CF-B ELISA Kit

Catalog No. ABIN955831

Product Details

Target: CF-B

Reactivity: Mouse

Detection Method: Colorimetric

Method Type: Sandwich ELISA

Detection Range: 6.25-400 ng/mL

Minimum Detection Limit: 6.25 ng/mL

Application: ELISA

Purpose: The kit is a sandwich enzyme immunoassay for the in vitro quantitative measurement of mouse CF-B in plasma.

Analytical Method: Quantitative

Specificity: This assay has high sensitivity and excellent specificity for detection of mouse CF-B. No significant cross- reactivity or interference was observed.

Sensitivity: The minimum detectable dose of mouse CF-B is typically less than 1.94 ng/mL.
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 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 CF-B. Calibrators or samples are then added to the appropriate microtiter plate wells with a biotin-conjugated polyclonal antibody preparation specific for CF-B. Next, Avidin conjugated to Horseradish Peroxidase (HRP) is added to each microplate well and incubated. Then a TMB substrate solution is added to each well. Only those wells that contain CF-B, 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 CF-B 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 strip 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. De-ionized 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 400 ng/mL, 200 ng/mL, 100 ng/mL, 50 ng/mL, 25 ng/mL, 12.5 ng/mL, 6.25 ng/mL.

Plate: Pre-coated

Calculation of Results:

Average the duplicate readings for each calibrator, control, and samples 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 CF-B concentrations versus the log of the O.D. and the best fit line can be determined by regression analysis. It is recommended to use some related software to do this calculation. This procedure will produce an adequate but less precise fit of the data. 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 the reagents should be kept according to the labels on vials. The Calibrator, Detection Reagent A, Detection Reagent B and the 96-well strip plate should be stored at -20° C upon being received. 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 shown, provided it is stored as prescribed above.

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

Target Details

Target: CF-B