The Power of Product Combinations in Your Research

Venom-derived peptide toxins for Ca_v channels, a primary antibody for TRPV1, and a selective blocker of Na_v channels; Having a reliable spectrum of reagents, lets you tackle complex questions with confidence.

We’re all guilty of sticking with what we know: whether it’s the same protocols handed down from scientist to scientist in the lab, or the same vial from the same vendor that’s been in the same fridge for months. More often than not it comes from simply never having had the time to really look around at what else is available, or just not wanting to risk making a change. Biology can be messy and when things work, we don’t like to change it too much – especially with so much variability in most reagents.

But what if you could get a whole range of reagents from a single vendor who has been making everything in house for decades? From antibodies to toxins to small active molecules, all made by the same group of scientists and held to the same rigorous QC standards. That’s what we do at Alomone Labs. We’re ion channels and membrane protein specialists who make and test everything right here in our labs and then send it straight to you.

Here’s a great demonstration of the power of getting reliable multiple reagents in one place. You’ll see the researchers in this new paper explore complex mechanisms of neuropathic pain with products from different categories to get comprehensive and insightful results. Let’s take a look.

Getting to Grips with Neuropathic Pain

Neuropathic pain comes from abnormal sensory processing due to nerve damage. In a recent study, a team from the University of Calgary focused on the involvement of voltage-gated calcium channels (Ca_V2.1 and CaV2.2) in the increased calcium transients in TRPV1 DRG neurons, under neuropathic pain conditions (1).

Focusing on Pain Neurons with Anti-TRPV1 Antibodies

To demonstrate calcium transients in TRPV1 DRG neurons, the researchers combined genetic manipulations with 2-photon calcium imaging. The genetically encoded calcium reporter GCaMP6s was engineered to be expressed in TRPV1 nociceptors by crossing a TRPV1-Cre mouse line with a cre-dependent GCaMP6s mouse line.

The expression profile of the calcium reporter GCaMP6a and TRPV1 channels in superficial layers of the spinal dorsal horn was demonstrated using Anti-GFP Antibody and Anti-TRPV1 (VR1) Antibody (#ACC-030), respectively (Figure 1).

 

 _{Figure 1. Confocal images of GCaMP6s and Trpv1 expression in superficial layers of the dorsal horn of a Trpv1-Cre x Ai96 mouse (L4–L5). Lumbar sections of the spinal cord (L4 to L5) were immunostained with GFP Ab (stains GCaMP6s—green) and Trpv1 Ab (magenta). The merged image indicates GCaMP6s and Trpv1 co-expression in superficial layers of the dorsal horn (bottom panel). Scale bar: 50 μm. Modified from Ferron et al. (2024) (1)}

Venom-Derived Toxin Inhibitors: Conotoxin and Agatoxin

Next, the researchers used ω-Conotoxin GVIA (#C-300) and ω-Agatoxin IVA (#STA-500) – both synthetic and biologically active peptide toxins – to probe the contribution of Ca_V2.1 and Ca_V2.2 channels to the enhanced calcium transients. These toxins block calcium channels with high specificity and potency. Through clever use of these toxins, they were able to demonstrate the dysregulation of N- and P/Q-type channels in DRG sensory neuron nerve terminals after peripheral nerve injury, specifically at the presynaptic terminals. Blocking these channels with ω-conotoxin GVIA and ω-agatoxin IVA showed a substantial decrease in calcium transients in a neuropathic pain model. This provided clear evidence of their involvement in the pathological state (Figure 2).

