Western Blotting of Platelet Proteins

Western blotting was performed by modifying a previously described procedure [26 (link)]. Briefly, platelets were lysed in buffer A and B and centrifuged at 20,000× g for 30 min, and the supernatant was collected to obtain the cell membrane lysate. Then, proteins were separated using sodium dodecyl sulfate polyacrylamide gel electrophoresis and reacted with anti-PKC-α antibody (1:500, Santa Cruz), anti-PKC-βI (1:500, Santa Cruz), anti-PKC-βII (1:500, Santa Cruz), anti-PKC-γ (1:500, Santa Cruz), anti-PKC-δ (1:500, Santa Cruz), or anti-actin (1:3000, Cell Signaling) overnight. All samples were analyzed using a LAS 4000 mini (Fuji Photo Film, Tokyo, Japan). This experiment was repeated a total of three times.

Free full text: Click here

Lee B.K., Jee H.J, & Jung Y.S. (2021). Aβ1–40-Induced Platelet Adhesion Is Ameliorated by Rosmarinic Acid through Inhibition of NADPH Oxidase/PKC-δ/Integrin αIIbβ3 Signaling. Antioxidants, 10(11), 1671.

Publication 2021

anti-PKC-βII anti-PKC-γ anti-PKC-δ anti-actin LAS 4000 mini

Actin Anti antibody Buffer Cell membrane Pkc α Pkc βii Pkc γ Platelets Proteins Sodium dodecyl sulfate polyacrylamide gel electrophoresis

Corresponding Organization : Ajou University

Top 5 similar protocols

Variable analysis

independent variables

Modification of previously described procedure [26]

dependent variables

Protein expression levels of PKC-α, PKC-βI, PKC-βII, PKC-γ, PKC-δ, and actin

control variables

Centrifugation speed (20,000× g)
Centrifugation duration (30 min)
Protein separation using sodium dodecyl sulfate polyacrylamide gel electrophoresis
Antibody dilutions (anti-PKC-α 1:500, anti-PKC-βI 1:500, anti-PKC-βII 1:500, anti-PKC-γ 1:500, anti-PKC-δ 1:500, anti-actin 1:3000)
Overnight incubation of samples with antibodies
Analysis using LAS 4000 mini

controls

No positive or negative controls were 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!