Exosomal Protein Characterization

Total proteins of the exosomes were extracted with standard RIPA buffer with PMSF (MCE, United States) at a volume ratio of 99:1, and the concentration of protein was normalized using the BCA assay. The protein samples (25 μg) were then subjected to 10% SDS-PAGE and transferred onto a membrane. The PVDF membrane was incubated with primary antibodies at 4°C overnight, including CD9, CD63, TSG101, and GRP94 (Abcam, ab92726, ab134045, ab125011, and ab238126 Cambs, United Kingdom), and then, the membrane was incubated with HRP-conjugated secondary antibodies (Abcam, Cambs, United Kingdom) for 1 h (Min et al., 2019 (link)).

Free full text: Click here

Xiao Q., Yin R., Wang Y., Yang S., Ma A., Pan X, & Zhu X. (2021). Comprehensive Analysis of Peripheral Exosomal circRNAs in Large Artery Atherosclerotic Stroke. Frontiers in Cell and Developmental Biology, 9, 685741.

Publication 2021

ab92726 ab134045 ab125011 ab238126 HRP-conjugated secondary antibodies

Antibodies Assay Buffer Exosomes Grp94 Membrane Protein Pvdf Ripa Sds page Tsg101

Corresponding Organization :

Other organizations : Qingdao University, Affiliated Hospital of Qingdao University

Top 5 similar protocols

Variable analysis

independent variables

Protein extraction method (RIPA buffer with PMSF)
Protein concentration normalization (BCA assay)

dependent variables

Protein expression levels of CD9, CD63, TSG101, and GRP94

control variables

Protein sample amount (25 μg)
SDS-PAGE (10%)
Membrane type (PVDF)
Primary antibodies (CD9, CD63, TSG101, and GRP94)
Secondary antibodies (HRP-conjugated)

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!