This ELISA uses an antibody pair and standard that have been validated for multiplex array products, but not for the 96-well plate ELISA format. Thus, this ELISA requires a 4 - 7 week development phase, which RayBiotech begins after receiving an order for the kit. After development, the kit will have passed RayBiotech's ELISA quality control tests, and will thereafter have a typical 1 - 2 day lead time for orders placed.
The antibody pair provided in this kit recognizes Canine Neighbor of Punc E11 (IGDCC4)
Number of Targets Detected:
Method of Detection:
1, 2, or 5 x 96-Well Strip Microplate Kit
- Strip plates and additional reagents allow for use in multiple experiments
- Quantitative protein detection
- Establishes normal range
- The best products for confirmation of antibody array data
- Pre-Coated 96-well Strip Microplate
- Wash Buffer
- Stop Solution
- Assay Diluent(s)
- Lyophilized Standard
- Biotinylated Detection Antibody
- Streptavidin-Conjugated HRP
- TMB One-Step Substrate
Other Materials Required
- Distilled or deionized water
- Precision pipettes to deliver 2 µl to 1 µl volumes
- Adjustable 1-25 µl pipettes for reagent preparation
- 100 µl and 1 liter graduated cylinders
- Tubes to prepare standard and sample dilutions
- Absorbent paper
- Microplate reader capable of measuring absorbance at 450nm
- Log-log graph paper or computer and software for ELISA data analysis
- Prepare all reagents, samples and standards as instructed in the manual.
- Add 100 µl of standard or sample to each well.
- Incubate 2.5 h at RT or O/N at 4°C.
- Add 100 µl of prepared biotin antibody to each well.
- Incubate 1 h at RT.
- Add 100 µl of prepared Streptavidin solution to each well.
- Incubate 45 min at RT.
- Add 100 µl of TMB One-Step Substrate Reagent to each well.
- Incubate 30 min at RT.
- Add 50 µl of Stop Solution to each well.
- Read at 450 nm immediately.
May be stored for up to 6 months at 2° to 8°C from the date of shipment. Standard (recombinant protein) should be stored at -20°C or -80°C (recommended at -80°C) after reconstitution. Opened Microplate Wells or reagents may be store for up to 1 month at 2° to 8°C. Return unused wells to the pouch containing desiccant pack, reseal along entire edge. Note: the kit can be used within one year if the whole kit is stored at -20°C.