Localization Service of Lysosomal Ion Channels

Background of Lysosomal Ion Channels

Lysosomal ion channels are emerging as essential proteins for intracellular homeostasis and as pharmacological targets for many human pathologies. In response to environmental and cellular signals, lysosomes change their location, number, morphology, size, composition, and activity over short periods of time to meet changing cellular demands. Ion channels in lysosomes are important transduction factors mediating signal-initiated Ca²⁺/Fe²⁺/Zn²⁺ release and H⁺/Na⁺/K⁺-dependent changes in membrane potential. Misregulation of lysosomal ion flux impairs lysosomal motility, membrane transport, nutrient sensing, membrane repair, organelle membrane contact, and lysosomal biogenesis and adaptation. To investigate how cellular signaling regulates lysosomal ion flux and membrane potential changes in intact cells, the first task in identifying lysosomal channels is to determine their intracellular localization.

Fig. 1. Monitoring lysosomal Ca²⁺ release using genetically-encoded Ca²⁺ indicators and Fura-2. (Zhong XZ, et al., 2017)

Lysosomal Ion Channels Localization Services

Fluorescent proteins fused to the target proteins provide a useful tool to virtualize protein localization in live cells. We have provided multiple strategies to localize lysosomal proteins. Here at CD BioSciences, we are committed to providing expert lysosomal ion channel localization services to our clients worldwide. Our biologists use immunostaining target channels to monitor the subcellular localization of endogenous lysosomal channels without interference from exogenous overexpression. Currently we can localize the following lysosomal ion channels.

• Lysosomal H⁺ channels
• Lysosomal Ca²⁺ channels
• Lysosomal Na⁺ channels
• Lysosomal K⁺ channels
• Lysosomal Fe²⁺ channels
• Lysosomal Zn²⁺ channels
• Lysosomal Cl^- channels

We fuse co-expression target channels and LAMP1 with different fluorescent proteins such as GFP (green), mCherry/RFP/DsRed (red) or YFP (yellow), or label channel-expressing cells with dextran. We have successfully performed intracellular localization of the following lysosomal ion channels.

• Endogenous P2X4 was detected in lysosomes by immunofluorescence staining.
• Detection of lysosomal ion channel proteins in lysosome-associated membrane protein 1 (Lamp1) fractions by immunoblotting with the help of specific antibodies.

Why Choose Us

• Immunostaining was used to detect lysosomal ion channel protein localization without interference from exogenous overexpression.
•  Widely used to validate the application of fluorescent fusion proteins in heterologous systems.
•  Widely used for immunostaining of endogenous proteins to study the subcellular localization of lysosomal channels.
•  Experienced team.
•  Standard scientific customized services.

CD BioSciences can meet any reasonable needs of our clients, taking time and budget into consideration for you. Our aim is to be customer-centric and to provide the highest quality services to customers. Our customer service representatives are enthusiastic and trustworthy 24 hours a day, 7 days a week. If you are interested in our services, please feel free to contact us for more information or a detailed discussion.

Reference

1. Zhong XZ, Yang Y, et al. (2017) Methods for monitoring Ca²⁺ and ion channels in the lysosome. Cell Calcium. 64:20-28.