Antibody News

Using a STAT3 antibody in chromatin immunoprecipitation (ChIP)

Friday, October 21, 2016 - 14:38

Signal transducer and activator of transcription 3 (STAT3) is an important oncogenic transcriptional factor that mediates tumor induced immune suppression.  Specifically, STAT3 transmits signals from cytokines and growth factor receptors in the plasma membrane (PM) to the nucleus, where they alter gene transcription.  Because of this transcriptional regulatory role, STAT3 also plays a part in regulating transcription of many critical genes that are involved in apoptosis, cell differentiation, immune response, tumor formation and more.  Using a STAT3 antibody in the application of chromatin immunoprecipitation (ChIP) is a very effective way to examine the relationship between genes and proteins in different molecular pathways and disease pathologies.  The next two studies used a...

Beta Amyloid (MOAB2) and the link between traumatic brain injury and Alzheimer’s disease

Tuesday, October 18, 2016 - 13:25

An epidemiological association between traumatic brain injury (TBI) and Alzheimer's disease (AD) has long been established.  Interestingly, an increase in beta amyloid  (one hallmark of AD) directly following TBI has been observed.  In fact, it has been reported that with a greater level of TBI comes a higher risk of developing AD, or other neurodegenerative disorders, in the future.  Roberts et al first presented research that beta amyloid plaques found in TBI patients are very similar to those found in AD patients. Amyloid precursor protein (APP) is a transmembrane protein that has high expression levels in the brain.  In the nonamyloidogenic pathway, APP is cleaved by gamma and beta secretases, which can produce amyloid beta fragments.  These amyloid beta fragments become misbalanced in the brain, which contributes to how amyloid beta is produced and regulated.  In order to better...

Winter is coming, and TRPM8 welcomes the cold!

Friday, October 14, 2016 - 13:57

TRPM8, or transient receptor potential melastatin 8, is a nonselective cation channel that is activated by cold environments and menthol-like cooling compounds.  While TRPM8 is best known for its location in peripheral nerve endings, it has functionality both inside and out of the nervous system.  Within the nervous system, TRPM8 is responsible for our response to cold and or menthol like stimuli.  Our reaction to cold sensation is involved in a variety of processes and can be a part of reactions such as asthma.  Outside of the nervous system, TRPM8 has shown high expression in prostate cancer tumors and is thought to potentially play a role in prostate cancer cell migration.  The TRPM8 protein is composed of six transmembrane domains with a transient receptor potential (TRP) domain at the C-terminal end.  In addition to cold temperatures (below 26 degrees Celsius) and menthol like compounds, antagonists such as capsazepine (synthetic analog of capsaicin, a heat...

Five Tips to Successful Western Blot of phospho-IRE1 alpha (Ser724)

Friday, October 7, 2016 - 13:20
1. Sample Preparation

Phospho-IRE1 alpha (Ser724) antibodies will detect IRE1 alpha protein only when it is phosphorylated at the Serine-724 amino acid position. If IRE1 alpha is not phosphorylated (activated) in the samples being tested, phosphorylation specific IRE1 alpha antibodies would not generate a signal. Therefore, it is recommended to use a positive control from cells with high amounts of ER stress/UPR activation. Our in-house testing for phospho-IRE1 alpha (Ser724) expression with Novus’ phospho-IRE1 alpha (Ser724) antibody (NB100-2323), has demonstrated that Min6 cells exposed to increasing concentrations of glucose (up to 20nM for 3 hours) or HeLa cells exposed to Dithiothreitol/DTT (10 mM for 1 hour) serve as great options for positive controls in phospho-IRE1 alpha expression analysis through Western blot....

Thomson Reuters Predicts 2016 Nobel Prize Winners

Friday, September 30, 2016 - 07:36

Here at Bio-Techne we always look forward to the annual announcements of winners of the highly coveted Nobel Prize – the greatest award in science. How can you go about predicting which scientists might be in line for a life-changing phone-call from the Nobel Committee?

Well, looking at citations is one possible approach.

Thomson Reuters analysts annually mine scientific citation data to identify the researchers whose work is worthy of Nobel recognition, and are responsible for the world's most influential scientific innovations. These are the scholars and innovators, whose papers typically rank in the top 0.1% by citations within their field. Many of these go on to win the Nobel Prize for their contributions towards the advancement of science.

The list of 2016 Citation Laureates has been released, prior to the announcement of Nobel Prize winners, beginning on 3 October. Thomson Reuters has had a degree of success in predicting which scientists...

