Western Blotting of METTL14, CSNK1D, and SLC35E1

Western blotting was conducted as previously described (22 (link)). Briefly, protein concentrations were measured with a BCA Kit. Protein lysates were resolved using SDS–PAGE and transferred onto PVDF membranes (Millipore). The membrane was subsequently incubated overnight (4°C) with the following primary antibodies: anti-METTL14 (Norvus), anti-CSNK1D (Norvus), anti-SLC35E1 (Norvus) and β-actin (Invitrogen). After washing, the membranes were further subjected to the appropriate secondary antibodies (Invitrogen). Blots were visualized by a ChemiDoc XRS system, followed by quantification using Image Lab software (Bio–Rad).

Free full text: Click here

Liu Q., Li Z., He L., Li K., Hu C., Chen J., Zhou F., Wang J., Li Y, & Xiao H. (2022). Molecular Characterization and Clinical Relevance of N6-Methyladenosine Regulators in Metastatic Prostate Cancer. Frontiers in Oncology, 12, 914692.

Publication 2022

PVDF membranes β-actin appropriate secondary antibodies ChemiDoc XRS system Image Lab software

Actin Antibodies Membranes Mettl14 Protein Pvdf Sds page

Corresponding Organization : Sun Yat-sen University Cancer Center

Other organizations : Chinese Academy of Medical Sciences & Peking Union Medical College

Top 5 similar protocols

Variable analysis

independent variables

Protein lysates

dependent variables

Protein expression levels of METTL14, CSNK1D, and SLC35E1

control variables

Protein concentration measurement using BCA Kit
SDS-PAGE resolution and PVDF membrane transfer
Overnight incubation of primary antibodies at 4°C
Washing of membranes
Incubation with secondary antibodies
Blot visualization using ChemiDoc XRS system
Quantification using Image Lab software (Bio-Rad)

positive controls

β-actin

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!