As a leader in the complement field, Creative Biolabs is committed to the development of future biosciences, contributing high-quality complement products and leading analytical technologies for comprehensive and customized solutions. With our highly specialized technical capabilities and extensive experience in the field of complement, we are well prepared and capable of facilitating collaborations in the fast-growing life science research field. Our experienced scientists will work with you to customize a sensible solution to advance your complement program.
Detection Protocol of C5a-Mediated Increase in Intracellular Calcium
The crucial role of C5a in infection, allergy, autoimmune disease and cancer reveals that it is a key regulator of pro-inflammatory and immunomodulatory effects. C5aR exerts most of its biological functions by activating the specific receptor C5aR. C5aR activation involves multiple signaling pathways, ultimately leading to increased intracellular calcium concentration. Here, we describe a simple flow cytometry-based protocol for the detection of C5a-mediated increase in intracellular calcium.
Fig.1 Flow chart of detection protocol of C5a-mediated increase in intracellular calcium. (Creative Biolabs)
In recent years, with the continuous deepening of biotechnology research, the demand for customized complement services and corresponding products has expanded rapidly. With a wealth of experience and a professional team, Creative Biolabs has gradually become a world-leading complement supplier. We are committed to providing strictly tested complement products and one-stop customized services for global customers.
Published Data
Fig.2 Calcium transient analysis reveals interactive signaling elicited by concurrent C3a and C5a activation.1
To evaluate whether C3a and C5a induce alterations in the RPE, researchers employed cultured RPE cells, assessing singular and combined stimuli via fluorescence microscopy, western blotting, and immunohistochemistry. Calcium transient analysis revealed interactive signaling during C3a and C5a stimulation, with the temporal profiles of Ca2+ responses, including both peak and late phases, meticulously examined in ARPE-19 cells under varied conditions. Complement activation triggered increased cytosolic Ca2+, PI3-kinase/Akt engagement, and phosphorylation of FoxP3 and FOXO1, accompanied by cytokine and chemokine release. Differential outcomes between simultaneous and sequential anaphylatoxin exposure underscore that coordinated C3aR/C5aR signaling finely tunes RPE immunoregulatory dynamics, thereby enhancing our understanding of AMD pathophysiology.
Services
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Complement function/activity test
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Complement activation products test
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Complement inhibitor validation
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Complement genetic test
Why Choose Us?
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Professional team - our complement detection platform is equipped with a top-notch technical team, who are specialized in complement detection.
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Advanced platform - our complement platform is equipped with companion instruments, which allows us to perform a variety of assays for complement detection, such as flow cytometry and immunofluorescence.
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Reliable results - standardized complement assays provide the foundation for reliable experimental results.
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Customized solution - complete complement detection solutions can be customized according to unique needs.
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Please note that our protocols are only for your reference!
Reference
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Busch, Catharina, et al. "Anaphylatoxins activate Ca2+, Akt/PI3-kinase, and FOXO1/FoxP3 in the retinal pigment epithelium." Frontiers in Immunology 8 (2017): 703. Distributed under Open Access license CC BY 4.0, without modification.
For Research Use Only.
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