C3 ELISA Kit

Catalog No. ABIN955825

Pig C3 ELISA Kit, Colorimetric assay for quantification of Pig C3.

Quick Overview for C3 ELISA Kit (ABIN955825)

Target: C3 (Complement Component 3 (C3))

Reactivity: Pig

Detection Method: Colorimetric

Method Type: Sandwich ELISA

Detection Range: 62.5-4000 ng/mL

Application: ELISA

Product Details C3 ELISA Kit

Minimum Detection Limit: 62.5 ng/mL

Purpose: The kit is a sandwich enzyme immunoassay for the in vitro quantitative measurement of pig C3 in serum, plasma and other biological fluids.

Analytical Method: Quantitative

Specificity: This assay has high sensitivity and excellent specificity for detection of pig C3. No significant cross- reactivity or interference between pig C3 and analogues was observed. Note: Limited by current skills and knowledge, it is impossible for us - complete the cross-reactivity detection between pig C3 and all the other analogues, therefore, cross reaction may still exist.

Sensitivity: The minimum detectable dose of pig C3 is typically less than 15.8 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 C3. Calibrators or samples are then added to the appropriate microtiter plate wells with a biotin-conjugated polyclonal antibody preparation specific for C3. 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 C3, 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 +/- 10 nm. The concentration of C3 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 4, 000 ng/mL, 2, 1000 ng/mL, 500 ng/mL, 250 ng/mL, 125 ng/mL, 62.5 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 on log-log graph paper, with C3 concentration on the y-axis and absorbance on the x-axis. Draw the best fit straight line through the calibrator points and it can be determined by regression analysis. Using some plot software is also recommended. 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 for C3

Target: C3 (Complement Component 3 (C3))

Alternative Name: Complement C3

Pathways: Complement System, Regulation of Leukocyte Mediated Immunity, Positive Regulation of Immune Effector Process, Regulation of G-Protein Coupled Receptor Protein Signaling