CKM ELISA Kit

Catalog No. ABIN955838

This Colorimetric ELISA kit is designed for the quantitative measurement of Rat CKM.

Quick Overview for CKM ELISA Kit (ABIN955838)

Target: CKM (Creatine Kinase, Muscle (CKM))

Reactivity: Rat

Detection Method: Colorimetric

Method Type: Sandwich ELISA

Detection Range: 7.8-500 U/L

Application: ELISA

Product Details CKM ELISA Kit

Minimum Detection Limit: 7.8 U/L

Purpose: This ELISA kit is a sandwich enzyme immunoassay for the in vitro quantitative measurement of Rat CK-M in serum, plasma and other biological fluids.

Analytical Method: Quantitative

Specificity: This assay has high sensitivity and excellent specificity for detection of Rat CK-M. No significant cross- reactivity or interference was observed.

Sensitivity: The minimum detectable dose of Rat CK-M is typically less than 2.1 U/L. The Sensitivity of this assay, or Lower Limit of Detection (LLD) was defined as the lowest protein concentration that could be differentiated from zero. It was determined by the mean O.D. value of 20 replicates of the zero calibrator plus three standard deviations.

Characteristics: The microtiter plate provided in this kit has been pre-coated with an antibody specific to CK-M. Calibrators and samples are then added to the appropriate microtiter plate wells with a biotin-conjugated polyclonal antibody preparation specific for CK-M. Next, Avidin conjugated to Horseradish Peroxidase (HRP) is added to each microplate well and incubated. Then the TMB substrate solution is added to each well. Only those wells that contain CK-M, biotin-conjugated antibody and enzyme-conjugated Avidin will exhibit a change in color. The enzyme-substrate reaction is terminated by the addition of a sulfuric acid solution and the color change is measured spectrophotometrically at a wavelength of 450 nm +/- 2 nm. The concentration of CK-M in the samples is then determined by comparing the O.D. of the samples to the calibration curve.

Components: Pre-coated, ready to use 96-well plate (1x)
Calibrator (lyophilized) (2x)
Calibrator Diluent (1 x 20 mL)
Detection Reagent A (1 x 120 µL)
Detection Reagent B (1 x 120 µL)
Assay Diluent A (2X concentrate) (1 x 6 mL)
Assay Diluent B (2X concentrate) (1 x 6 mL)
TMB Substrate (1 x 9 mL)
Stop Solution (1 x 6 mL)
Wash Buffer (30X concentrate) (1 x 20 mL)
Plate sealer for 96 wells (4x).

Material not included: 1. Microplate reader with 450 +/- 10 nm filter.
2. Precision single and multi-channel pipettes and disposable tips.
3. Eppendorf Tubes for diluting samples.
4. Deionized or distilled water.
5. Absorbent paper for blotting the microtiter plate.
6. Container for Wash Solution

Application Details

Comment:

The calibration curve concentrations used for the ELISA's were 500, 250, 125, 62.5, 31.2, 15.6, 7.8 U/L.

Plate: Pre-coated

Calculation of Results:

Average the duplicate readings for each calibrator, control, and sample and subtract the average zero calibrator optical density. Create a calibration curve by reducing the data using computer software capable of generating a four parameter logistic (4-PL) curve-fit. As an alternative, construct a calibration curve by plotting the mean absorbance for each calibrator on the x-axis against the concentration on the y-axis and draw a best fit curve through the points on the graph. The data may be linearized by plotting the log of the CK-M concentrations versus the log of the O.D. and the best fit line can be determined by regression analysis. This procedure will produce an adequate but less precise fit of the data. It is recommended to use some related software to do this calculation. If samples have been diluted, the concentration read from the calibration curve must be multiplied by the dilution factor.

Restrictions: For Research Use only

Handling

Storage: -20 °C

Storage Comment: All reagents should be stored according to their label. The Calibrators, Detection Reagent A, Detection Reagent B and the 96-well plate should be stored at -20° C upon receipt. The unused strips should be kept in a sealed bag with the desiccant provided to minimize exposure to damp air. Opened test kits will remain stable until the expiration date, provided they are stored as above.

Expiry Date: The expiry date is stated on the label.

Target Details for CKM

Target: CKM (Creatine Kinase, Muscle (CKM))

Alternative Name: CK-M