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Antibody News

Deriving neural precursor cells from human induced pluripotent stem cells

Tuesday, July 16, 2019 - 14:16
Sox2 antibody expression in induced pluripotent stem cells, ICC

By Jennifer Sokolowski, MD, PhD.

Human induced pluripotent stem cells (iPSCs) can be used to create models of human organ systems and are useful for a) ascertaining the mechanisms underlying pathological conditions and b) developing and testing therapeutics. For example, studies have used iPSCs from human patients with diseases like Alzheimer's and Parkinson's to test the influence of pathological isoforms of proteins as well as the efficacy of genetic rescue.1,2

Optimized protocols to turn induced pluripotent stem cells...

Stemness is responsible for onset and metastasis of colorectal cancer

Tuesday, July 9, 2019 - 09:58
beta-Catenin expression in mouse colon, IHC

By Jamshed Arslan, Pharm. D., PhD.

Colorectal cancer stem cells are a rare subpopulation of colorectal cancer cells that can self-renew and initiate and sustain tumor growth when transplanted into an animal host.1,2 Colorectal cancer stem cells are identified by CD133, CD166, CD44, nuclear-beta-catenin, ALDH1, EphB2 and...

PAMPs and DAMPs: What is the Same and What is Different About These Molecules?

Wednesday, June 26, 2019 - 14:33
TLR interactive signaling pathway, Novus Biologicals

By Victoria Osinski

What are PAMPs and DAMPs

Inflammation results from stimuli signaling damage or infection. The immune system inflammatory response can be beneficial or harmful depending on the type and duration of stimuli. The source, structure, and abundance of these stimuli vary quite a bit. One major category of inflammatory stimulation, or "signal 0s" is the family of pathogen-associated molecular patterns (PAMPs) and damage-associated molecular patterns (DAMPs).1,2 These patterns are found on bacterial cell walls, DNA, lipoproteins, carbohydrates, or other structures. While many DAMPs and PAMPs have been identified, they stimulate...

Autophagic flux: Is p62 a good indicator?

Tuesday, June 25, 2019 - 09:26
p62 detection via IHC-Paraffin, Novus Biologicals

By Christina Towers, PhD

Is p62 a good indicator of autophagic flux? The short answer: Yes … but … SQSTM1 encodes the cargo adaptor protein, p62, which interacts with autophagic substrates and delivers them to autophagosomes for degradation. In the process, p62 is itself degraded and when autophagy is induced, a corresponding decrease in p62 levels is observed. Congruently, when autophagy is potently inhibited, i.e. genetic knock out of ATG core autophagy proteins like ATG5 or ATG7, p62 accumulates in the cell. These changes can be observed by immunofluorescent staining, western blotting, and immunohistochemistry...

Identifying tumoral and stromal transcriptomes that underlie tumor plasticity and stromal neuroinflammatory response in brain metastasis

Tuesday, June 18, 2019 - 09:45
AIF-1/Iba-1 expression in mouse brain, IHC

By Jamshed Arslan, Pharm. D., PhD.

Cancers in the brain often come from tumors elsewhere in the body. Several adaptive mechanisms influence brain metastasis, such as blood brain barrier leakage that can be induced by stroma or disseminated tumor cells. The brain tumor microenvironment is complex, but the metastasizing cancer cells share common molecular features. To define such molecular characteristics of brain metastasis in vivo, a team led by Dr. Don Nguyen of Yale University School of Medicine developed an advanced RNA sequence-based approach....

Mechanisms of Neurodegeneration: Neuroinflammation and microglial activation

Tuesday, June 11, 2019 - 10:51
Alzheimer's Disease and Neuroinflammation Poster, R&D Systems, showing blood vessle and both health and diseased neurons.

By Michalina Hanzel, PhD

In this final instalment of our three blog-posts series on major cellular mechanisms responsible for neurodegenerative disorders, we will explore the processes of neuroinflammation and microglial activation. Previously, the role of autophagy in the clearance of aggregation-prone proteins in the context of neurodegenerative diseases was discussed in...

