Antibody News

Detecting HIF alpha and beyond: Best controls for hypoxia Western blot analysis

Monday, October 14, 2019 - 09:17
Analysis of HIF-1 alpha expression in whole cell lysates from HeLa cells in culture treated or untreated with cobalt chloride and independent antibody validation where two different antibodies to HIF-1 alpha were used for immunoprecipitation and detection.

By Rosa Moreno, PhD.

Detecting HIF alpha and beyond: Best controls for hypoxia Western blot analysis

Physiological low levels of oxygen induce normal hypoxic events across biological systems. This hypoxic state activates hypoxia inducible factors (HIFs) to regulate transcription by binding to the hypoxia response element (HRE) region of...

Neurovascular signaling for repair enhances brain metastasis

Tuesday, October 8, 2019 - 09:02
High expression of GFAP (green) identifies astrocytes in rat cerebellum

By Jamshed Arslan, Pharm. D., PhD.

Stroke    is a leading cause of mortality and morbidity worldwide. Cellular players – neurons, astrocytes, endothelial and stromal cells – involved in post-stroke repair through angiogenesis...

Glypican 3 as a biomarker for gastro-esophageal adenocarcinoma

Tuesday, October 1, 2019 - 10:17
Expression of CD81 was detected by immunofluorescence with a mouse monoclonal antibody followed by secondary antibody labeled with DyLight488 and Phalloidin 568 staining for actin visualization

By Jamshed Arslan, Pharm. D., PhD.

Gastroesophageal adenocarcinoma originates from the glandular epithelium of the esophagus, gastroesophageal junction and stomach. The incidence of gastroesophageal adenocarcinoma is declining, but it is still one of the deadliest cancers worldwide. A key reason behind high mortality is that by the time gastroesophageal adenocarcinoma is diagnosed, the cancer has often already metastasized. The search for novel biomarkers underlying gastroesophageal...

Isolating immune cells from peripheral blood: How and when to use density gradient centrifugation

Tuesday, September 24, 2019 - 09:05
Adaptive immunity is mediated by lymphocytes (e.g., T cells and B cells) while innate immunity responses are mediated by several cellular mediators (e.g., Neutrophils, Natural Killer Cells and Eosinophils)

By Victoria Osinski

Lymphocytes, monocytes, and granulocytes, oh my!

There are many different immune cells (leukocytes) that are found in circulation along with other cellular components such as platelets and red blood cells. Red blood cells (RBCs) are the most abundant, followed by neutrophils and lymphocytes. The immune cells can be categorized as either mononuclear cells (monocytes and lymphocytes...

Suppressing breast cancer metastasis: The role of hypoxia-induced RhoB expression and activation

Tuesday, September 17, 2019 - 08:16
Expression of RhoA, B and C detected in the cytoplasm of A431 cells, a model of human epidermoid carcinoma, compared to negative control analyzed in the absence of RhoA Antibody (1A11-4G10) [NBP2-22528]

By Jamshed Arslan, Pharm. D., PhD.

The Ras homologous

Tips to Optimize your Western blot for phosphorylated protein detection

Tuesday, September 10, 2019 - 10:48
Western blotting allows identifying and quantifying proteins from cell lysates for determination of protein expression, sub-cellular localization, post-translational modifications, protein processing, and protein-protein interactions.

By Jamshed Arslan, Pharm. D., PhD.

Protein phosphorylation refers to a reversible post-translational modification in which a protein kinase adds a phosphate group to an amino acid residue of a target protein. Protein phosphorylation, especially tyrosine phosphorylation, is one of the early events in signal transduction in all eukaryotic cells.

Once a cell is lysed, proteases and phosphatases are released that can degrade or...

The Ins and Outs of Survivin

Tuesday, September 3, 2019 - 08:40
Exosomes arise from the late endosomal compartment through inward budding of multivesicular bodies and may contain multiple proteins, lipids, DNA, RNAs and biomolecules of viruses/prions.

By Rachel M.A. Linger, Ph.D.

What is survivin?

Survivin is a small (16.5 kDa) protein normally found in human fetal tissue. In contrast, survivin is typically undetectable in most normal adult tissues. Expression of survivin occurs in several subcellular locations including the cytoplasm, nucleus,...

