To remove the follicle cell layer, Xenopus laevis oocytes were treated with 75 μg/ml Liberase TM research-grade enzyme (Sigma-Aldrich) for 40–60 min in Ca2+-free ND96 solution (96 mM NaCl, 2 mM KCl, 1 mM MgCl2, and 5 mM HEPES, pH adjusted to 7.4 with NaOH). Stage V-VI oocytes were injected with cRNA encoding proteins of interest and incubated for 1–5 d at 16 °C in storage solution before recording. Storage solution contained 96 mM NaCl, 2 mM KCl, 1.8 mM CaCl2, 1 mM MgCl2, 1 mM HEPES, and 10 mg/L gentamycin, with pH adjusted to 7.4 with NaOH. For experiments with dEAG channels co-expressed with CaMKII, cRNA was pre-mixed and injected into the oocytes. To apply CaMKII inhibitory peptide, diluted inhibitory peptide was injected into oocytes expressing dEAG channels 2–4 hours before recording. The final concentration of diluted inhibitory peptide in the oocytes is estimated to be 80 μM.
Liberase research grade enzyme
Manufactured by Merck Group
Liberase TM is a research-grade enzyme solution designed for tissue dissociation and cell isolation. It is a blend of purified enzymes that effectively break down the extracellular matrix, allowing for the release of individual cells from tissue samples. The core function of Liberase TM is to facilitate the extraction of viable cells from various tissue types for further analysis and experimentation.
Automatically generated - may contain errors
Lab products found in correlation
2 protocols using liberase research grade enzyme
1
Xenopus Oocyte Follicle Cell Removal
2
Xenopus Oocyte Follicle Cell Removal
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
To remove the follicle cell layer, Xenopus laevis oocytes were first isolated with forceps and then treated with 75 µg/ml Liberase TM research-grade enzyme (Sigma-Aldrich) for 40–60 min in Ca2+-free ND96 solution that contained 96 mM NaCl, 2 mM KCl, 1 mM MgCl2, and 5 mM HEPES, with pH adjusted to 7.4 with NaOH. Stage IV and V oocytes were injected with WT or mutant cRNA and incubated for 1–5 d at 16°C in storage solution before voltage-clamp experiments. Storage solution contained 96 mM NaCl, 2 mM KCl, 1.8 mM CaCl2, 1 mM MgCl2, 1 mM HEPES, and 10 mg/L gentamycin, with pH adjusted to 7.4 with NaOH.
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!