ELISA Kit for Creatine Kinase, Muscle (CKM)Homo sapiens (Human)Sandwich ELISA

CK-MM; CKMM; CK3; CK-M; MM-CK; M-CK; Creatine kinase M-type, N-terminally processed

  • Product No.SEA109Hu
  • Organism SpeciesHomo sapiens (Human)
  • Sample Typeserum, plasma, tissue homogenates, cell lysates, cell culture supernates and other biological fluids
  • Test MethodDouble-antibody Sandwich
  • Assay Length3h
  • Detection Range1.56-100ng/mL
  • SensitivityThe minimum detectable dose of this kit is typically less than 0.59ng/mL.
  • Specificity This assay has high sensitivity and excellent specificity for detection of Creatine Kinase, Muscle (CKM).
    No significant cross-reactivity or interference between Creatine Kinase, Muscle (CKM) and analogues was observed.
  • ApplicationsEnzyme-linked immunosorbent assay for Antigen Detection
  • DownloadInstruction Manual
  • Format48T96T96T*596T*1096T*100
  • FOBUS$ 441US$ 630US$ 2835US$ 5355US$ 44100
  • ELISA Kit for Creatine Kinase, Muscle (CKM)Package and Components
  • ELISA Kit for Creatine Kinase, Muscle (CKM)Package and Components
  • ELISA Kit for Creatine Kinase, Muscle (CKM)Results demonstration
  • SEA109Hu.jpgTypical Standard Curve
  • CertificateISO9001: 2008, ISO13485: 2003 Registered

Recovery

Matrices listed below were spiked with certain level of recombinant Creatine Kinase, Muscle (CKM) and the recovery rates were calculated by comparing the measured value to the expected amount of Creatine Kinase, Muscle (CKM) in samples.

MatrixRecovery range (%)Average(%)
serum(n=5)80-9181
EDTA plasma(n=5)80-9582
heparin plasma(n=5)85-10391

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Creatine Kinase, Muscle (CKM) were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level Creatine Kinase, Muscle (CKM) were tested on 3 different plates, 8 replicates in each plate.
CV(%) = SD/meanX100
Intra-Assay: CV<10%
Inter-Assay: CV<12%

Linearity

The linearity of the kit was assayed by testing samples spiked with appropriate concentration of Creatine Kinase, Muscle (CKM) and their serial dilutions. The results were demonstrated by the percentage of calculated concentration to the expected.

Sample1:21:41:81:16
serum(n=5)94-104%85-94%78-89%86-95%
EDTA plasma(n=5)86-95%98-105%88-95%81-104%
heparin plasma(n=5)94-101%86-101%82-102%84-93%

Stability

The stability of kit is determined by the loss rate of activity. The loss rate of this kit is less than 5% within the expiration date under appropriate storage condition.
To minimize extra influence on the performance, operation procedures and lab conditions, especially room temperature, air humidity, incubator temperature should be strictly controlled. It is also strongly suggested that the whole assay is performed by the same operator from the beginning to the end.

Reagents and materials provided

ReagentsQuantityReagentsQuantity
Pre-coated, ready to use 96-well strip plate1Plate sealer for 96 wells4
Standard2Standard Diluent1×20mL
Detection Reagent A1×120µLAssay Diluent A1×12mL
Detection Reagent B1×120µLAssay Diluent B1×12mL
TMB Substrate1×9mLStop Solution1×6mL
Wash Buffer (30 × concentrate)1×20mLInstruction manual1

Assay procedure summary

1. Prepare all reagents, samples and standards;
2. Add 100µL standard or sample to each well. Incubate 1 hours at 37°C;
3. Aspirate and add 100µL prepared Detection Reagent A. Incubate 1 hour at 37°C;
4. Aspirate and wash 3 times;
5. Add 100µL prepared Detection Reagent B. Incubate 30 minutes at 37°C;
6. Aspirate and wash 5 times;
7. Add 90µL Substrate Solution. Incubate 15-25 minutes at 37°C;
8. Add 50µL Stop Solution. Read at 450nm immediately.

ELISA Kit for Creatine Kinase, Muscle (CKM)

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 Creatine Kinase, Muscle (CKM). Standards or samples are then added to the appropriate microtiter plate wells with a biotin-conjugated antibody specific to Creatine Kinase, Muscle (CKM). 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 Creatine Kinase, Muscle (CKM), 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 Creatine Kinase, Muscle (CKM) in the samples is then determined by comparing the O.D. of the samples to the standard curve.

Reference

  • Journal of InflammationAssessment of endothelium and inflammatory response at the onset of reperfusion injury in hand surgeryPubMed: 22583529
  • PLoS OneExtraocular Muscle Characteristics Related to Myasthenia Gravis SusceptibilityPubMed: PMC3568149
  • Kidney Int. Fructokinase activity mediates dehydration-induced renal injuryPubmed: 24336030