Lab Expectations

Type 2 diabetes is a growing epidemic, and is recognized as one of the most serious metabolic disease worldwide. A multifactorial disease, type 2 diabetes is a perfect example of metabolic miscommunication between different organs resulting in a pathological outcome. According to CDC in the United States, 29.1 million people in the United States have diabetes, and 8.1 million may be undiagnosed. The disease affects more than 1 in every 10 adults, and seniors aged 65 and above are most affected. What makes the disease morbid are the secondary complications associated with it; atherosclerosis and cardiomyopathy are the leading cause of death in people diagnosed with type 2 diabetes. The need for an effective treatment has become a global health priority.

Reuben Shaw, Ph.D

Professor, Molecular and Cell Biology Laboratory, Deputy Director, Salk Cancer Center, The Salk Institute for Biological Studies

AMPK, a highly conserved sensor of cellular energy status, is found in all eukaryotic cells and maintains metabolic homeostasis by reprogramming growth, metabolism, and autophagy in the face of cellular stresses.

Traditionally, flow cytometry has been used to identify distinct cell types within a heterogeneous pool of cells, based on extracellular or surface marker expression, an application commonly known as immuno-phenotyping. However, this technology is also readily amenable to intracellular target detection and can be successfully applied to the study of complex signaling events.

Specificity, consistency, and optimized assay conditions are three key elements that help ensure reliable immunofluorescence (IF) staining results each and every time. So, before starting an experiment you should ask yourself the following three questions:

• Is your antibody specific?
• Is your antibody supported by an optimized IF protocol?
• Is your antibody performing consistently?

Let's take them each in turn...

Specificity, consistency, and optimized assay conditions are three key elements that help ensure reliable immunofluorescence (IF) staining results each and every time. So, before starting an experiment you should ask yourself the following three questions:

• Is your antibody specific?
• Is your antibody supported by an optimized IF protocol?
• Is your antibody performing consistently?

Let's take them each in turn...

Reuben Shaw, Ph.D

Professor, Molecular and Cell Biology Laboratory, Deputy Director, Salk Cancer Center, The Salk Institute for Biological Studies

AMPK, a highly conserved sensor of cellular energy status, is found in all eukaryotic cells and maintains metabolic homeostasis by reprogramming growth, metabolism, and autophagy in the face of cellular stresses. AMPK is activated by direct binding of AMP and ADP to its regulatory subunits, which enhances its phosphorylation by the upstream kinase LKB1, a tumor suppressor gene frequently inactivated in sporadic human lung and cervical cancer.

Traditionally, flow cytometry has been used to identify distinct cell types within a heterogeneous pool of cells, based on extracellular or surface marker expression, an application commonly known as immuno-phenotyping. However, this technology is also readily amenable to intracellular target detection and can be successfully applied to the study of complex signaling events.