Immunohistochemical Analysis of Soleus Muscle

Serial (10 μm) transverse sections of the soleus mid-belly region were taken on a CM1850 cryostat (Leica Biosystems, Buffalo Grove, IL) and mounted on gelatin-coated glass slides. Immunohistochemistry (n = 6-8/group) was performed for determination of fiber cross-sectional area (fCSA) and fiber type distribution, as previously reported [25 (link),30 (link),31 (link)]. Antibodies used were as follows: rabbit anti-laminin (1:200; Thermo Scientific, Waltham, MA), rhodamine-conjugated goat anti-rabbit IgG (1:500), Alexa Fluor® 488 goat anti-mouse IgG (1:200), Alexa Fluor® 488 goat anti-mouse IgM (1:200) all from Life Technologies (Carlsbad, CA); and anti-MHC I (BA.D5), anti-MHC IIA (SC.71), anti-MHC IIB (BF.F3), and anti-MHC IIX (BF.35) obtained from the Developmental Studies Hybridoma Bank (Iowa City, IA). Image acquisition was performed on an Eclipse TE2000-S microscope (Nikon Instruments, Melville, NY) with the fiber type distribution and average fCSA determined, in a blinded manner by consensus of two investigators, from an analysis of 300–350 total fibers per animals with Image J (NIH).

Free full text: Click here

Phillips E.G., Beggs L.A., Ye F., Conover C.F., Beck D.T., Otzel D.M., Ghosh P., Bassit A.C., Borst S.E, & Yarrow J.F. (2018). Effects of pharmacologic sclerostin inhibition or testosterone administration on soleus muscle atrophy in rodents after spinal cord injury. PLoS ONE, 13(3), e0194440.

Publication 2018

CM1850 cryostat rabbit anti-laminin Alexa Fluor® 488 goat anti-mouse IgG Alexa Fluor® 488 goat anti-mouse IgM Eclipse TE2000-S microscope

Alexa fluor 488 Animals Anti igg Anti igm Antibodies Buffalo Fiber Gelatin Goat Hybridoma Immunohistochemistry Laminin Microscope Mouse Rabbit Rhodamine Soleus

Corresponding Organization : Florida College

Top 5 similar protocols

Protocol cited in 1 other protocol

Variable analysis

independent variables

Experimental groups

dependent variables

Fiber cross-sectional area (fCSA)
Fiber type distribution

control variables

Transverse sections of the soleus mid-belly region
Gelatin-coated glass slides
Antibodies used for immunohistochemistry
Total fibers analyzed per animal (300-350)
Blinded analysis by two investigators

controls

Positive control: Not explicitly mentioned
Negative control: 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!