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 27-Hydroxycholesterol.
Shipping Condition:
Ice packs
Assay Procedure Summary:
1. Prepare the reagents, standards, and samples needed for testing; 2. Aliquot 50 µl of diluted standard into the standard wells; 3. Aliquot 50 µl of Standard Diluent buffer into control (zero) well; 4. Add 50 µL/well sample to the microtiter plate; 5. Then add 50 µL of the prepared detection solution A, incubate 45 min at 37°C; 6. After washing the plate 3 times, add 100 µL of the prepared detection solution B and incubate 30 min at 37°C; 7. After washing the plate 5 times, add 50 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.
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.
ELISA Kit Components:
Pre-coated 96-Well Microplate
Standard
Diluent A
Diluent B
Wash Buffer
Detection Solution A
Detection Solution B
Standard Diluent Buffer
TMB Substrate
Stop Solution
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.
Standard Form:
Lyophilized
