Blotting Techniques made easy
What you'll learn
- Part 1:Definition and introduction
- 3 types of Blotting techniques
- Sothern: 6 steps
- Part 2:Sothern Blotting explanation
- Part 3:Sothern Blotting 10 steps
- Part 4:Applications
- strengths and limitations
- Part 5: Northern Blotting
- steps: 8
- Part 6:Norther blotting applications
- advantages and disadvantages
- Part 7: Western blotting
- Data analysis
- Part 8:procedure
- Part 9:
- what is SNOW Drop
- No requirements
- Any Biology student can learn
Definition and introduction
3 types of Blotting techniques
Sothern: 6 steps
Sothern blotting Diagram
Sothern Blotting explanation
Sothern Blotting 10 steps
Membrane for bloat transfer
strengths and limitations
Norther blotting applications
advantages and disadvantages
Western blotting buffer preparation:
western blotting detection
what is SNOW Drop
Detecting Phosphorylation States of Proteins
Many proteins are post-translationally modified by kinases, which add phosphate groups to their substrates. Western blotting is great for detecting the presence of phosphorylated proteins. Phosphorylated proteins become heavier, due to the added weight of the phosphate group, so they often migrate more slowly than their un-phosphorylated forms.
Detecting Changes in Protein Levels Across Treatment Groups
Detecting the presence and absence of a protein may seem like a simplistic assay, but when each sample represents a different treatment group, western blotting can be very informative about the amount of each protein in treatment. This also applies to detecting how a certain treatment changes the post-translational modification of a protein (i.e. phosphorylation, ubiquitination, etc.).
Detecting Changes in Protein Levels Across Time Points
Western blotting is incredibly informative for determining the effect of time on a protein. For example, if each sample is a protein mixture of cells that are in different phases of the cell cycle, then western blotting will reveal how much a protein is present or absent during each phase.
Detecting Truncated Isoforms of Proteins
Many proteins are cleaved in order to be activated, or have naturally occurring truncation isoforms. Each isoform may have a different level of activity, a different target protein, or represent a different cellular state. Western blotting is great for detecting the ratio of truncated to normal isoforms of a protein.
Detecting Tagged Proteins
Some proteins are engineered, through the process of molecular cloning, to contain short sequences of amino acids that serve as a tag. Common tags include the HA-tag and the Myc-tag. These tags serve as a foreign protein epitope that does not naturally occur in the biological system being studied. Thus, the tag makes the protein easy to detect compared to all other naturally occurring proteins. An antibody directed to the tag will identify the presence and amount of the tagged protein in the western blot.
and many more in slide explanation
Who this course is for:
- Micro Biology
I have total 7+year of teaching experience in life sciences. I have educates students from different life science backgrounds like Botany, Zoology, Microbiology, Bio-chemistry and Biotechnology. I have extensive teaching experience in Bio-technology and related concepts. I worked as professors on Bio-technology, Bio-Chemistry and Botany.
can able to explain in practical manner and good demonstrative skills. any life science student can ask queries.