Components
Pre-coated 96 well plate
Standard
Assay Diluent concentrate
Biotinylated Detection Antibody
SAV-HRP
Wash Buffer
Chromogen
Stop Solution
Adhesive Plate Covers
Material not included
Precision pipette, disposable 5-1000µL plastic tip, 5-15m plastic pipette
A two-liter glass or plastic container to prepare wash buffer
Automatic 96-well dish washer
1.5mL polypropylene or polyethylene tube prep standard
Disposable reagent reservoir
15mL plastic tube to prepare streptavidin-hrp solution
Standard ELISA reader for absorbance measurements at 450nm and 550nm.
Graphics paper or computerized curve fitting statistical software package
Sample Volume
1 µL to 50 µL
Assay Time
Less than 3 hours
Reagent Preparation
1. Prepare Wash Buffer
Allow wash buffer concentrate to reach room temperature and mix to redissolve any precipitated salts. Dilute Wash Buffer Concentrate into deionized or distilled water. Store the concentrate and 1X wash buffer in the refrigerator.
2. Prepare the thinner
The test diluent should be diluted with deionized or distilled water before use.
3. Preparation of Biotin Conjugation
Spin the biotin conjugate briefly before use. Gently stir up and down with a pipette. Collect samples in pyrogen/endotoxin-free tubes. If samples cannot be tested immediately after collection, freeze samples. Freeze samples to avoid multiple freeze-thaw cycles.
4. Pre-dilute samples
For different types of samples, different dilution solutions should be used for dilution.
5. Prepare 1X Streptavidin-HRP Solution
Quickly swirl Streptavidin-HRP before use and mix gently up and down with a pipette as a precipitate may form during storage. Do not store the diluted solution for future use.
6. Dilution Standards
Dilute standards using glass or plastic tubes. Simply spin off a vial of lyophilized standards.
Assay Procedure
1. Sample Incubation
2. Plate Washing
3. Biotinylated Antibody Reagent Incubation
4. Streptavidin-HRP Solution Preparation and Incubation
5. Substrate Incubation and Stop Step
6. Absorbance Measurement
Note: Evaluate the plate within 30-45 minutes of stopping the reaction.
Calculation of Results
Calculate results using graph paper or curve-fitting statistical software. The human CCL5 amount in each sample is determined by interpolating from the absorbance value (Y axis) to human CCL5 concentration (X axis) using the standard curve.
Assay Precision
51.2-2000 pg/mL
Precaution of Use
All products are supplied for research and laboratory use only.
Handling Advice
All blood components and biological materials should be treated with precautions as potentially hazardous. Strictly follow the management principles of the Centers for Disease Control and Prevention, Occupational Safety and Health Administration for handling and disposing of infectious agents.
Storage
The ELISA Kits are shipped at 2 to 8°C. Upon receipt, store the kits at 2 to 8°C in dark.
Expiry Date
Stability If properly stored, all components are stable for up to 12 months. For expiry dates for the entire kit, see the kit label. The expiration date of each ingredient is shown on the bottle label. The expiration date of a kit ingredient is guaranteed only if the ingredient is properly stored and, in the event of repeated use of an component, the reagent will not be contaminated by the first treatment.
Note
Each production lot of this ELISA kit is quality tested to meet criteria such as sensitivity, specificity, precision and lot-to-lot consistency. See the manual for more information on validation.
Datasheet