Preparing Cell or Tissue Lysates for ELISA & Multiplex Assay Kits
RayBiotech manufactures over 7,000 high fully validated, GMP-compliant ELISA kits. In this blog post we explain how to prepare cell or tissue lysates for ELISA Kits.
Preparation
Cell or tissue lysates for use with RayBio® ELISA kits can be prepared using most conventional methods, e.g. homogenization of cell or tissue in RayBio® Lysis Buffer. You may also use your own lysis buffer, such as RIPA or other formulations optimized for immunoprecipitation.
Please note the following guidelines on lysis buffer composition:
- Avoid using >0.1% SDS or other strongly denaturing detergents. In general, non-ionic detergents such as Triton X-100 or NP-40 are best, although zwitterionic detergents such as CHAPS, or mild ionic detergents such as sodium deoxycholate will work.
- Use no more than 2% v/v total detergent
- Avoid the use of sodium azide
- Avoid using >10 mM reducing agents, such as dithiothreitol or mercaptoethanols
We strongly recommend adding a protease inhibitor cocktail to the lysis buffer prior to homogenization. Most general biochemical supply companies including Roche, Sigma-Aldrich, Pierce, and Calbiochem stock a wide variety of these products. Since susceptibility to proteolytic cleavage and the type of proteases present in the lysate vary, we do not recommend a specific product. Instead, your choice of which combination of protease inhibitors to use should be based upon a literature search for your protein(s) of interest and/or tissue or cell type. Phosphatase inhibitors may be used but are not necessary unless the antibodies used in the kit specifically recognize phosphorylated forms of the protein.
Choices of the method for lysis and homogenization include glass-bead “smash,” douncing, freeze-thaw, sonication and crushing frozen tissue with a mortar and pestle, or even a combination of these. There is no best method for all sample types; your choice of method should be made following a brief search of the literature to see how samples similar to yours have been prepared in previous investigations.
7 Feedbacks on "Preparing Cell or Tissue Lysates for ELISA & Multiplex Assay Kits"
Supreme QA
July 30 2026 at 12:44 pmFor testing purpose- In the mouse cAMP ELISA kit, the result is represented in ng/mL. But in the published article the cAMP is measured in pmol or fmol. Since cAMP released in minor quantity it can be measured only in pmol or fmol. Could you please help me to understand this
- Hi I wanna to set an ELISA to evaluate binding ability of my scFv produced in E.coli to cell surface marker. As the target protein is a transmembrane, what should I do? I used to do in-cell ELISA, however even after fixation with 8% formaldehyde, many of the cells were detached from the wells during washing steps. My cells are not adherent. I prefer to work with cellular lysate, however, it has two problem: protease degradation and high background signal. can I use a denaturant to inactivate all peroxidases and protease? My target is a linear sequence of amino acids, so 3D or 2D structure of the target protein is not important. The important thing is that the transmembrane protein is compeletely released from membrane lipids and is able to be coated on the plate. So, can I use 8M urea in the lysis buffer? Can I boile the lysate?