Immunofluorescence of Tissue-Derived Cells

For immunofluorescence studies, we cultured cells extracted from full-thickness samples12 (link). The cells were grown on cover slips with Rat Tail Collagen I (Gibco, Invitrogen) coating, and fixed with methanol for 5 min at −20 °C⁵⁶. The cover slip samples were incubated one hour at room temperature with the MTCO2 (Abcam, ab3298) and CARD6 antibodies. Alexa Fluor 555 and 488 conjugated IgGs (Invitrogen, Molecular Probes) were used as secondary antibodies and the nuclei stained with DAPI (Sigma-Aldrich). The pictures were taken with Zeiss LSM 5 Duo confocal microscope.

Free full text: Click here

Tervaniemi M.H., Katayama S., Skoog T., Siitonen H.A., Vuola J., Nuutila K., Sormunen R., Johnsson A., Linnarsson S., Suomela S., Kankuri E., Kere J, & Elomaa O. (2016). NOD-like receptor signaling and inflammasome-related pathways are highlighted in psoriatic epidermis. Scientific Reports, 6, 22745.

Publication 2016

Rat Tail Collagen I ab3298 Alexa Fluor 555 and 488 conjugated IgGs DAPI LSM 5 Duo confocal microscope

Alexa fluor 555 Antibodies Cells Collagen i Confocal microscope Cultured cells Dapi Immunofluorescence Methanol Molecular probes Nuclei Tail

Corresponding Organization :

Other organizations : Folkhälsans Forskningscentrum, Karolinska Institutet, University of Helsinki, Helsinki University Hospital, University of Oulu, Science for Life Laboratory

Top 5 similar protocols

Variable analysis

independent variables

Culture conditions (cells extracted from full-thickness samples)
Substrate coating (Rat Tail Collagen I)
Fixation method (methanol for 5 min at −20 °C)
Primary antibodies (MTCO2 and CARD6)

dependent variables

Localization and expression of MTCO2 and CARD6 proteins

control variables

Incubation time (1 hour at room temperature)
Secondary antibodies (Alexa Fluor 555 and 488 conjugated IgGs)
Nuclear stain (DAPI)
Imaging method (Zeiss LSM 5 Duo confocal microscope)

Annotations

Based on most similar protocols

Etiam vel ipsum. Morbi facilisis vestibulum nisl. Praesent cursus laoreet felis. Integer adipiscing pretium orci. Nulla facilisi. Quisque posuere bibendum purus. Nulla quam mauris, cursus eget, convallis ac, molestie non, enim. Aliquam congue. Quisque sagittis nonummy sapien. Proin molestie sem vitae urna. Maecenas lorem.

As authors may omit details in methods from publication, our AI will look for missing critical information across the 5 most similar protocols.

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?

Revolutionizing how scientists
search and build protocols!