Explore IHC basics with this new guide: Immunohistochemistry Essential Elements and Beyond

Tuesday, August 27, 2019 - 09:07
3-color fluorescence IHC, nuclei-blue, cytoplasm-red and cytoskeleton-green, Immunohistochemistry Essential Elements and Beyond, Springer book

By Rosa Moreno, PhD.

This new introductory guide to methods and techniques in immunohistochemistry is written by Dr. Alexander E. Kalyuzhny, Bio-Techne, for the Springer series: Techniques in Life Science and Biomedicine for the Non-Expert   . This guide provides a comprehensive introduction along with conceptual and methodological basics for new users to ensure a clear understanding upon which researchers may build their expertise in ...

Chromatin reader domains of DNMT-targeting protein, UHRF1, are responsible for cancerous DNA hypermethylation

Tuesday, August 20, 2019 - 08:58
Expression of DNMT2 in human colon tissue shows cytoplasmic and nuclear staining in glandular cells, IHC

By Jamshed Arslan, Pharm. D., PhD.

DNA methylation represses transcription of many genes, including tumor suppressor genes. A protein called UHRF1 recruits DNA methyltransferases (DNMTs) to establish and maintain DNA methylation. UHRF1 has several domains, most notably:

  • N-terminal ubiquitin-like (UBL) domain
  • TTD (tandem Tudor domain) and PHD (ubiquitin-like, containing plant homeodomain) domains that...

Role of GFAP in astrocytes: Lessons from induced pluripotent stem cells in Alexander disease patients

Tuesday, August 13, 2019 - 10:52
GFAP-green and Neurofilament-red expression in astrocytes of mouse brain cerebellum, IHC

By Michalina Hanzel, PhD

Alexander disease is a progressive and fatal neurological disease with phenotypes ranging from myelination abnormalities, gait ataxia and megalencephaly to predisposition to seizures. It is an autosomal dominant disease with mutations in the glial fibrillary acidic protein (GFAP) gene that result in astrocytic cytoplasmic inclusions, termed Rosenthal fibers, which contribute to the global neurological...

Considerations for Quantitative Western blotting

Tuesday, August 6, 2019 - 09:11
GAPDH expression in human nucleus pulposus cell lysate, WB

By Jamshed Arslan, Pharm. D., PhD.

Since its inception in 1979, Western blotting has undergone several developments. The use of radioactive probes was common throughout 1980s, but utilizing secondary antibodies labelled with fluorescent/chemiluminescent probes has virtually removed the need for radioactivity. Western blotting has become a routine quantitative method in biological research and immunodiagnostics. The following primer discusses some considerations when quantitating Western blot data.


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Antibody treatment can generate microglia-like cells from bone marrow

Tuesday, July 30, 2019 - 10:18
Sox2 antibody expression in induced pluripotent stem cells, ICC

By Jennifer Sokolowski, MD, PhD.

Microglia play important roles in the brain in both homeostatic and pathological conditions, acting to clear debris and dying cells. There is evidence to suggest that microglial dysfunction contributes to neurodegenerative diseases such as Alzheimer's and modulation of microglial activity may be a method to treat such diseases.1 Cell-based therapies represent a novel approach whereby the introduction of microglia that possess the desired phenotype could potentially be...

Optogenetic Control of Mitophagy: AMBRA1 based mitophagy switch

Tuesday, July 23, 2019 - 10:38
sAmbra1 expression and localization to mitochondria in human U-2 OS cell line, ICC

By Christina Towers, PhD

Mitophagy in the Brain

Selective autophagic degradation of damaged mitochondria, known as mitophagy, has been described as a cyto-protective process. Accordingly, defects in mitophagy have been associated with a number of diseases including muscle atrophy, cancer, and multiple neurodegenerative diseases. Defective mitophagy has been best described in neurons, where the accumulation of damaged mitochondria and the resulting increase in...

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 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 inflammatory...

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,...

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

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 Mechanisms of Neurodegeneration: Protein aggregation and failure of autophagy, and the role of mitochondria and...

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
TOMM20 expression in human cerebral cortex mitochondria, IHC

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: Protein aggregation and failure of autophagy.

Reactive...

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...

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...

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