Protocol detail

Aggresome Detection in HeLa Cells

Find Similar Protocols

WT or PACSIN1 KO HeLa cells were treated with 20 μM MG132 or 2.5 μg/mL puromycin for 6 h. Detergent soluble-insoluble proteins were fractionated using 1% triton-X 100 with PBS [62 (link)]. After centrifugation, the supernatant was used as the soluble fraction. The pellet was extracted with RIPA buffer included 7 M Urea and used as the insoluble fraction. Aggresome staining was performed by aggresome detection kit (abcam, ab139486) according to the manufacturer’s instruction. The number of aggresome dots normalized per cell were quantified by Fiji.

Free full text: Click here

Oe Y., Kakuda K., Yoshimura S.I., Hara N., Hasegawa J., Terawaki S., Kimura Y., Ikenaka K., Suetsugu S., Mochizuki H., Yoshimori T, & Nakamura S. (2022). PACSIN1 is indispensable for amphisome-lysosome fusion during basal autophagy and subsets of selective autophagy. PLoS Genetics, 18(6), e1010264.

Publication 2022

aggresome detection kit

Buffer Cell Centrifugation Detergent Hela cells Mg132 Proteins Puromycin Ripa Triton x 100 Urea

Top 5 similar protocols

Variable analysis

independent variables

Treatment with 20 μM MG132
Treatment with 2.5 μg/mL puromycin

dependent variables

Number of aggresome dots normalized per cell

control variables

WT (wild-type) or PACSIN1 KO (knockout) HeLa cells
Detergent soluble-insoluble protein fractionation using 1% triton-X 100 with PBS
Aggresome staining using aggresome detection kit (abcam, ab139486)

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!