Lab Expectations

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.

Immunotherapy as an anti-neoplastic strategy has gradually claimed the spotlight over the past several years. In fact, some now consider it as one of the pillars of cancer therapy along with chemotherapy, radiation, surgery, and targeted therapy. Scientists and clinicians alike are cautiously optimistic, and point to response rates never seen before. 

And, why not take advantage of our own immune system and the best of its weaponry, the T cells, to fight cancer?

Journal Club: YAP-dependent reprogramming of Lgr5⁺ stem cells drives intestinal regeneration and cancer.

The article we have chosen was published in the October 29^th, 2015 issue of the journal Nature by Gregorieff et al., from the Lunenfeld Tanenbaum Research Institute and University of Toronto, Ontario Canada. The article is entitled YAP-dependent reprogramming of Lgr5⁺ stem cells drives intestinal regeneration and cancer, and focuses on stem cell dynamics within the intestinal epithelium, the most rapidly self-renewing tissue in adult mammals.

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.

The activity of genes and their regulatory elements is, in part, governed by their cell type-specific chromatin organization. Nucleosomes, the building blocks of chromatin, are wrapped around a core of histone proteins that are subject to many post-translational modifications that can either promote or silence gene expression.

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...

CRISPR, a nifty gene-editing tool, has swept the scientific community off its feet. It has a catchy name, it’s in every other publication, it is the subject of a volatile legal dispute involving claims to its intellectual property (1), its use has instigated a multitude of summits to do with its ethical ramifications and, despite the fact that it’s less than 4 years old, the Science journal named it the ‘breakthrough of the year’ (2). To give you some perspective, the other contenders were the New Horizon’s epic visit to the dwarf planet Pluto, the discovery of a lymphatic system in the brain, and the Ebola vaccine. 

Well, you get the picture, CRISPR is a big deal, but what on Earth is it and why are scientists so smitten by it?

Analysis of Immune Checkpoint Control Protein Co-expression in Breast and Ovarian Cancer Using Novel Rabbit Monoclonal Antibodies and Multiplex IHC

With an increasing number of biomarkers and, often, limited availability of biopsy material, there is a growing need for multiplexed assays for both research and clinical purposes. IHC based solutions are particularly attractive in the field of immuno-oncology, as maintaining spatial context within the tumor microenvironment provides meaningful and potentially actionable information.

Watch the video below.

CRISPR, a nifty gene-editing tool, has swept the scientific community off its feet. It has a catchy name, it’s in every other publication, it is the subject of a volatile legal dispute involving claims to its intellectual property (1), its use has instigated a multitude of summits to do with its ethical ramifications and, despite the fact that it’s less than 4 years old, the Science journal named it the ‘breakthrough of the year’ (2). To give you some perspective, the other contenders were the New Horizon’s epic visit to the dwarf planet Pluto, the discovery of a lymphatic system in the brain, and the Ebola vaccine. 

Well, you get the picture, CRISPR is a big deal, but what on Earth is it and why are scientists so smitten by it?

If you happened to attend AACR, AAI, or ASCO this year you’d agree that immune cell-based cancer therapies have caused quite the stir. Immunotherapy as an anti-neoplastic strategy has gradually claimed the spotlight over the past several years. In fact, some now consider it as one of the pillars of cancer therapy along with chemotherapy, radiation, surgery, and targeted therapy. Scientists and clinicians alike are cautiously optimistic, and point to response rates never seen before. 

And, why not take advantage of our own immune system and the best of its weaponry, the T cells, to fight cancer?