Antibody News

RNA-binding protein Staufen1 conspires with Atxn2 in stress granules to cause neurodegeneration by dysregulating RNA metabolism

Tuesday, December 11, 2018 - 11:19
PCP4 expression in mouse cerebellum Purkinje neurons, IHC

By Jamshed Arslan Pharm.D.

Spinocerebellar ataxia type 2 (SCA2) is a movement disorder characterized by neurodegeneration. The cause of this autosomal dominant disease is a mutation in the RNA processing gene Atxn2, which leads to polyglutamine (polyQ) expansion in ATXN2 protein. The association between mutant polyQ proteins, neurodegeneration, and dysfunctional autophagy has been extensively studied in amyotrophic lateral sclerosis (ALS), autism, and Huntington’s disease, but not in SCA2. It is noteworthy that ATXN2 is widely expressed in our nervous system, partly as a constituent of stress granules (SGs), which...

Flow Cytometry Basics for the Non-Expert

Tuesday, December 4, 2018 - 10:35
Flow cytometry dot plots, histograms, density plots

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...

Losing memory: Toxicity from mutant APP and amyloid beta explain the hippocampal neuronal damage in Alzheimer's disease

Tuesday, November 27, 2018 - 13:42
Map2 and Fox3/NeuN expression in rat hippocampus, IHC

By Jamshed Arslan Pharm.D.

Alzheimer's disease (AD) is an irreversible brain disorder that destroys memory and thinking skills. The telltale signs of AD brains are extracellular deposits of amyloid beta (a polypeptide that comes from proteolysis of precursor protein APP) and intracellular neurofibrillary tangles. The consequent neurodegeneration affects regions associated with memory and cognition, the most noteworthy of which is the hippocampus. It goes without saying that generating immortalized primary hippocampal neurons that could express AD-related protein(s) would immensely...

Application Focus: I see an increase in LC3, now what?

Monday, November 19, 2018 - 13:05
LC3A expression in HeLa cells, chloroquine treated, ICC

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...

E-syt in Autophagosome biogenesis: What is the source of it all?

Tuesday, November 13, 2018 - 13:44
Calreticulin expression in vesicles and endoplasmic reticulum of HeLa cells, ICC

By Christina Towers, PhD.

Macroautophagy is a cellular recycling process that requires the formation of double membrane structures to engulf and degrade damaged cytoplasmic material. The pathway involves over 20 core autophagy proteins (

Negative feedback regulation of EPAS1 gene in non-small cell lung cancer through DNA methyltransferases

Tuesday, November 6, 2018 - 09:29
HIF-1 alpha expression in human lung tissue, IHC

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...

Nickel induces migratory and invasive phenotype in human epithelial cells by epigenetically activating ZEB1

Tuesday, October 30, 2018 - 09:05
Epigenetic mechanisms Novus Biologicals

By Jamshed Arslan Pharm.D.

Nickel (Ni) is a naturally abundant metallic element. It is a major component of stainless steel, coins, and many other items of daily use. Disturbingly, Ni exposure is associated with cancer and several diseases of the lung, kidney and cardiovascular system. Despite being carcinogenic, Ni’s ability to cause DNA mutations and induce oxidative stress is low. To explain this paradox and investigate how the impact of carcinogen exposure may last for months or even years, researchers at the New York University and University of Virginia examined Ni-induced changes in transcription and cellular regulation in human epithelial cells from lung and urinary...

A link between Autophagy and Apoptosis: Chat with First Author Brent E. Fitzwalter

Tuesday, October 23, 2018 - 09:00
Apoptosis poster Novus Biologicals

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...

The inflammasome: an inflammation-initiating machine, Novus Biologicals

Wednesday, October 10, 2018 - 09:34
ASC/TMS1 expression in Hela cells ICC

By Stephanie Melchor

The inflammasome is a large, multimeric protein complex found primarily in innate immune cells, which are white blood cells that can attack a wide range of pathogenic threats. Three main elements make up the inflammasome: 1) the sensor – a cytosolic pattern recognition receptor that senses pathogen- or damage-associated molecular patterns (PAMPs and DAMPs), 2) an adaptor protein called ASC, and 3) a caspase effector that cleaves and activates pro-IL-1 beta and pro-IL-18.

Prior to inflammasome activation, a cell must first be "primed" with...

Epigenetic Control of Autophagy

Tuesday, October 9, 2018 - 08:48
G9a expression in human MCF7 cell line ICC

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...

Bad news for stomach cancer: BAMBI protein inhibits gastric carcinoma via TGF-beta/epithelial-mesenchymal transition signaling

Tuesday, October 2, 2018 - 09:03
Collagen-I expression in rat colon tissue IHC

By Jamshed Arslan Pharm.D.

