Guinea pig antibodies offer unique benefits that make them valuable tools in research.
We want you to have as much freedom as possible when designing your experiments, which is why we’d like to introduce you to our range of primary antibodies raised in guinea pig. These antibodies, while not inherently more specific or sensitive than those from rabbits or mice, can offer you more options when designing multiplex staining and complex immunoprecipitation (IP) experiments.
Why Choose Guinea Pig Antibodies?
We’ve all used rabbit and mouse primary antibodies for so long that the guinea pig host has perhaps been overlooked by many. And that’s a shame because antibodies from guinea pigs are particularly useful in experimental setups that need multiple antibodies from different species to avoid cross reactivity in which your secondary antibody binds to the wrong primary antibody.
What’s also interesting at the antibody-generation level is that due to their unique phylogenetic bifurcation, guinea pigs have varying degree of homology with human proteins. If you think about immunogenicity, this is a great thing because with guinea pigs you can generate antibodies to some human proteins that you couldn’t do so in mice or rats. In addition, it is commonly accepted that guinea pig’s response to an antigen is much greater than that in mice or rats, so you need less antigen to get the process going.
But let’s look at the practical uses in the lab. At lot of the time, you need to move beyond the norm of one or two proteins of interest in your assay. There are occasions when you want to ramp it up to three or four or even more. At this point, you need to add more reliable antibodies from different hosts into the mix, such as guinea pig, rabbit, and mouse, so that when it comes to choosing a secondary antibody, can you be sure it only reacts with one of your primaries (assuming you want accurate and simultaneous visualization of multiple targets within a single sample, that is!). This is where having guinea pig antibodies available gives you more options to target more proteins without making a mess of things.
For IP and particularly co-IP experiments, one of the primary benefits of using antibodies from multiple species is to avoid cross-reactivity when you have multiple antibodies being used to pull down different proteins within a complex. By using antibodies from different hosts, you can easily pair secondary antibodies with each primary antibody without interference.
For example, you might use a guinea pig antibody to target a protein in conjunction with a rabbit antibody against another protein within the same complex. Using a dual-species approach can improve the precision of your pull-down since each antibody binds specifically to its target protein without any unwanted binding to the other components in the lysate.
Guinea Pig Antibodies in Action
In a study investigating the interaction of astrocytes and autoreactive immune cells in a multiple sclerosis (MS) model – experimental autoimmune encephalomyelitis (EAE), researchers used a suite of immunoassays to study the direct interaction of these cells when they are in close proximity to the inflamed CNS (1). The team used our guinea pig Anti-Connexin-43 Antibody (#ACC-201-GP) alongside our rabbit Anti-P2X7 Receptor Antibody (#APR-004) and mouse anti-CD4 to demonstrate the colocalization of astrocytic P2X7 receptor and connexin-43 with the CD4+ T cells in the spinal cord.
The team were able to show a dense colocalization of P2X7 receptor and Connexin-43 in the spinal cord of EAE rats, specifically in close proximity of CNS-infiltrated CD4+ T cells (Figure 1). This colocalization involves β3-integrin and depends on astroglial mitochondrial activity, and could point to potential new targets for MS therapeutics that modulate these cell-cell interactions.
Figure 1: The P2X7 receptor colocalizes with the astroglial connexin-43 in the immediate proximity of CNS-infiltrated CD4+ T cells. (c) Confocal images of P2X7R (magenta), Cx43 (green), and CD4+ T cell (cyan) immunofluorescent labeling. The depicted regions of interests (ROI) were used for analysis of P2X7R, Cx-43 signal intensity, and the colocalization in the proximity of CD4+ T cells and random ROIs. The yellow and dashed white lines mark the 5 µm and 20 µm radial distances, respectively, and these were measured from the center of the ROIs. Scale bar is 10 µm. (d) Graph showing the distribution of the P2X7R signal intensity in the proximity of CD4+ T cells (magenta) and random ROIs (gray) (two-way ANOVA, p = 0.039). (e) Graph showing the distribution of the Cx-43 signal intensity in the proximity of CD4+ T cells (green) and random ROIs (gray) (two-way ANOVA, p = 0.089).
In another study, a group turned their attention to the role of TRPV4 in mitochondrial function and how this connects to mitochondrial-related diseases (2). Here the researchers used our guinea pig Anti-TRPV4 Antibody (#ACC-034-GP) to examine the colocalization of TRPV4 with mitochondrial proteins MFN1 and MFN2.
In a wonderful series of multiplex assay, the team ran multicolor immunofluorescence and managed to clearly show how endogenous TRPV4 colocalizes and interacts with the endogenous mitochondrial proteins, MFN1 and MFN2 (Figure 2). Their findings prove that TRPV4 interacts with these proteins at endoplasmic reticulum-mitochondrial contact points (MAM), which are crucial for regulating mitochondrial Ca2+ levels and morphology.
Figure 2: TRPV4 colocalises and interacts with MFN1 and MFN2. b. TRPV4 colocalises (indicated by white arrows) with MFN1 or MFN2 in Mesenchymal stem cells isolated from mouse bone marrow. Scale bar: 1 μm. d. TRPV4 colocalises (indicated by white arrows) with MFN1 or MFN2 in mitochondria purified from mouse brain. Distinct colocalisation is observed in some but not in all mitochondria. Scale bar: 1 μm.
This multiplex approach teased out significant interplays that hadn’t been examined before and offered us a new look into how TRPV4 influences mitochondrial dynamics and associated diseases.
Your Flexible Addition
Guinea pig antibodies, while not universally superior to other antibodies, offer you advantages in specific research contexts because they provide options, which means flexibility, in experimental design. Hopefully, it’s clear that these qualities make guinea pig antibodies a valuable addition to lab’s antibody collection.
Take a look at the range and add some options to your research
References
1. K. D. Milicevic, D. B. Bataveljic, J. J. Bogdanovic Pristov, P. R. Andjus, L. M. Nikolic, Astroglial Cell-to-Cell Interaction with Autoreactive Immune Cells in Experimental Autoimmune Encephalomyelitis Involves P2X7 Receptor, β3-Integrin, and Connexin-43. Cells 12, 1786 (2023). DOI: https://doi.org/10.3390/cells12131786.
2. T. K. Acharya, A. Kumar, S. Kumar, C. Goswami, TRPV4 interacts with MFN2 and facilitates endoplasmic reticulum-mitochondrial contact points for Ca2+-buffering. Life Sci 310, 121112 (2022). DOI: https://doi.org/10.1016/j.lfs.2022.121112.
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