Rabbit GRO alpha/CXCL1 ELISA Kit

1. Target Protein Microplate – 96 wells (12 strips x 8 wells) coated with anti-target protein
2. Target Protein Standard – 2 vials
3. Target Protein Detection Antibody – 2 vials of biotinylated anti-target protein
4. Wash Buffer – 25 ml of 20X concentrated solution
5. HRP-Streptavidin – 200 µl of concentrated HRP-conjugated streptavidin
6. TMB One-Step Substrate Reagent – 12 ml
7. Stop Solution – 8 ml of 0.2 M sulfuric acid
8. Assay Diluent(s) – diluents for samples, standard and HRP-Streptavidin

Other materials and equipment required:

The Assay Genie Rabbit GRO alpha/CXCL1 ELISA Kit (SBRS7295) will require other equipment and materials to carry out the assay. Please see list below for further details.

• Microplate reader capable of measuring absorbance at 450nm
• Precision pipettes to deliver 2 ul to 1 ml volumes
• Adjustable 1-25 ml pipettes for reagent preparation
• 100 ml and 1 liter graduated cylinders
• Tubes to prepare standard or sample dilutions
• Absorbent paper
• Distilled or deionized water
• Log-log graph paper or computer and software for ELISA data analysis

1. Bring all reagents and samples to room temperature (18 - 25°C) and prepare all reagents, samples and standards as instructed.
2. Add 100 ul standard or sample to each well.
3. Incubate 2.5 h at RT with gentle shaking.
4. Wash 4 times with 1X Wash Buffer (300 ul per well).
5. Add 100 ul prepared biotinylated detection antibody to each well.
6. Incubate 1 h at RT with gentle shaking.
7. Wash 4 times with 1X Wash Buffer.
8. Add 100 ul prepared streptavidin solution to each well.
9. Incubate 45 min at RT with gentle shaking.
10. Wash 4 times with 1X Wash Buffer.
11. Add 100 ul TMB One-Step Substrate Reagent to each well.
12. Incubate 30 min at RT in the dark with gentle shaking.
13. Add 50 ul Stop Solution to each well.
14. Read plate at 450 nm immediately.