DRGs (L1–L6) were isolated from Vglut2-ChR2-YFP mice and cultured in poly-L lysine (100 µg/mL) and laminin (50 µg/mL) precoated chamber plates for 48 h. Cells were stimulated using 470 nm LED light (or control yellow light at 595 nm) at 20 Hz, and 10% duty cycle for 30 min using fiber-coupled LEDs (Thorlabs Inc). The supernatant was collected over time (for HMGB1 release) or for an additional 60-min incubation for measurements of LDH release (Sigma-Aldrich). For HMGB1 staining, cultured DRG cells were fixed with 4% paraformaldehyde (Thermo Fisher Scientific) followed by immunofluorescent staining with anti-HMGB1 antibody Alexa-555 (red, Abcam) and DAPI (blue) with endogenous VGlut2-YFP. Images were acquired using an LSM880 laser scanning confocal microscope (Zeiss). Levels of secreted HMGB1 in the supernatant were quantitated using an ELISA kit (IBL International).
Ldh release
Manufactured by Merck Group
The LDH release assay is a laboratory tool used to measure the activity of the enzyme lactate dehydrogenase (LDH) released from damaged or disrupted cells. It provides a quantitative assessment of cellular cytotoxicity or cell membrane integrity. The assay is commonly used in various in vitro cell-based experiments and research applications.
Automatically generated - may contain errors
2 protocols using ldh release
1
Optogenetic Stimulation of DRG Neurons
2
Quantifying Cell Cytotoxicity via LDH Assay
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
LDH release assays (Sigma #4744926001) were performed as previously described (31) . Cell media supernatants from experimental groups were collected and run in biological triplicate and technical triplicate. Basal cell media served as a background control and cell media harvested from a lysis group served as a positive control for 100% LDH release. Plates were read on a CLARIOstar plate reader per manufacturer's instruction. Raw data were first analyzed in excel, then statistical analyses and graphing were performed using GraphPad Prism software.
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!