plasma (n=5)79-101%96-103%88-95%99-105%
Heparin Plasma (n=5)82-89%80-92%95-105%80-101%
ELISA Kit PrincipleThe 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 p21 Protein Activated Kinase 4 (PAK4). Standards or samples are then added to the appropriate microtiter plate wells with a biotin-conjugated antibody specific to p21 Protein Activated Kinase 4 (PAK4). 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 p21 Protein Activated Kinase 4 (PAK4), 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 p21 Protein Activated Kinase 4 (PAK4) in the samples is then determined by comparing the O.D. of the samples to the standard curve.
ELISA Kit Range0.156-10ng/mL
ELISA Kit Recovery
MatrixRecovery Range (%)Average (%)
serum(n=5)94-10397
EDTA plasma(n=5)87-10399
heparin plasma(n=5)84-10193
ELISA Kit ReproducibilityIntra-assay Precision (Precision within an assay): 3 samples with low, middle and high level p21 Protein Activated Kinase 4 (PAK4) were tested 20 times on one plate, respectively. Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level p21 Protein Activated Kinase 4 (PAK4) were tested on 3 different plates, 8 replicates in each plate. CV(%) = SD/meanX100 Intra-Assay: CV<10% Inter-Assay: CV<12%
ELISA Kit Component
ComponentAmount
Anti-PAK4 Microplate96 Wells (12 x 8 Well Strips)
PAK4 Lyophilized Standard2 x 10 ng
100X Biotinylated PAK4 Detector Antibody120 uL
Avidin/HRP Conjugate120 uL
Standard Diluent1 x 20 mL
Detector Antibody Diluent1 x 12 mL
Conjugate Diluent1 x 12 mL
30X Wash Buffer1 x 20 mL
TMB Substrate1 x 9 mL
Stop Solution1 x 6 mL
Reconstitution and Storage2°C to 8°C|-20°C
Sample TypeSerum, plasma, tissue homogenates and other biological fluids.
Sensitivity< 0.065 ng/mL
SpecificityThis assay has high sensitivity and excellent specificity for detection of p21 Protein Activated Kinase 4 (PAK4). No significant cross-reactivity or interference between p21 Protein Activated Kinase 4 (PAK4) and analogues was observed.
Assay InfoAssay Methodology: Quantitative Sandwich Immunoassay
Gene SymbolPAK4
Gene Full Namep21 (RAC1) activated kinase 4
Alias Symbolsp21 protein (Cdc42/Rac)-activated kinase 4;p21(CDKN1A)-activated kinase 4;p21-activated kinase 4;protein kinase related to S. cerevisiae STE20, effector for Cdc42Hs;serine/threonine-protein kinase PAK 4.
NCBI Gene Id10298
Protein NameSerine/threonine-protein kinase PAK 4
Description of TargetSerine/threonine protein kinase that plays a role in a variety of different signaling pathways including cytoskeleton regulation, cell migration, growth, proliferation or cell survival. Activation by various effectors including growth factor receptors or active CDC42 and RAC1 results in a conformational change and a subsequent autophosphorylation on several serine and/or threonine residues. Phosphorylates and inactivates the protein phosphatase SSH1, leading to increased inhibitory phosphorylation of the actin binding/depolymerizing factor cofilin. Decreased cofilin activity may lead to stabilization of actin filaments. Phosphorylates LIMK1, a kinase that also inhibits the activity of cofilin. Phosphorylates integrin beta5/ITGB5 and thus regulates cell motility. Phosphorylates ARHGEF2 and activates the downstream target RHOA that plays a role in the regulation of assembly of focal adhesions and actin stress fibers. Stimulates cell survival by phosphorylating the BCL2 antagonist of cell death BAD. Alternatively, inhibits apoptosis by preventing caspase-8 binding to death domain receptors in a kinase independent manner. Plays a role in cell-cycle progression by controlling levels of the cell-cycle regulatory protein CDKN1A and by phosphorylating RAN.
Uniprot IDO96013
Protein Accession #NP_001014831
Nucleotide Accession #NM_001014831.2
Protein Size (# AA)591
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