This experiment aims to establish a Gus staining method for detecting the distribution and activity of β-glucuronidase (GUS) in tissues.
Method Article
Gus staining
https://doi.org/10.21203/rs.3.pex-2367/v1
This work is licensed under a CC BY 4.0 License
This protocol has been posted on Protocol Exchange, an open repository of community-contributed protocols sponsored by Nature Portfolio. These protocols are posted directly on the Protocol Exchange by authors and are made freely available to the scientific community for use and comment.
posted
You are reading this latest protocol version
This experiment aims to establish a Gus staining method for detecting the distribution and activity of β-glucuronidase (GUS) in tissues.
Gus staining
β-glucuronidase
tissue sections
substrate staining
Preparation of reagents
(1) GUS staining solution: Take an appropriate amount of DMSO, Na2CO3, and water, mix thoroughly until completely dissolved, add GUS substrate, then add potassium ferrocyanide, and finally add Triton X-100.
(2) Buffer: Prepare an appropriate amount of MES, Na2CO3, and water, mix thoroughly until completely dissolved, and adjust the pH to 5.0.
Tissue processing
(1) Fix the tissue in 10% formaldehyde and store it at 4℃.
(2) Dehydrate the fixed tissue and immerse it in a clearing agent for transparency.
(3) Embed the transparent tissue and make sections.
GUS staining
(1) Place the sections in the GUS staining solution and incubate at 37℃ for 1-2 hours.
(2) Wash with buffer to remove unbound dyes and substrates.
(3) Place the sections in the substrate solution and incubate at 37℃ for 1-2 hours.
(4) Wash with buffer to remove unbound substrate.
Result observation
(1) Observe the staining results under a microscope and record the distribution and activity of GUS.
(2) Take photos or draw images as needed to record the experimental results.
Experimental Summary:
This experiment established a reliable GUS staining method that can be used to detect the distribution and activity of β-glucuronidase in tissues. It is important to follow proper experimental procedures to ensure the accuracy and reliability of the staining results.
This protocol has been posted on Protocol Exchange, an open repository of community-contributed protocols sponsored by Nature Portfolio. These protocols are posted directly on the Protocol Exchange by authors and are made freely available to the scientific community for use and comment.
posted
You are reading this latest protocol version