Best genes to knockout to control autophagic flux

Tuesday, June 4, 2019 - 08:27
Autophagy interactive signaling pathway, Novus Biologicals, macroautophagy induction, autophagosome maturation, microautophagy, autolysosome, elongation

By Christina Towers, PhD

Autophagy flux: Basic principles

Autophagic flux is defined as the amount of cellular material degraded and recycled through the process of autophagy, whereby cells break down and discard waste. Briefly, the autophagy process involves the formation of an autophagosome, which engulfs cytoplasmic materials. Fusion of the autophagosome with acidic lysosomes mediates the degradation of cellular contents. The process involves over 30 core autophagy proteins (...

Repurposing FDA-approved drugs to combat the rise of antibiotic resistance

Tuesday, May 28, 2019 - 09:03
Actin staining with Phalloidin with VECTASHIELD(R) Hard Set(TM) Antifade Mounting Medium, Novus Biologicals

By Beth Melson, MS

Antibiotic resistance is a global threat to public health. Widespread, inappropriate use of antibiotics, such as to treat viral infections or promote growth in livestock, has led to increased incidence of antibiotic resistance1. Resistance is costly to both human health as well as the economy1, and thus the identification of new, effective antibiotics is a public health priority. One solution to this problem is to repurpose commercially available, FDA-approved drugs to target host cell processes that would...

Mechanisms of Neurodegeneration: Mitochondrial Dysfunction and Oxidative Stress

Tuesday, May 21, 2019 - 07:44
3D image of dopaminergic neurons shown in green, Bio-Techne.

By Michalina Hanzel, PhD

In this second installment of our three blog-posts series on major cellular mechanisms responsible for neurodegenerative disorders, we will explore the processes of mitochondrial dysfunction and oxidative stress. Previously, the role of autophagy in the clearance of aggregation-prone proteins in the context of neurodegenerative diseases was discussed in Mechanisms of Neurodegeneration:...

Toll-like receptor 2 activation contributes to oral squamous cell carcinoma development and miRNA-mediated drug resistance

Tuesday, May 14, 2019 - 08:56
Nuclear Factor kappa B (NFkB) signaling pathway poster, Novus Biologicals

By Jamshed Arslan, Pharm. D., PhD.

Squamous cell carcinoma is the most common cancer in the oral cavity.1 The tumor surface biofilms in oral cancers contain high levels of aerobic and anaerobic microorganisms.1,2 Periodontal pathogens can activate epithelial Toll-like receptors (TLRs) on tumor cells to promote oral squamous cell carcinoma (OSCC) progression. In this regard, TLR2 is known to recognize micro-organisms in the oral cavity by forming a heterodimer with TLR1 or TLR6...

Make each cell count: How to assess autophagy using flow cytometry

Tuesday, May 7, 2019 - 09:32
LC3B expression in HeLa cells treated with chloroquine overnight, ICC. Novus Biologicals

Kristy R. Howell, PhD

The cellular recycling process known as autophagy may be induced by a variety of conditions including reduced nutrient availability, serum starvation and pharmacological agents (e.g., Rapamycin (mTOR)). Upon autophagy induction, a process of increased sequestration of dysfunctional cellular components into autophagosomes ensues. The autophagosome fuses...

Mechanisms of Neurodegeneration: Protein aggregation and failure of autophagy

Tuesday, April 30, 2019 - 09:54
Alzheimer’s disease video Novus Biologicals

By Michalina Hanzel, PhD

In a series of three blog posts I will briefly explore the major cellular mechanisms responsible for many neurodegenerative disorders. The first, and perhaps the most apparent, is the accumulation of misfolded, aggregate-prone proteins, as a result of a failure of the autophagy system, which leads to the loss of neuronal structure and function and eventual neuronal loss.

Autophagy is a conserved cellular process used to eliminate obsolete proteins and cell components by degradation and...

