Immunocytochemistry and Immunoblot Assays for Myotubes

For immunocytochemistry assays, myotubes were fixed in cold methanol (−20 °C) for 30 min and permeabilized with 0.05% Tween 20 in PBS for 1 min, as previously described [7 (link),8 (link),16 (link),17 (link)]. For immunoblot assays, solubilized lysate of myotubes (15 μg of total protein) was subjected to SDS–PAGE (8 or 10% gel), as previously described [7 (link),8 (link),16 (link),17 (link),18 (link),19 (link),20 (link)]. Anti-RyR1 and anti-SERCA1a antibodies were obtained from Affinity BioReagents. Anti-DHPR, anti-Orai1, anti-STIM1, and anti-α-actin antibodies were obtained from Abcam (Cambridge, MA, USA).

Free full text: Click here

Park J.H., Jeong S.Y., Choi J.H, & Lee E.H. (2021). Pathological Mechanism of a Constitutively Active Form of Stromal Interaction Molecule 1 in Skeletal Muscle. Biomolecules, 11(8), 1064.

Publication 2021

Anti-DHPR anti-Orai1 anti-STIM1 anti-α-actin

Actin Anti antibodies Assays Cold Immunoblot assays Immunocytochemistry Methanol Myotubes Protein Ryr1 Sds page Stim1 Tween 20

Corresponding Organization : Catholic University of Korea

Top 5 similar protocols

Variable analysis

independent variables

Methanol fixation temperature (-20 °C)
Methanol fixation duration (30 min)
Permeabilization with 0.05% Tween 20 in PBS (duration not specified)

dependent variables

Immunocytochemistry assays on myotubes
Immunoblot assays on solubilized lysate of myotubes (15 μg of total protein)

control variables

Antibodies used for immunocytochemistry and immunoblot assays (Anti-RyR1, Anti-SERCA1a, Anti-DHPR, Anti-Orai1, Anti-STIM1, Anti-α-actin)

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!