Gastric carcinoma is the second leading cause of cancer-related deaths worldwide. One of the key features of gastric carcinoma is acidosis, which promotes growth and metastasis of gastric cancer cells by inducing HIF-1alpha, TGF-beta, and epithelial-mesenchymal transition (EMT) signaling pathways. Gastric cancer cells have a reduced expression of bone morphogenetic protein and activin membrane-bound inhibitor (BAMBI), a pseudoreceptor that negatively regulates TGF-beta signaling by inhibiting the...

Chemotherapy-induced metastasis: An unexpected foe?

Tuesday, September 25, 2018 - 09:02
Mena plasma membrane, cytosol and focal adhesion expression in human cell line U-2 OS ICC

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...

Advancing CAR T Therapies with CRISPR/Cas9

Tuesday, September 18, 2018 - 09:27
Expression of CD4 and CD8 in peripheral blood mononuclear cells ICC

By Victoria Osinski

Scientists have turned to gene editing techniques to modify patients’ T cells to combat cancer, but are often limited by factors including cost, low cell yields, or availability of expertise for therapy development. Interest lies in developing “off-the-shelf”, universal, donor cells, which require thorough T cell modifications to make this approach safe and feasible. Currently, scientists are leveraging the use of the gene editing tool clustered regularly interspaced short palindromic repeats (CRISPR)/Cas9 system to advance CAR T therapies.

"Off-the-shelf" donor T cells requires a combinatorial approach

Patients...

How a cell "reaches" out for help

Tuesday, September 11, 2018 - 09:30
Expression of alpha-synuclein in rat neuron-glial cultures ICCchloroquine-treated, ICC

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...

Combined NOTCH/MAPK inhibition: A promising new anticancer approach

Tuesday, September 4, 2018 - 11:23
MAPK signaling pathway Novus Biologicals

By Yoskaly Lazo-Fernandez, PhD

Introduction

Cancer invasion requires cancerous cells to overpower the barriers of the extracellular matrix and reach adjacent tissues. Invading cells undergo a process called epithelial-mesenchymal transition (EMT), which enables them to dedifferentiate into a highly invasive mesenchymal stem cell-like phenotype. Interestingly, once the cells have reached their new location through metastasis, they undergo a reverse process named mesenchymal-epithelial transition (MET) which allows the solid tumor to reorganize in the distant organ.

Molecular pathways involved in cancer invasion have thus become important targets for anticancer...

Rescuing motor neuronal loss in amyotrophic lateral sclerosis by inhibiting phosphatidylinositol-4 kinase

Tuesday, August 28, 2018 - 08:58
Novus C. elegans antibodies

By Jamshed Arslan Pharm.D.

Motor neurons control our ability to speak, walk, swallow, and even, breath. People with a motor neuron disease called amyotrophic lateral sclerosis (ALS) cannot perform these primal functions optimally. A point mutation (P56S) in VAMP associated protein isoform B (VAPB), a protein associated with the membrane around the endoplasmic reticulum, causes late-onset familial ALS8 by prompting VAPB to aggregate and lose its job of preventing buildup of abnormal proteins. To study ALS8, C. elegans is one of the animals of choice due to its simple neurophysiology: a single worm has only 302 neurons and 113 of them are...

Measuring Autophagic Flux with LC3 protein levels: The do's and don'ts

Tuesday, August 21, 2018 - 08:50
Expression of p62/SQSTM1 in HeLa cells mock- or chloroquine-treated, ICC

By Christina Towers, PhD.

Autophagy is a dynamic cellular recycling process that can be influenced by many different external and internal stimuli. The most commonly used assay to measure autophagy is a western blot for the autophagic membrane-specific protein, LC3. While this assay is relatively simple, if not controlled properly, the results can easily be misinterpreted. Indeed, the most accurate way to measure autophagy is with an autophagic flux assay defined as the new formation of double-membrane vesicle-like structures called autophagosomes and their...

Knockout Validation and the Reproducibility Crisis

Tuesday, August 14, 2018 - 09:34
Vimentin expression in K562 leukemia cell line ICC

By Colin O'Neill, 2018 Novus Intern

The specificity and affinity of an antibody for its target protein—and only its target is valued as an indicator of its experimental reliability. The frustrating ability of an antibody to bind to multiple proteins has prompted a reproducibility crisis in the life science community. Specifically, if an antibody varies in specificity between batches, attempts to reproduce significant findings with that antibody may fail. With samples, time, and funding at stake, scientists have developed novel validation techniques to ensure antibodies solely bind to their intended targets to avoid false positives and ambiguous results. Nature's 2015 article...

Eat responsibly: Epigenetic downregulation of Ankrd26 gene by long-term high-fat intake promotes obesity and inflammation

Tuesday, August 7, 2018 - 08:38
DNTM3A expression in human cell line A-431 ICC

By Jamshed Arslan Pharm.D.

