C5A ELISA Kit

Catalog No. ABIN955827

Product Details C5A ELISA Kit

Target: C5A (C5a) (Complement Component 5a (C5a))

Reactivity: Mouse

Detection Method: Colorimetric

Method Type: Sandwich ELISA

Detection Range: 78-5000 pg/mL

Minimum Detection Limit: 78 pg/mL

Application: ELISA

Purpose: The kit is a sandwich enzyme immunoassay for the in vitro quantitative measurement of mouse C5a in serum, plasma, tissue homogenates, cell culture supernates and other biological fluids.

Analytical Method: Quantitative

Specificity: This assay has high sensitivity and excellent specificity for detection of mouse C5a. No significant cross- reactivity or interference between mouse C5a and analogues was observed. Note: Limited by current skills and knowledge, it is impossible for us - complete the cross-reactivity detection between mouse C5a and all the other analogues, therefore, cross reaction may still exist.

Sensitivity: The minimum detectable dose of mouse C5 a is typically less than 23 pg/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 C5a. Calibrators or samples are then added to the appropriate microtiter plate wells with a biotin-conjugated polyclonal antibody preparation specific for C5a. 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 C5a, 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 C5a 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 5, 000 pg/mL, 2, 500 pg/mL, 1250 pg/mL, 625 pg/mL, 312 pg/mL, 156 pg/mL, 78 pg/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 C5a 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 C5A

Target: C5A (C5a) (Complement Component 5a (C5a))

Abstract: C5a Products

Synonyms: C5a ELISA Kit, C5b ELISA Kit, CPAMD4 ELISA Kit, Hc ELISA Kit, RGD1561905 ELISA Kit, complement C5 ELISA Kit, C5 ELISA Kit

Pathways: Complement System, Carbohydrate Homeostasis