IHC and imaging antibodies

IHC antibodies are used to stain proteins and other analytes in paraffin-embedded and frozen tissue sections, as well as in cultured cells. We have spent over 25 years developing and validating antibodies in IHC and cell imaging. Our antibodies have been cited in over 75,000 scientific publications, including numerous imaging studies.

IHC and imaging antibodies, sample preparation, fixation, antigen retrieval, and staining

When using IHC antibodies, a complexity arises from the fact that not all antibodies can be used on samples preserved with the same fixation methods, and some antibodies require antigen retrieval treatments before the primary antibody can be applied. Some samples also require different degrees of permeabilization. These treatments are all needed to enable the antigen to expose or unmask the target epitopes that may have been obscured or altered during fixation. These steps must be carefully optimized to ensure the primary antibody can effectively bind to its target.

Developing and validating antibodies for imaging

Creating antibodies for IHC and other imaging methods like fluorescence staining isn’t just about binding. It’s about making sure the target antigen is visible and accessible in the sample.

We design immunogens with care, considering their position in the folded protein and using recombinant proteins where possible. This helps us create antibodies that are more likely to bind the right target.

Once developed, we screen candidate antibodies using control cell lines and healthy or diseased tissues. We look closely at how they localize in cells and tissues to confirm specificity and refine sample prep methods that best reveal the antigen.

To strengthen validation, we test IHC antibodies across techniques like flow cytometry, ELISA, and western blotting. We also use knockout cell lines to provide clear evidence of antibody specificity.

Conjugated antibodies for IHC and imaging

There are two methods for detecting antibody binding in IHC and cell staining. You can use a secondary antibody that’s conjugated to a label like an enzyme, fluorescent dye, or biotin. Or you can use a primary antibody that’s directly conjugated to the label.

Secondary antibodies offer signal amplification and flexibility. You can use the same unconjugated primary antibody across different methods. Directly conjugated antibodies simplify the protocol, reduce variability, and save time.

We offer both carrier-free antibodies ready for conjugation and a wide selection of pre-conjugated antibodies with fluorescent dyes, proteins, enzymes, and other labels.

Antibodies for multiplex IHC and multicolor imaging

Looking at just one protein isn’t always enough to understand complex biology. Imaging multiple proteins helps, but running separate experiments uses up valuable samples.

Multiplex staining enables the analysis of multiple targets simultaneously, saving time and preserving samples. The key is using labels that can be clearly distinguished, and antibodies that are compatible with those labels.

There are a few ways to do this:

• Use primary antibodies from different species, paired with matching secondary antibodies, like mouse, rabbit, and goat.

• Use directly conjugated primary antibodies labeled with fluorescent dyes or enzymes.

We support both approaches, helping you get more insight from every sample.

There are also more advanced methods for multiplex IHC. These include sequential staining with dye bleaching, metal labeling combined with mass spectrometry, and oligo-based antibody labeling. Each approach offers new ways to expand the number of targets you can analyze in a single sample.

Featured products