White adipose tissue (WAT) represents the primary site to store energy in humans. WAT’s endocrine regulation of energy balance is controlled by nutritional status, exercise, and hormones like insulin. Partial inactivation of a gene highly expressed in WAT, called Ankrd26, induces obesity and diabetes in mice. Moreover, feeding mice high-fat diet (HFD) has been suggested to cause Ankrd26 promoter hyper-methylation, an epigenetic mechanism of gene silencing. Working along these lines, a team of researchers from Italy and USA set out to elaborate how HFD can epigenetically...

You complete me: Natural killer cells need TGF-beta inhibition to effectively combat cancers

Tuesday, July 31, 2018 - 08:22
TGF-beta signaling pathway

By Jamshed Arslan Pharm.D.

Natural killer (NK) cells are lymphocytes of the innate immune system that were first discovered for their “natural” ability to kill cancer cells. To use NK cells as anti-cancer therapy, they are co-cultured with feeder cells that are sensitive to attack by NK cells. The killing of feeder cells in turn generates highly efficient tumor-killing NK cells. However, a major challenge in using these ex vivo expanded NK cells as anti-cancer therapy is the highly immunosuppressive tumor microenvironment, partly because the cancer cells, myeloid derived suppressor cells and other stromal cells produce excessive amounts of...

Meningeal lymphatics: recent discovery defying the concept of central nervous system 'immune privilege'

Tuesday, July 24, 2018 - 08:57
Regulation Of Lymphangiogenesis Pathway Bioinformatics

By Jennifer Sokolowski, MD, PhD.


Identification and characterization of meningeal lymphatics

The recent discovery of a lymphatic system in the meninges surrounding the brain and spinal cord has spurred a surge of interest in and has redefined our understanding of immunity in the central nervous system (CNS).1,2 The lymphatic system in the brain is composed of the glymphatic system and meningeal lymphatic vessels.3 The glymphatic system involves convective flow of cerebral spinal fluid and interstitial fluid and uses para-arterial flux to clear solutes and metabolites from the brain parenchyma. In concert, the meninges harbor bona...

Monitoring Autophagy in Neurons

Tuesday, July 17, 2018 - 08:49
Beclin 1 expression in human cerebellum, Purkinje cells IHC

By Christina Towers, PhD.

Autophagy is a critical cellular process used by most cells in the body to recycle nutrients and prevent harmful buildup of damaged proteins. It is particularly important in the brain, where a handful of pathologies have been linked to autophagy dysregulation. Conditional neuronal knock out of the core autophagy gene, ATG7, results in viable mice that eventually succumb to neurodegeneration accompanied by an accumulation of ubiquitin protein aggregates1, 2. Likewise, decreased levels of functional autophagy have been linked to the three most common neurodegenerative diseases including...

Antigen-loss relapse after successful CAR-T therapy; What do we do now?

Tuesday, July 10, 2018 - 08:30
Acute lymphoblastic leukemia disease bioinformatics

By Jacqueline Carrico, BS, MD

Tumor cell mechanisms driving tolerance to CAR-T

Despite very promising results of CAR-T therapy in acute lymphoblastic leukemia, B-ALL, antigen-loss relapse has arisen as a major challenge for maintaining long term remission. Here we will review the potential mechanisms for antigen-loss relapse and some novel strategies to combat this problem.

One possible mechanism for antigen loss is downregulation of expression of the target protein in the malignant cells. Constant therapeutic stress from the highly-targeted CAR T-cells may lead to this phenomenon. Additionally, tumor...

Developmental regulator Daam2 promotes glial cell tumors by degrading Von Hippel-Lindau protein

Tuesday, July 3, 2018 - 08:35
GFAP expression neuron glia mixed culture ICC

By Jamshed Arslan Pharm.D.

Glioblastoma is an aggressive type of cancer that forms from the star-shaped glial cells of the central nervous system, called astrocytes. Intriguingly, several genes linked to glioblastomas, including the gene for the tumor suppressor Von Hippel-Lindau protein (VHL), do not exhibit mutations. VHL prevents tumorigenesis by binding and modulating the function of hypoxia-inducible factor HIF-1 alpha and hydroxylated serine-threonine kinase Akt. Developmental processes that maintain cells in a...

MAPK Signaling Links Autophagy and Inflammation

Tuesday, June 26, 2018 - 09:53
Autophagy signaling pathway

By Christina Towers, PhD.

MAPK Signaling and Disease

Mitogen-activated protein kinases (MAPKs) are serine-threonine kinase signaling molecules that function in a variety of cellular processes including cell differentiation, proliferation, survival, death, and transformation.  In mammals, there are 3 subfamilies of MAPKs including ERK, p38, and c-Jun each containing multiple isoforms. The MAPK signaling cascade is mediated by an upstream MAP3K which phosphorylates and activates a MAP2K which in turn activates a MAPK...

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