Secondary antibodies are a vital reagent for any fluorescent detection technique, including flow cytometry and imaging. That’s because while primary antibodies are important for detecting a protein of interest, it’s the secondary antibodies that provide the fluorescent signal that allows for the detection of binding of the primary antibody. It’s therefore imperative that scientists have access to the appropriate secondary reagents to bind the species of the primary antibodies used to target their proteins of interest. But can sometimes be challenging to find the required secondary antibody, especially when looking for one that targets a specific isotype or subclass of primary antibody. This becomes a particular challenge when combining very similar primary antibodies in a single study – for example, when using two rat-derived antibodies that only differ because one is an IgG and the other an IgM.
Luckily, Jackson ImmunoResearch offers a comprehensive profile of secondary antibodies that target a variety of specific primary antibodies and are conjugated to a diverse array of fluorophores for detection by microscopy or flow cytometry.
In my research, I order antibodies from “JIR” because the combination of targets that I am interested in investigating by microscopy often require using antibodies derived from the same species. For example, I frequently use a rat IgG and a rat IgM primary antibody to detect two vascular targets important to my research. In a more recent study, I’ve needed to co-stain with a mouse IgG1 and a mouse IgG2a antibody when investigating a specific subset of memory T cells. In both of these cases, I’m not able to use the species-specific secondary reagents that are commonly sold by several vendors; I need something that targets particular isotypes or even subclasses of antibodies. JIR sells high-quality, low-cost secondary reagents that allow me to use these primary antibody panels for immunofluorescence microscopy.
Because of the range of colors offered, similar reagents from JIR can be used in flow cytometric or multispectral imaging analyses; for example, horseradish peroxidase (HRP)-conjugated reagents are useful for many signal amplification techniques, and secondary reagents that emit at wavelengths of 405, 421, and 675 can be used in flow cytometry.
JIR allows you to browse secondary reagents by species, antibody isotype or subclass, color, or experimental application. And all of the reagents are very reasonably priced! Almost all of the secondary reagents cost less than $200 for 0.5 mg, with many in the $100 range depending on the antibody specificity or conjugate.
A great source for JIR products is Quartzy - you can browse and buy JIR products in the Quartzy Shop and since they have so many other suppliers you can often consolidate orders on top of getting their negotiated pricing. This means that combining similar primary antibodies is not only feasible scientifically, but also financially, thanks to the products offered by JIR.
Quartzy is the world’s No. 1 lab management platform. We help scientists easily organize orders, manage inventory, and save money. We’re free and always will be. Visit Quartzy.com or reach out at firstname.lastname@example.org.
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Aliyah is a PhD candidate at the University of Pittsburgh, where she studies cancer immunology. She is also an advocate for science communication. You can find her on Twitter @desabsurdites and on her blog at http://isntthatgrad.wordpress.com/.