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
Protocol Outline
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.
Storage/Stability
The entire kit may be stored at -20°C for up to 1 year from the date of shipment. Avoid repeated freeze-thaw cycles. The kit may be stored at 4°C for up to 6 months. For extended storage, it is recommended to store at -80°C.