Success of combined IL-10 and IL-12 therapy in colon cancer depends on IFN-gamma and gut barrier integrity

Tuesday, April 23, 2019 - 08:28
IL10 receptor alpha expression in human colon carcinoma, IHC

By Jamshed Arslan, Pharm. D., PhD.

Colon cancer is responsible for over 600,000 deaths per year worldwide. Colon cancer can be classified into two categories: mismatch repair (MMR)-deficient and MMR-proficient cancers.1 MMR-deficient colon cancer shows high CD8+ cytotoxic T-lymphocyte (CTL) and low Th17 cells, while MMR-proficient colon cancer shows an opposite trend. MMR-proficient colon cancer is associated with poor prognosis and is resistant to anti-cancerous immune checkpoint inhibitors. In other words, outcome of immune therapy is...

How to visualize autophagy by microscopy

Tuesday, April 16, 2019 - 09:36
LC3B  and alpha tubulin expression in HeLa cells treated with Chloroquine, ICC, immunocytochemistry, immunofluorescence

By Christina Towers, PhD

Autophagy is a recycling process that relies on the formation of a unique organelle termed an autophagosome. An elegant way to monitor autophagy is through various microscopy techniques to visualize and quantify the formation and turnover of these double membrane structures. Some techniques are more quantitative than others, each accompanied by advantages as well as caveats.


Identifying the autophagosome with Transmission Electron Microscopy (TEM)


This technique is the only tool that can reveal all of the dynamic stages of a...

How To Identify B Cell Subsets Using Flow Cytometry

Tuesday, April 9, 2019 - 08:49
Explore new Flow Cytometry Handbook, Bio-Techne

By Victoria Osinski

Using Flow Cytometry to Identify B Cell Subsets

Identifying cellular subsets by flow cytometry requires careful and thorough planning in order to ensure the correct subset of cells are identified and isolated. While generally thought of as antibody-secreting cells, an expanding body of research proves that B cells are a heterogeneous population. Flow cytometry can be used to identify many of these B cell subsets, so long as important nuances and technical approaches are considered.

Setting Up to Identify B Cell Subsets

Before subsetting B cells, steps must be...

Lipopolysaccharide from gut microbiome localizes in human atherosclerotic plaques and promotes TLR4-mediated oxidative stress

Tuesday, March 26, 2019 - 09:05
TLR signaling pathway, Novus Biologicals

By Jamshed Arslan, Pharm. D., PhD.

Atherosclerosis is a chronic inflammatory condition in which plaques of fats and other substances slowly buildup on the inner walls of arteries to restrict blood flow. In atherosclerosis-associated ailments like obesity, hypertension and type II diabetes, intercellular tight junctions are impaired, leading to increased gut permeability. This means that products of intestinal microbiota like lipopolysaccharide from Gram-negative bacteria can seep into the blood stream.

A pro-atherogenic role of lipopolysaccharide has been suggested. For example,...

Toll-like receptors in the intestinal epithelial cells

Tuesday, March 19, 2019 - 10:30
Defensin alpha 5 expression in human small intestine, IHC

By Jamshed Arslan, Pharm. D., PhD.

Toll-like receptors (TLRs) are microbe-sensing proteins that act as first responders to danger signals. TLRs help the intestinal epithelial cells (IECs) recognize commensal bacteria from the harmful/foreign ones. Polymorphisms and variants of TLRs have been linked with immune system’s attack on commensal bacteria, leading to inflammatory bowel disease and colorectal cancers. The endogenous pattern of TLR expression in IECs and the functional outcome of TLR signaling in vivo are not well defined due to various technical difficulties. Thanks to the researchers in institutes in California and...

Probiotics and exercise can slow Alzheimer’s progression by altering composition of gut bacteria

Tuesday, March 12, 2019 - 09:11
beta amyloid expression in human Alzheimer’s disease hippocampus, IHC

By Jamshed Arslan, Pharm. D., PhD.

