This video is about western blot analysis with ImageJ, quantifying bands on SDS-PAGE using ImageJ, ImageJ tutorial, Using ImageJ to quantify proteins bands, western blot densitometry in ImageJ, quantifying intensity in imagej, imagej quantify gel bands, western blotting, western blot data, blot data, western data, or Immunoblotting. Hopefully it helps if you're new to the topic/technique. This channel takes you through some of the techniques and concepts I've learnt working as a Research Assistant.
I’ve worked in medical research for years and want to be useful to people new to the lab life. I'm Adwoa (Adwoa Biotech), a Biotechnology graduate. PREVIOUS VIDEO USING INTEGRATED INTENSITY IN IMAGEJ: As each peak is selected, a value is given for that peak.Ĭopy the values and analyse in excel or another spreadsheet program as shown in the previous video. use the ‘wand’ tool to select each of the peak. use the ‘line’ tool to mark the base of each peakĨ. Divide IntDen for protein of interest by IntDen for beta-actin or house-keeping gene. 10.Save the table (will open in Excel) 11.Normalize the intensity: a. Hit M on the key board after selecting the area to see the measurements. This shows you individual peaks associated with each protein band in the lane.ħ. Select area you want to measure (keep it tight) 9. Move the rectangle to all additional lanes and SELECT NEXT LANE after each move to register the lane go back to ANALYZE - GELS - SELECT NEXT LANE Use the forward arrow of your keyboard to move to the next lane.ĥ. Go to ANALYZE - GELS - SELECT FIRST LANEĤ. draw around the lane you wish to quantifyģ. The method is also preferred when the bands you need to quantify are very faint.Ģ. That is, different proteins that cannot be quantified together as an Integrated Density. This method is useful when you have multiple western blot bands that are distinct from each other. In this video I show the NIH-recommend process for quantifying bands derived from SDS-PAGE.īelow are the instructions for using the area under the peak method in your western blot quantitation. ImageJ is a public domain program developed by Wayne Rasband while at the National Institutes of Health (NIH). Department of the Treasury, Office of Foreign Assets Control.Subscribe for a fun approach to learning lab techniques: export laws and regulations, contact the U.S. Department of Commerce).Īll rights to use the Bio-Rad product(s) are granted on condition that such rights are forfeited if You fail to comply with the terms of these Export Compliance Requirements.įor more information regarding U.S. (For a current list of all sanctioned and/or embargoed persons or entities, please consult the U.S. Department of Commerce as a sanctioned and/or embargoed person or entity. transfer, export or re-export, directly or indirectly, any Bio-Rad product, technology or software (or any part thereof, or the product of such technology or software, or any process or service which is the product of such technology or software) to any person or entity identified by the U.S.transfer, export or re-export any Bio-Rad product to Your customers or any intermediate entity in the chain of supply if any Bio-Rad product will be used in the design, development, production, stockpiling or use of missiles, chemical or biological weapons or for nuclear end uses or.sell, license, or otherwise provide or ship, directly or indirectly, any Bio-Rad product or technical data (or the direct product thereof) for export or re-export to the embargoed or restricted* countries (Cuba, Iran, *North Korea, Sudan, and *Syria) or.use any Bio-Rad product if You are located in, under the control of, or a national or resident of any embargoed or restricted* countries (Cuba, Iran, *North Korea, Sudan, and *Syria) or.You agree that, unless You receive prior authorization from the United States government, You and/or Your organization shall not: Export-Re-export Requirements and Enhanced Proliferation Control Initiative. export control laws, including but not limited to the U.S. Export of any Bio-Rad product or technical information relating to the product (i.e., manuals, memoranda, specifications, schematics, etc.) outside of the United States is subject to U.S.