Immunocytochemistry (ICC) is a technique for the visualization of proteins and peptides in cells using biomolecules capable of binding the protein of interest. Usually the biomolecule is an antibody that is linked to a reporter, e.g. a fluorophore, fluorescent dye, or enzyme. The reporter will give rise to a signal, e.g. fluorescence or color from an enzyme reaction, which is then detectable by a microscope.
Immunocytochemistry is often performed in four sequential steps:
- Cell seeding on a solid support
- Immunostaining of cells: cells are fixed, permeabilized, and stained with antibodies.
- Imaging: cells are visualized using a microscope and images acquired
- Image analysis
There are two different immunocytochemistry assay which include indirect and direct immunocytochemistry technique.
Direct immunocytochemistry assay involves the use of a detectable tag (e.g., fluorescent molecule, gold particles, etc.,) directly to the antibody that is then allowed to bind to the antigen (e.g., protein) in a cell. This method is rapid and specific.
Indirect immunocytochemistry assay involves antigen binding by a primary antibody which is then amplified by use of a secondary antibody which binds to the primary antibody. This method is more laborious and time-consuming, and a risk of non-specific binding of the secondary antibody.
Lightning-Link® for direct antibody labeling
Expedeon provides Lightning-Link® innovative technology that enables direct labelling of antibodies or proteins for use in immunoassay applications like Immunocytochemistry. The simplification of the conjugation process effectively circumvents issues like low sensitivity.
Expedeon Lightning-Link® product range includes kits for labelling antibodies with fluorescent dyes, fluorescent proteins, allowing quick and easy production of antibody conjugates. To produce the conjugate, the antibody is simply pipetted into a vial of lyophilized Lightning-Link® mixture containing the label of interest, incubated, and is then ready for use.
The Benefits of Lightning-Link® include:
- Require only 30 seconds hands-on time
- 100% recovery of materials
- The technology is fully scalable from 10ug to 1 g or more
- Stringently QC tested for consistent high quality
- Excellent batch-to-batch reproducibility.
The use of directly labelled primary antibodies demonstrates a similar level of staining to that which is produced when labeled secondaries are used.
To find out more about Lightning-Link®, why not watch our video?
Using Lightning-Link® conjugates for Immunocytochemistry – references Directly labeling primary antibodies using Lightning-Link® kits produces conjugates that can be used in a variety of scientific applications. Take a look at some of the published research in the table below that has used Lightning-Link® technology to produce conjugates for use within Immunocytochemistry… Title Pubmed ID Label…View
Download a Free Copy Lightning-Link® fluorescent antibody and protein labeling kits allow you to covalently label any antibody in under 20 minutes with only 30 seconds hands-on time. We currently have over 35 labels in our range of fluorescent dyes. This guide provides a general overview of each fluorochrome in our range including their properties,…View
Antibodies are widely employed in the quantification of antigens in complex biological samples. Using techniques such as Western blotting, ELISA, and immunohistochemistry researchers are able to measure a single antigen, or perhaps a limited number of antigens, in each sample. In the post-genomics era, advances in multiplex immunoassay technologies now allow scores or even hundreds…View
Expedeon specializes in easy to use bioconjugation kits which enable the direct labeling of antibodies or proteins with enzymes, fluorescent labels, biotin, streptavidin, gold nanoparticles, latex beads or oligonucleotides. Unfortunately, many antibodies are provided in buffers which contain additives that are incompatible with labeling technologies, making purification a key consideration prior to carrying out any…View
Download a free copy! Antibodies are used to detect and quantify antigens in techniques such as flow cytometry, ELISA, western blotting, immunohistochemistry and lateral flow. The antibody that binds to the antigen is called a ‘primary antibody’ and it confers specificity to the assay. Most types of immunoassays also incorporate a ‘label’ whose purpose is…View
Stop using secondary antibodies in immunoassays! Save time and materials by labeling your primary antibodies in under 20 min. For use in ELISA, western blotting, IHC, flow cytometry and many other applications. Scientists love our Lightning-Link® kits because of the remarkable simplicity of the technology. Now they can open a…View
**This webinar was produced by Innova Biosciences which is now fully integrated with their company, Expedeon Ltd., and have taken the Expedeon Ltd name and logo. Dr Nick Gee, the CEO and CSO of Expedeon, explains the benefits of using directly conjugated antibodies and the various approaches and techniques…View