Gut microbiome modulates various disorders even outside the gut. For example, the microbiome is linked to amyloid-β deposition and the consequent inflammation in Alzheimer’s dementia. Altering gut microbiome through probiotics and/or exercise has been hypothesized to improve memory in Alzheimer’s disease (AD). A multi-continental collaboration of scientists tested this hypothesis on AD mice and found that probiotics and exercise can retard AD progression, at least partly by changing the microbiome.

Probiotics with exercise improve memory in AD mice

The researchers used Framelim, a product...

Microglia: pruning shears for homeostatic maintenance in the brain

Tuesday, March 5, 2019 - 08:58
Iba1 expression in mouse fore brain, IHC

By Jennifer Sokolowski, MD, PhD.

Microglia play a critical role in pruning neurons and synapses during homeostatic maintenance in the adult brain.1 A recent study by Ayata et al. (2018) identified regional differences in clearance and found a role for epigenetic control via polycomb repressive complex 2 (PRC2) in adult rodents.2

How to evaluate microglial clearance?

Ayata et al. defined a cell clearance phenotype by labeling microglia with iba1 and CD68 and evaluating morphology, cell volume, and CD68+ lysosome content. They also...

The LC3 A, B, C’s and 1, 2, 3’s

Tuesday, February 26, 2019 - 13:38
Atg5 expression in HeLa cells, ICC, immunocytochemistry, immunofluorescence

By Christina Towers, PhD

Autophagy is a catabolic process used to breakdown and recycle damaged proteins and organelles. It is a multistep process that, in its simplest form, consists of 4 steps: initiation, phagophore nucleation, elongation, and fusion – resulting in a double membrane structure called an autophagosome that, upon fusion with the acidic lysosome, facilitates the degradation of its contents. LC3B (and its family members) are incorporated into the elongating autophagophore and are necessary for efficient...

Best way to quantitatively measure Autophagic Flux

Tuesday, February 19, 2019 - 10:36
LC3B expression in HeLa cells, ICC, immunocytochemistry, immunofluorescence

By Christina Towers, PhD

Autophagy is a stress-induced cellular recycling process that plays an important physiological role in many diseases. It is induced by a variety of stimuli, both intracellular and extracellular, and is a tightly regulated process with over 30 core autophagy proteins necessary for efficient autophagosome formation and fusion with lysosomes to facilitate degradation1. One of the most common ways to...

Wet or dry protein transfer: Which one to choose for your western blotting?

Tuesday, February 12, 2019 - 10:50
Novus Biologicals western blot resources page

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...

Animal Models to Study Autophagy

Tuesday, February 5, 2019 - 09:03
Autophagy Signaling Pathway, Novus Biologicals, macroautophagy induction, elongation, microautophagy, autophagosome maturation, autolysosome

By Christina Towers, PhD

What is autophagy?

Autophagy is the catabolic process that degrades cytoplasmic material via the lysosome. The process of macroautophagy was originally characterized in yeast, where the core autophagy-related proteins (ATGs) were identified. In this system, the multistep process of phagophore initiation and elongation as well as the conjugation machinery that mediate efficient autophagosome completion were determined and cloned to assess function. The mechanisms of...

Sample collection from mammalian culture cells for kinomic analysis

Tuesday, January 29, 2019 - 10:10
ERK1/2/5 Signaling Interactive Pathway

By Jamshed Arslan Pharm.D., PhD.


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...

Best Methods to Induce and Inhibit Autophagy Pharmacologically

Tuesday, January 22, 2019 - 09:33
VPS35 expression in endosomes and lysosomes of human U-2 OS cells, ICC

By Christina Towers, PhD

Autophagy facilitates the degradation and recycling of damaged cytoplasmic material. The multistep process includes a double membrane structure called an autophagosome that engulfs proteins and organelles and then fuses with lysosomes to facilitate their breakdown. Autophagy is tightly regulated by nutrient availability and is negatively regulated by the robust nutrient sensor mammalian target of rapamycin (mTOR). Starvation, either by glucose deprivation or amino acid depletion, causes a marked increase in autophagy, best quantified by an increase in...


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