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Human MAP4K4(Mitogen-activated Protein Kinase 4) ELISA Kit
ELK0209
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48 T $412.00
96 T $588.00
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Overview

Product name: Human MAP4K4(Mitogen-activated Protein Kinase 4) ELISA Kit
Reactivity: Human
Alternative Names: HPK/GCK-like kinase HGK; MAPK/ERK kinase kinase kinase 4; MEK kinase kinase 4; MEKKK 4; Nck-interacting kinase
Assay Type: Sandwich
Sensitivity: 12.4 pg/mL
Standard: 3000 pg/mL
Detection Range: 46.88-3000 pg/mL
Sample Type: serum, plasma, tissue homogenates, cell lysates, cell culture supernates and other biological fluids
Assay Length: 3.5h
Research Area: Enzyme & Kinase;
Uniprot ID: O95819
Test 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 Human MAP4K4. Standards or samples are added to the appropriate microtiter plate wells then with a biotin-conjugated antibody specific to Human MAP4K4. 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 Human MAP4K4, 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 Human MAP4K4 in the samples is then determined by comparing the OD of the samples to the standard curve.

Standard curve

Concentration (pg/mL) OD Corrected OD
3000.00 2.153 2.072
1500.00 1.593 1.512
750.00 1.186 1.105
375.00 0.871 0.790
187.50 0.468 0.387
93.75 0.325 0.244
46.88 0.217 0.136
0.00 0.081 0.000

Precision

Intra-assay Precision (Precision within an assay)：CV%<8%

Three samples of known concentration were tested twenty times on one plate to assess intra-assay precision.

Inter-assay Precision (Precision between assays)：CV%<10%

Three samples of known concentration were tested in forty separate assays to assess inter-assay precision.

Recovery

Matrices listed below were spiked with certain level of recombinant MAP4K4 and the recovery rates were calculated by comparing the measured value to the expected amount of MAP4K4 in samples.
Matrix Recovery range Average
serum(n=5) 80-97% 88%
EDTA plasma(n=5) 78-92% 85%
Heparin plasma(n=5) 90-105% 97%

Linearity

The linearity of the kit was assayed by testing samples spiked with appropriate concentration of MAP4K4 and their serial dilutions. The results were demonstrated by the percentage of calculated concentration to the expected.
Matrix 1:2 1:4 1:8 1:16
serum(n=5) 98-103% 86-92% 87-93% 93-102%
EDTA plasma(n=5) 85-92% 79-96% 89-101% 85-97%
Heparin plasma(n=5) 86-94% 93-102% 89-97% 85-94%