Example Protocol

Example Protocol Overview

The following steps provide an overview of an example In‑Cell Western Assay.

More detailed protocols and preparation instructions for solutions are provided in the pack inserts for CellTag™ 700 Stain Kits I and II and CellTag 520 Stain Kits III and IV.

  1. Seed cells - See Supplemental Material and Frequently Asked Questions and Supplemental Material and Frequently Asked Questions for more information.

  2. Cell treatment (optional)

  3. Fix cells - See Fixation and Permeabilization for more information.

  4. Permeabilize cells - See Fixation and Permeabilization for more information.

  5. Block cells - See Blocking Buffer Evaluation for more information.

  6. Prepare primary antibody solution

  7. Incubate primary antibody solution

  8. Wash plate

  9. Prepare secondary antibody solution and cell stain

  10. Incubate secondary antibody solution and cell stain

  11. Wash plate

  12. Acquire image - See licor.com/AcquisitionQuickStart for more information about using LI‑COR® Acquisition Software and an Odyssey Imager to acquire multiwell plate images.

    For more information about using Image Studio™ Software, see licor.com/bio/image-studio/resources.

After completing the protocol, analyze your plate image in Empiria Studio® Software using an appropriate Plate Template. In a Plate Template, you designate which wells will contain which samples and controls. During analysis, choose the multiwell plate image to analyze and the Plate Template to use for analysis. Wells on the image that correspond to wells designated on the Plate Template will be quantified. Statistical values will be calculated from the quantification values. See licor.com/AnalysisQuickStart for more information.