RayBio® Rat Tumor Necrosis Factor alpha (TNF-alpha) ELISA Kit For Cell & Tissue 1 x 96 well

• P16599

• 103694380
• 24835

• Rat

• 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

Kit Components

• 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
• Cell Lysis Buffer

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

Protocol Outline

1. Prepare all reagents, samples and standards as instructed in the manual.
2. Add 100 µl of standard or sample to each well.
3. Incubate 2.5 h at RT or O/N at 4°C.
4. Add 100 µl of prepared biotin antibody to each well.
5. Incubate 1 h at RT.
6. Add 100 µl of prepared Streptavidin solution to each well.
7. Incubate 45 min at RT.
8. Add 100 µl of TMB One-Step Substrate Reagent to each well.
9. Incubate 30 min at RT.
10. Add 50 µl of Stop Solution to each well.
11. Read at 450 nm immediately.