Improve Research Reproducibility A Bio-protocol resource
Concise Method
tooltip

Immunocytochemistry

IK Imran Khan
SR Saeed Ur Rahman
ET Elieza Tang
KE Karl Engel
BH Bradford Hall
AK Ashok B. Kulkarni
PA Praveen R. Arany
request Request a Protocol
ask Ask a question
Favorite

Immunocytochemistry was performed on collagen gel by first fixing and permeabilized using cold methanol for 5 min at − 20 °C. Following a brief PBS wash, 1% BSA was used to block non-specific staining for 1 h at room temperature. Then, gels were incubated with α-SMA primary antibody overnight (Abcam, 1:100) at 4 °C. The following day, collagen gels were washed in PBS and incubated with a secondary antibody (Anti-rabbit Alexa Fluor 488) for 1 h. Finally, gels were stained with DAPI containing antifade solution and visualized using a confocal scanning electron microscope (Carl-Zeiss).

Copyright and License information: The Author(s) ©2021
Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/.

Do you have any questions about this protocol?

Post your question to gather feedback from the community. We will also invite the authors of this article to respond.

post Post a Question
0 Q&A