Antibody News

By Jamshed Arslan, Pharm. D., PhD.

Western blotting involves three steps: 1) separating proteins by size through gel electrophoresis 2) transferring separated proteins to a solid support (typically, a PVDF membrane) and 3) using antibody to mark your protein(s) of interest in the mixture of proteins. The second step of moving negatively charged proteins from the gel to a blotting membrane (PVDF) can be performed in dry or wet conditions.

Dry/semi-dry transfers

Dry conditions are famous for easiness and speed. Reagents, power packs and wiring involved in dry transfers are less messy and therefore, potentially less hazardous. They allow blotting of several...

By Jamshed Arslan Pharm.D., PhD.

Introduction

Kinome describes kinases, and kinomics refers to the kinase signaling. Studying the effects of reagent (exogenously applied growth factor or inhibitor) on kinase activity is a common experiment in cell, developmental and integrative biology laboratories. This blog offers a cost-effective and robust method of obtaining adequate amount of sample for kinomic analysis from mammalian culture cells like HEK cells after treatment with reagent. Using this method, you can easily get over 50 μg protein sample in 50 μl volume from a confluent well of a standard 6-well plate with 0.5 x 10⁶...

By Jamshed Arslan Pharm.D., PhD.

Myelin is a cholesterol-rich layer around nerves. Damage to this protective layer and the consequent slowing down of nerve impulses are hallmarks of multiple sclerosis (MS) and other demyelinating diseases. Myelin breakdown alters plasma levels of 27-hydroxycholesterol (27OHC) and other CNS-specific oxysterols, which are natural ligands of transcription factors called liver X receptors (LXRs). Since myelin phagocytosis is a source of intracellular oxysterols in the resident microglia and infiltrating macrophages, a team of scientists in Europe decided to test the hypothesis that myelin uptake and subsequent...

By Beth Melson, MS

Viral infection induces the production of reactive oxygen species (ROS) by inflammatory cells, the majority of which is produced by the NOX2 oxidase. While NOX2 oxidase contributes to the clearance of bacteria and fungi, its role in viral infection is less clear, and in fact seems to contribute to increased viral pathogenicity. Recently, it was demonstrated that viruses, including influenza A, induce NOX2 oxidase production through suppression of TLR7 sensing single-stranded RNA (ssRNA)¹. Therefore, inhibition of the NOX2 oxidase pathway represents an opportunity for viral...

By Michalina Hanzel, PhD

The multifaceted roles of the Wnt family of glycoproteins have been extensively characterized throughout embryonic development and adult homeostasis. The highly conserved, cell- and tissue- specific proteins orchestrate processes ranging from neural induction, cell proliferation and migration to adult neurogenesis and neuronal maintenance and regeneration. Recently, Wnt proteins have been proposed to have key roles in regulating pathological processes in the brain, especially those observed in neurodegenerative disorders, often resulting from pathological states of neuroinflammation.

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By Jody Bonnevier, PhD.

Flow cytometry: A Challenging Interdisciplinary Application

Flow cytometry has long been the cell analysis method of choice for immunologists, however more and more disciplines have recognized its powerful and flexible capabilities, and want to harness the advantages of this multiparameter single-cell analytical tool. Nowadays, the benefits of flow cytometry for cell phenotyping extend from basic to clinical research and diagnostic applications as well as to drug discovery and development.

In basic research, the same flow cytometry techniques used to phenotype T cell subsets can be applied to defining protein...

By Christina Towers, PhD.

Autophagy is highly conserved and tightly regulated process that all cell types use to recycle nutrients, particularly in the instance of stress¹. As a result, even small changes in signaling pathways, gene expression, or drug treatments can alter autophagy. If you stumble into the field of autophagy by happenstance and hypothesize a change in autophagy with your favorite cell manipulation of choice, you probably read a handful of publications and came to the conclusion that you should assay LC3 levels to test this hypothesis. In fact, LC3 expression can be a good readout for autophagy, but researchers should be...

By Jamshed Arslan Pharm.D.

Non-small cell lung cancer (NSCLC) is the most common type of lung cancer. NSCLC cells show increased expression of HIF-1alpha and HIF-2 alpha, which help cells overcome hypoxia. However, NSCLC samples have shown decreased mRNA levels of HIF-2 alpha-encoding gene called EPAS1. The mechanism of downregulation of EPAS1 transcription in NSCLC is unknown. A team of researchers from various institutes in Shanghai and Yangzhou, China, suspected transcriptional inactivation through DNA...

By Christina Towers, PhD.

Autophagy is a cellular recycling process and most often a pro-survival mechanism that regulates cellular homeostasis. On the contrary, apoptosis is an extensively conserved and elaborate programmed cell death process, and it is well established that the two processes are often opposing forces within the cell. Despite the agreed upon notion that autophagy can protect cells from apoptosis, the mechanistic link has yet to be elucidated. This is especially important given the 60 (and growing) number of clinical trials that are currently underway inhibiting autophagy in cancer in combination with other chemotherapies and targeted therapeutics.

The field is hopeful...

By Christina Towers, PhD.

In the last 20 years, epigenetic regulation has become front and center for almost all fields of biology and its role in diseases like cancer and neurodegeneration are being heavily studied. Epigenetics can be defined as a change in phenotype without a change in genotype that is caused by remodeling the chromatin landscape and is often mediated by changes in histone marks, like methylation, acetylation, phosphorylation, SUMOylation, ubiquitination, and glycosylation to name a few. The cellular recycling process of autophagy is among one of the many processes...

By Yoskaly Lazo-Fernandez, PhD

Introduction

Evidence has accumulated recently indicating that common cancer therapies might stimulate metastasis in a significant number of cancer patients¹. In fact, neoadjuvant chemotherapy (NAC) drugs, which are administered preoperatively as a first line of treatment, have been associated with increased infiltration of endothelial progenitor cells and macrophages in primary tumors, resulting in higher angiogenesis and tumor regrowth². Mechanisms by which these cellular injury responses induce metastasis, both in preclinical in vivo models, and in human...

By Christina Towers, PhD.

Parkinson's disease is a debilitating neurodegenerative condition defined by the accumulation of alpha-synuclein-containing (alpha-SYN) intra-cytoplasmic inclusions, called Lewy bodies. The protein degradative processes of autophagy, and most specifically chaperone mediated autophagy and mitophagy, play an important protective role against this disease by targeting neurotoxins for lysosomal mediated degradation¹. Alpha-SYN neurotoxins most commonly affect neurons, but they can also...