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Calcium Assay Service
Overview of Calcium Assay
Introduction of Calcium in Neurons
Calcium (Ca²⁺) is the essential element released from bone to bloodstream in humans. In neuroscience, calcium (Ca²⁺) plays fundamental and diversified roles. As ubiquitous second messenger in multiple signaling pathways, it has been proved that Ca²⁺ participates in multiple biological functions. The cytosolic Ca²⁺ concentration is regulated by the opening of the plasma membrane and subcellular Ca²⁺ sensitive channels and ultimately leads to the flexibility of Ca²⁺ signaling. The endoplasmic reticulum (ER) in the cell can accumulate the concentration of Ca²⁺ to 10-100 mM, and the release of Ca²⁺ is mediated by RyRs and IP3Rs. In addition, mitochondria can also store Ca²⁺ and play an important role in Ca²⁺ transients. Due to the central role in neuronal physiology, disruption of Ca²⁺ signaling and homeostasis would result in neurodegenerative disorders, as well as aging. There are two types of plasma membrane Ca²⁺ channels for the regulation of neuronal Ca²⁺ influx, including voltage-dependent Ca²⁺ channels (VDCCs) and receptor-operated (ligand-gated) channels (ROCs).
Fig.1 Cav channel subunit interactions.2
Functions of Calcium
- Regulation of neuronal gene expression
- Energy production
- Membrane excitability
- Synaptogenesis
- Synaptic transmission
- Development of dendrite
- Memory storage and information process
Fig.2 Some of the pathways modulating Cav2 channel function.2
Our Calcium Assay Service
Creative Biolabs provides an easy and quantitative method for the measurement of calcium in different biological fluids. Firstly, N,N,N',N'-tetrakis-(2-pyridylmethyl)ethylenediamine (TPEN) would be used to avoid the influence of other divalent metals, like Zine. Secondary, the proprietary probe can specifically bind to calcium cations and high fluorescence signal can then be generated. Finally, the concentration of calcium can be quantified using a microplate reader as the fluorescence is directly proportional to the amount of calcium.
Features of Our Service
- Simple operation process
- Rapid detection (within 10 minutes)
- High sensitivity
- Wide range of applications
- Multiple sample types
Creative Biolabs is one of the well-recognized experts who are professional in applying advanced platforms for a broad range of neurosciences research, now we provide the novel calcium assay service for our clients all over the world. We are pleased to use our extensive experience to offer the best service and the most qualified products to satisfy each demand from our customers. If you are interested in our services and products, please do not hesitate to contact us for more detailed information.
Published Data
Microglial calcium signaling underlies many key physiological and pathological processes. Anthony D Umpierre et al. assessed how microglial calcium signaling responds to changes in neuronal activity across a broad spectrum using multiple GCaMP6 variants targeting microglia.
They studied spontaneous calcium activity in awake, head-restrained animals using two-photon microscopy to assess microglia in layers I and II/III of somatosensory cortex. Microglial calcium activity was present in both Lck-GCaMP6f mice and cytoplasmic GCaMP6s mice. However, the higher fluorescence amplitude and slower decay kinetics of GCaMP6s animals make this fluorophore ideal for studying calcium activity in thin microglia protrusions.
Combined with data from other studies, the results of this study suggest that microglia protrusion calcium signaling is compatible with changes in neuronal activity, and that both hypo- and hyperactivity of neurons trigger an increase in microglia protrusion calcium signaling.
Fig. 3 Spontaneous microglial calcium activity reported in the Lck-GCaMP6f and cytosolic GCaMP6s mouse.3
Applications
- Neuronal Activity Monitor. Neuronal activity leads to the influx of Ca2+ through voltage-gated calcium channels. Calcium assays allow researchers to monitor these changes in intracellular calcium levels, providing insights into neuronal firing and synaptic activity.
- Studying Synaptic Plasticity. Changes in synaptic strength, key to learning and memory, are associated with Ca2+ signaling. Calcium assays help in studying these processes by allowing the measurement of calcium transients during synaptic modification.
- Drug Screening and Pharmacological Research. Calcium assays are used to assess how different pharmacological agents affect calcium signaling pathways. This is valuable in identifying compounds that modulate neuronal activity and could be potential treatments for neurological disorders. Automated calcium imaging systems enable the screening of large libraries of compounds to identify those that affect neuronal calcium signaling.
- Understanding Neurotransmitter Release. Calcium influx is a trigger for the release of neurotransmitters from synaptic vesicles. Calcium assays help in studying the mechanisms of neurotransmitter release and the role of different calcium channels and sensors in this process.
- Investigating Neurodegenerative Diseases. Abnormalities in calcium homeostasis are linked to various neurodegenerative diseases such as Alzheimer's, Parkinson's, and Huntington's disease. Calcium assays aid in understanding how these disturbances contribute to disease progression.
FAQs
Q: What types of samples can be analyzed with your calcium assay service?
A: Our calcium assay service can handle a variety of sample types, including cultured neurons, brain slices, and isolated cells from different regions of the nervous system. We can also analyze samples from animal models or human-derived cell lines. If you have a specific type of sample, please contact us to discuss its compatibility and any special handling requirements.
Q: What is the sensitivity and detection limit of your calcium assay?
A: Our calcium assay employs highly sensitive fluorescent and luminescent indicators that can detect even small changes in intracellular calcium levels. The exact sensitivity and detection limits depend on the specific indicator used and the experimental setup. Generally, we can detect calcium concentrations in the nanomolar to micromolar range, allowing us to capture subtle changes in calcium dynamics. For precise details regarding sensitivity for your specific application, please contact us.
Q: Can you work with specific calcium indicators that we provide?
A: Yes, we can work with specific calcium indicators that you provide, provided they are compatible with our equipment and protocols. If you have a preferred indicator, we will evaluate its suitability for your assay and integrate it into our workflow. Our team will collaborate with you to ensure the optimal setup for your experiments, leveraging your preferred reagents while maintaining high-quality standards.
Q: Do you offer technical support after the assay results are delivered?
A: Yes, we offer comprehensive technical support after delivering the assay results. Our team of experts is available to help you interpret the data, answer any questions, and provide guidance on further experiments or analyses. We aim to support your research journey and ensure you can make the most of the results from our calcium assay service. Feel free to reach out to us with any follow-up inquiries.
Scientific Resources
References
- Kawamoto, E.; et al. Physiology and pathology of calcium signaling in the brain. Frontiers in pharmacology. 2012, 3: 61.
- Dolphin, A.; Lee, A. Presynaptic calcium channels: specialized control of synaptic neurotransmitter release. Nature Reviews Neuroscience. 2020, 21(4): 213-229.
- Umpierre, Anthony D., et al. "Microglial calcium signaling is attuned to neuronal activity in awake mice." Elife 9 (2020): e56502.
For Research Use Only. Not For Clinical Use.