_{Figure 2. (A and B) GCaMP6s fluorescence variations from Trpv1+ DRG neuron central terminals in superficial layers of the spinal dorsal horn in response to 1 pulse (A) and 10 pulses (B) at 3 mA before (Ctrl), 20 min after application of ω-conotoxin GVIA (GVIA, 1 μM) and 20 min after adding ω-agatoxin IVA (Aga, 200 nM, Aga+GVIA) to GVIA. (G) Variations of N- and P/Q-type Ca2+ channel protein expression in ipsilateral L4 DRG following SNI treatment. ELISA techniques were used to quantify total channel protein concentration from DRG. Proteins were extracted from individual ipsilateral L4 DRGs (5 DRGs for each condition). Because of low extraction yield, 2 samples had to be pooled to obtain enough total protein to reliably run the ELISA. N-type total protein. Scale bar: 50 μm. All error bars reflect S.E.M. Modified from Ferron et al. (2024) (1)}

Untangling Neuronal Activity with Tetrodotoxin

Finally, we see the researchers put Tetrodotoxin citrate (#T-550) (TTX), a natural classic blocker of voltage-gated sodium (Na_V) channels, to untangle the dependence of calcium transients on neuronal activity. TTX then helped distinguish between calcium influx due to neuronal firing and other causes. With TTX, the researchers saw calcium transients totally abolished, revealing the critical role of neuronal activity in the phenomenon of calcium transients under neuropathic conditions. This finding is essential for understanding how neuronal activity contributes to pain signaling. You might want to consider using TTX if you need to differentiate sources of calcium influx in your own studies.

Benefits of Using Alomone’s Comprehensive Product Range

While some vendors also work like resellers and simply stock reagents from as many biological disciplines as possible, we’re focused. We’re ion channels and other membrane protein specialists. And we make everything right here at Alomone Labs. That means you get:

1. A specialized selection: We work hard to make sure that whatever your ion channels and membrane protein research needs, we’ve made it for you. This specialized selection gives you access to venom-derived peptide toxins, like agatoxin and conotoxin, and biologically active compounds such as TTX, but also to antibodies, blocking peptides, growth factors and neurotrophins. 
2. Flexibility and customization: As the scientists who both make and test all of our products, we can easily generate customized reagents, like fluorescently labeled primary antibodies, as well as venom-derived peptide toxins, to better suit your research. If something isn’t available off the shelf, we have dedicated Advanced Primary Antibody Conjugation Services so you can always get your reagents your way – no compromises needed.
3. Reliability: We have complete oversight of the reagent development process from design to QC. That means nothing goes unnoticed and you benefit from reliable, consistent products. Also, we keep a steady level of stock across our range of ion channels and membrane protein products. So, while some vendors may struggle to fulfill your order, we have plenty available and you can choose from multiple vial sizes. All you have to do is pick which combination of items you want and leave it to us to send it out – with free international shipping.
4. Value: When you buy more, you save more. We offer a rewards plan like this: with our Reagent Rewards you can choose a bigger selection of reagents and save money at the same time. The rewards plan lets you earn up to seven free, full-sized products, depending on how much you choose. Don’t ever let your scientific curiosity be held back by budget.

Add Alomone Labs Products to Your Research

The neuropathic pain study here shows the value of using multiple Alomone Labs products together. Combining antibodies, toxins, and small molecules helped uncover intricate details that single-product studies might miss, which enhances the depth of research and opens new avenues for understanding.

For example, the study demonstrated that blocking both N- and P/Q-type calcium channels significantly reduced pain-associated calcium transients. The use of anti-TRPV1 antibody allowed precise identification of pain-transmitting neurons, confirming the specificity of the studied mechanism. TTX helped distinguish the role of neuronal activity in pain signaling, underscoring the importance of sodium channels in this process.

Already using one of these products? Try adding a broader selection of Alomone Labs reagents to your assays – it could save you time and money, and even deliver more reliable results. Whether you study ion channels, pain pathways or neuronal activity, our reliable products give you the necessary tools.

Alomone Labs products mentioned in this blog:

Reference

1.      L. Ferron, E. K. Harding, M. A. Gandini, C. Brideau, P. K. Stys, G. W. Zamponi, Functional remodeling of presynaptic voltage-gated calcium channels in superficial layers of the dorsal horn during neuropathic pain. iScience 27, 109973 (2024).