1. Western Blot Lab

2. Western Blot reagents and equipment Mini Trans-Blot Apparatus : Passes electric current horizontally through gel – forcing negatively charged proteins to migrate out of gel onto nitrocellulose membrane Nitrocellulose membranes: Proteins bind to positive charge -Durable membrane can undergo multiple washing and incubation steps -White background to visualize color development of proteins

3. Western Blot reagents and equipment Blotting paper – supports gel and nitrocellulose -Protects filter pads during assembly and electrophoresis -Facilitates uniform flow of buffer and current through gel -100% cotton fiber – does not contain any additives that may interfere with blotting process

4. Western Blot reagents and equipment Filter pads: Press the gel and nitrocellulose tightly and uniformly -Eliminate bubbles – allows transfer of proteins out of gel onto the membrane -Should be cleaned in distilled water before use

5. Western Blot reagents and equipment Blotting buffer: Should be diluted to 1X Blocker: 5% nonfat dried milk powder in phosphate buffered saline (PBS) and 0.025% tween 20 Surface area unoccupied by proteins- blocked by incubating milk solution – before incubation with primary antibody

6. Western Blot reagents and equipment Primary antibody: recognizes or bind protein under study -Injecting chicken myosin protein into mice Secondary antibody: binds to primary antibody -Polyclonal goat anti-mouse antibody conjugated to horseradish peroxidase enzyme -Injecting goats with primary mouse antibodies

7. Western Blot reagents and equipment This project is funded by a grant awarded under the President’s Community Based Job Training Grant as implemented by the U.S. Department of Labor’s Employment and Training Administration (CB-15-162-06-60). NCC is an equal opportunity employer and does not discriminate on the following basis: against any individual in the United States, on the basis of race, color, religion, sex, national origin, age disability, political affiliation or belief; and against any beneficiary of programs financially assisted under Title I of the Workforce Investment Act of 1998 (WIA), on the basis of the beneficiary’s citizenship/status as a lawfully admitted immigrant authorized to work in the United States, or his or her participation in any WIA Title I-financially assisted program or activity.

8. Disclaimer This workforce solution was funded by a grant awarded under the President’s Community-Based Job Training Grants as implemented by the U.S. Department of Labor’s Employment and Training Administration. The solution was created by the grantee and does not necessarily reflect the official position of the U.S. Department of Labor. The Department of Labor makes no guarantees, warranties, or assurances of any kind, express or implied, with respect to such information, including any information on linked sites and including, but not limited to, accuracy of the information or its completeness, timeliness, usefulness, adequacy, continued availability, or ownership. This solution is copyrighted by the institution that created it. Internal use by an organization and/or personal use by an individual for non-commercial purposes is permissible. All other uses require the prior authorization of the copyright owner.