Immunohistochemical Analysis of Tibialis Anterior

Tibialis anterior muscles (TA) were fixed in 10% formalin for 24 h and embedded in paraffin. Immunohistochemistry was carried out as previously described (2 (link)) using antibodies directed against cluster of differentiation 68 (CD68), 4-hydroxy-2-nonenal (HNE), interleukin 1-beta detecting mature and precursor form (IL-1β), interleukin 18 detecting only mature form (IL-18), peroxiredoxin 3 (PRDX3), and tumor necrosis factor alpha (TNFα) (Supplementary Information, Table S1). Whole muscle sections were scanned, and then the optical density of diaminobenzidine (DAB, ThermoFisher-Scientific, Waltham, MA, USA) deposits (IL‐1β, TNFα, HNE and PRDX3) or the percentage of stained areas (IL-18, CD68) was quantified using Fiji (NIH, Maryland, USA). TA sections were also stained with haematoxylin and eosin. In addition, Quadriceps (Q) sections (described below) were stained with Picro-Sirius red (Abcam, Cambridge, UK) to evaluate muscle fibrosis. Fibrotic tissue was scored setting a color balance threshold and data obtained were expressed as the percentage of total section area.

Free full text: Click here

Dubuisson N., Davis-López de Carrizosa M.A., Versele R., Selvais C.M., Noel L., Van den Bergh P.Y., Brichard S.M, & Abou-Samra M. (2022). Inhibiting the inflammasome with MCC950 counteracts muscle pyroptosis and improves Duchenne muscular dystrophy. Frontiers in Immunology, 13, 1049076.

Publication 2022

diaminobenzidine (DAB, Picro-Sirius red

4 hydroxy 2 nonenal Antibodies Embedded paraffin Eosin Fibrotic Formalin Il 18 Il 1β Immunohistochemistry Interleukin 18 Muscle Optical Peroxiredoxin 3 Quadriceps Tibialis anterior muscles Tissue Tnfα

Corresponding Organization : Cliniques Universitaires Saint-Luc

Other organizations : Universidad de Sevilla

Top 5 similar protocols

Protocol cited in 1 other protocol

Variable analysis

independent variables

Antibodies directed against cluster of differentiation 68 (CD68)
Antibodies directed against 4-hydroxy-2-nonenal (HNE)
Antibodies directed against interleukin 1-beta detecting mature and precursor form (IL-1β)
Antibodies directed against interleukin 18 detecting only mature form (IL-18)
Antibodies directed against peroxiredoxin 3 (PRDX3)
Antibodies directed against tumor necrosis factor alpha (TNFα)

dependent variables

Optical density of diaminobenzidine (DAB) deposits (IL‐1β, TNFα, HNE and PRDX3)
Percentage of stained areas (IL-18, CD68)
Muscle fibrosis (percentage of total section area)

control variables

Tibialis anterior (TA) muscles fixed in 10% formalin for 24 h and embedded in paraffin
Quadriceps (Q) sections stained with Picro-Sirius red

positive controls

Not explicitly mentioned

negative controls

Not explicitly mentioned

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!