Posts tagged ‘Proteomics’

Largest Ever Map of Human Protein Interactions

One of the central questions in human biology is to understand how our genes determine which diseases we get and how severe they might be. Knowing just the DNA sequence, or the blueprint, is not enough. We must figure out how proteins, the genes’ products, work too.

Now an international team of researchers, jointly led by Dr. Fritz Roth (at Mount Sinai Hospital’s Lunenfeld-Tanenbaum Research Institute and the Donnelly Centre of the University of Toronto), and Dr. Marc Vidal (with the Dana-Farber Cancer Institute and Harvard Medical School in Boston), have produced the largest ever map of human protein interactions. This publicly available resource will be invaluable to anyone trying to understand complex genetic traits and develop new disease therapies.

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November 20, 2014 at 5:28 pm Leave a comment

Identifying hundreds of new connections in a critical growth pathway

Understanding the checks and balances that govern when and how much cells grow is key to understanding cancer. A study published in November 2013 by the Gingras lab uncovers pieces of the complex mosaic of molecular interactions or signals that govern the normal growth of cells, tissue, and organs.

Within all animal cells an important series of switches causes them to stop growing once a tissue has attained the right size. “This system is called the Hippo pathway because deregulation of this system leads to overgrowth, a ‘hippopotamus’ phenotype. The Hippo pathway consists of proteins that interact with one another, sense other control systems within our cells, and send signals to stop the cell growth,” says Dr. Gingras.

“Our study identified 749 interactions between proteins and enzymes that play a role in telling a cell when to stop growing. Of these, 600 have not been previously recognized in the Hippo pathway,” she says.

“These findings are promising because, to date, there are no drugs directed at the components of the Hippo pathway,” adds Dr. Jim Woodgett, Director of the Lunenfeld-Tanenbaum Research Institute. “Anne-Claude’s team’s work has added many new candidates for therapeutic intervention that may, for example, help in restricting the uncontrolled growth of tumour cells.”

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December 9, 2013 at 4:51 pm Leave a comment

Top 8 reasons for high background noise on your western blot

The following are the top 8 causes for overall high background on your western blot and potential solutions:

  1. Blocking was incomplete
    • Increase the concentration of blocker
    • Increase the duration of the blocking step
    • Use a different blocking agent
  2. Blocker was impure
    • Use a pure protein such as BSA or casein as a blocker
  3. Wash protocols were insufficient
    • Increase the number, duration, or stringency of the washes
    • Include progressively stronger detergents in the washes; for example, SDS is stronger than Nonidet P-40 (NP-40), which is stronger than Tween 20
    • Include Tween 20 in the antibody dilution buffers to reduce nonspecific binding
  4. The blot was left in the enzyme substrate too long (colorimetric detection)
    • Remove the blot from the substrate solution when the signal-to-noise level is acceptable, and immerse in diH2O
  5. Contamination occurred during electrophoresis or transfer
    • Discard and prepare fresh gels and transfer solutions
    • Replace or thoroughly clean contaminated foam pads if a tank blotter was used
  6. Excessive amounts of protein were loadedon the gel or too much SDS was used inthe transfer buffer. Proteins can pass through the membrane without binding and recirculate through a tank blotting system.
    • Reduce the amount of protein on the gel or SDS in the transfer buffer
    • Add a second sheet of membrane to bind excess protein
  7. The primary or secondary antibody was too concentrated
    • Increase antibody dilutions
    • Perform a dot-blot experiment to optimize working antibody concentration
  8. Incubation trays were contaminated
    • Clean the trays or use disposable trays

For more great information, download the protein blotting guide from Bio-Rad Laboratories.

Related Posts:
Which camera is best for chemiluminescent detection?
Four Great Tips for Effective Protein Blotting
Test your knowledge of protein blotting membranes
Efficient Electrophoresis and Protein Blotting: Aiding Advances in Cardiomyopathy Research
Protein blotting guide for novice and advanced users

November 2, 2012 at 1:10 pm Leave a comment

Do Proteins Have Their Own Social Network?

October 3, 2012 at 3:19 am Leave a comment

Revolutionizing Protein Research

Western blotting, the backbone of protein research, is a universal lab procedure. While the premise of a western blot is simple — target proteins are identified and quantitated via antibody-antigen interactions — the traditional workflow is labor intensive and time consuming. Researchers have long sought a faster solution — an archetype that would streamline the entire process of separation, transfer, and visualization of results without compromising data quality.

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June 27, 2012 at 10:10 am Leave a comment

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