Western Blot Protein Quantification

Protein extraction and Western blotting were performed as previously described (31 (link)). Primary antibodies were diluted to 1:100 for CAV1 (Santa Cruz Biotechnology), 1:500 for COL1A2 (Novus Biologicals), 1:200 for THBS2 (Santa Cruz Biotechnology), and 1:5000 for beta actin (Sigma Aldrich). Anti-mouse (beta actin), anti-goat (THBS2), and anti-rabbit (CAV1, COL1A2) secondary antibodies were used at a concentration of 1:300 (Santa Cruz Biotechnology). Proteins were visualized by xerography on XOMAT™ AR film using an enhanced chemiluminescence system. Xerograms were digitized with an Epson flat-bed scanner and ImageJ software was used to quantify expression. A Student’s T-test was used to compare protein expression between treatment groups.

Partial Protocol Preview
This section provides a glimpse into the protocol.
The remaining content is hidden due to licensing restrictions, but the full text is available at the following link: Access Free Full Text.

Pekow J., Hutchison A.L., Meckel K., Harrington K., Deng Z., Talasila N., Rubin D.T., Hanauer S.B., Hurst R., Umanskiy K., Fichera A., Hart J., Dinner A.R, & Bissonnette M. (2017). miR-4728-3p functions as a tumor suppressor in ulcerative colitis-associated colorectal neoplasia through regulation of focal adhesion signaling. Inflammatory bowel diseases, 23(8), 1328-1337.

Publication 2017

CAV1 beta actin flat-bed scanner

Antibodies Beta actin Biologicals Cav1 Chemiluminescence Col1a2 Goat Mouse Novus Protein Rabbit Student Thbs2 Xerography

Corresponding Organization : University of Chicago

Other organizations : Northwestern University, University of Washington

Top 5 similar protocols

Variable analysis

independent variables

Treatment groups

dependent variables

Protein expression of CAV1, COL1A2, THBS2, and beta actin

control variables

Primary antibody dilutions (1:100 for CAV1, 1:500 for COL1A2, 1:200 for THBS2, and 1:5000 for beta actin)
Secondary antibody concentrations (1:300 for anti-mouse, anti-goat, and anti-rabbit)
Protein visualization method (xerography on XOMAT™ AR film using an enhanced chemiluminescence system)
Image analysis (digitized with an Epson flat-bed scanner and quantified using ImageJ software)

controls

Positive control: beta actin
Negative control: Not specified

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!