Storage:
Short-term storage at 4℃, some reagents should be kept at -20℃ when not in use for a long time.
Detection Method:
Colorimetric
Specificity:
This ELISA kit has excellent specificity for the detection of Complement C4.
Shipping Condition:
Ice packs
Standard Form:
Lyophilized
Sample Preparation:
Serum: Collect the sample in a serum separation tube and place the tube in a vertical position overnight at 4°C. Then centrifuge at approximately 1000 × g for 20 minutes. Analyze the serum immediately or aliquot and store at -20°C or -80°C.
Plasma: Use anticoagulant to collect plasma. Centrifuge for 15 minutes at 1000 × g within 30 minutes of collection. Assay immediately or aliquot and store at -20°C or -80°C. Avoid hemolysis and high cholesterol samples.
Tissue homogenates: Rinse tissues with ice-cold PBS to remove the excess of blood. Weigh the tissue first, homogenize the tissue on ice with a homogenizer, and then ultrasonically process the cell suspension. Centrifuge the homogenates at 5000 × g for 5 min and collect the supernatant. Assay immediately or aliquot and store at -20°C.
Cross Activity:
It has been verified that this product has high specificity and sensitivity to Complement C4, and no significant cross-reactivity or interference with analogues was observed.
Assay Procedure Summary:
1. Prepare the reagents, standards, and samples needed for testing; 2. Aliquot 100 µl of diluted standard into the standard wells; 3. Aliquot 100 µl of standard diluent buffer into control (zero) well; 4. Add 100 µL/well sample to the microtiter plate, incubate 1 h at 37°C; 5. Then add 100 µL of the prepared detection solution A, incubate 1 h at 37°C; 6. After washing the plate 3 times, add 100 µL of the prepared detection solution B and incubate 90 min at 37°C; 7. After washing the plate 5 times, add 90 µL of substrate solution and incubate 10-20 min at 37°C; 8. Terminated by the addition of 50 µL stop solution, the absorbance was read at 450 nm by a microplate reader.
ELISA Kit Components:
Pre-coated 96-Well Microplate
Standard
Solution A
Solution B
Wash Buffer
Detection Solution A
Detection Solution B
Standard Diluent Buffer
TMB Substrate
Stop Solution
Precautions before Use:
The kits of this ELISA kit are not assembled, please assemble it before use.
Stop Solution:
The stop solution of this kit contains acid. Please pay attention to safe use and avoid direct contact.
Notice:
In order to reduce the influence of various variables on the experimental results during the experiment, you must strictly control the experimental conditions, and the entire experimental operation process should be completed by the same person.