Meeting Report: 2nd International Antibody Validation Meeting

Tuesday, September 27, 2016 - 12:54

Bio-Techne brands Novus Biologicals® and R&D Systems® were proud to support the 2nd International Antibody Validation Meeting held at Bath University, on the 15-16 September, 2016. Almost 100 participants from around the world, including funders, publishers, academics, pharma and antibody manufacturers were in attendance, to share and discuss best practices in research antibody validation and to drive improved antibody validation standards for the global life science community.

Biotechne booth

Bio-Techne booth

Release of highly validated antibodies, and research reproducibility is an issue which we at Bio-Techne have taken seriously for the last 30 years. The meeting was a timely and ideal forum to share our decades of antibody development and validation experience, and stringent quality control criteria, with the rest...

Altered expression of BCL2 in cancer

Friday, September 23, 2016 - 09:16

Similar to other cell processes, the balance between cell survival and cell death is an important equilibrium that when altered expression of genes can lead to a variety of disease.  For example, too little cell death can promote cell overgrowth and eventually cancer, whereas too much cell death can lead to neurodegeneration (among other things).  Bcl-2 in particular is a pro-survival protein that is part of the Bcl-2 family of proteins, consisting of Bax, Bid, PUMA, and Noxa.  While overexpression of Bcl-2 has rescued cells from certain toxic stimuli, such as hypoxia, of greater interest is its ability to prevent cell death during a chemotherapeutic treatment and its resistance to these drugs.  In fact, an upregulation of Bcl-2 family member proteins has been used as a light prognostic factor in diagnosing certain cancers.  Given this information, the use of Bcl-2 primary antibodies in research has been successful in elucidating...

The effect of antioxidants and the NFkB p65 pathway in inflammation

Tuesday, September 20, 2016 - 13:55

NFkB is a transcription factor that plays a role in the expression of genes involved in immune response, inflammation, metastasis, cell survival and more. RelA (p65) is one member of the NFkB mammalian family, alongside other subunits. NFkB subunits have recognition sites on the “Rel” homology domain, where they form protein complexes to bind DNA and regulate gene expression.  The RelA (p65) subunit also has an extended carboxy terminal that can act as a transactivator (a form of gene regulation that increases the rate of gene expression in response to stimuli).  The NFkB p65 subunit is thought to shuttle between the cytoplasm and the nucleus, however it is also shown to stay localized to the cytoplasm by its inhibitor, IKK beta.  Over the past ten years or so, introduction of antioxidants to inflammation or aging has had interesting response on a cellular level.  Given that the NFkB pathway regulates the levels of endogenous reactive oxygen species (ROS),...

Alpha-smooth muscle actin and the modulation of endothelial and epithelial cell biology

Friday, September 16, 2016 - 14:36

Actin is essential for a wide range of cell functions, ranging from cell division and chromatin remodeling to vesicle trafficking and maintenance of cellular structure. In fact, mislocalization of actin to cell junctions during development leads to facial malformations such as cleft lip.  Actin is successful in the regulation of a variety of cell functions due to its high number of isoforms.  One such isoform of actin is alpha-smooth muscle actin (alpha-SMA), which is plentiful in vascular smooth-muscle cells and plays an important role in fibrogenesis and fibrosis (the thickening of tissue). There are distinct phenotypes associated with a lack of alpha-SMA during and after development.  Studies have shown that mutated alpha-SMA during development results in disarray of cardiac muscles and intense muscle weakness in young rodent infants.  In adults, mutations in smooth muscle actin can lead to cardiac and blood pressure complications. 


Beta Tubulin III and neurogenesis

Thursday, September 15, 2016 - 13:32

Beta tubulin III, also known as Tuj-1, is a class III member of the beta tubulin protein family. Beta tubulins are one of two structural components that form our microtubule network. While general tubulins play a role in a wide range of cellular processes (mitosis, motility, etc) beta tubulin III is specifically localized to neurons.  In particular, beta III tubulin’s expression correlates with the earliest phases of neuronal differentiation.  For this reason, beta tubulin III has implications in neurogenesis, axon guidance and maintenance.  Since it was discovered that the human brain generates new neurons from stem cell pools, beta tubulin III has been used as a marker of positive neuronal activity in many research studies.  The following articles use a beta III tubulin antibody in their research of embryonic stem cell and neural progenitor activity. 


MMP-2: More Than a Cancer Marker

Friday, September 9, 2016 - 14:36

Matrix metalloproteinases (MMP) are a family of endopeptidases involved in the breakdown of extracellular matrix (ECM) during both normal physiological and disease processes. MMP-2 is a zinc-dependent family member that selectively cleaves collagen and elastin, major structural components of the basement membrane. In addition, MMP-2 has been found to affect a number of non-matrix proteins such as big endothelin-1 (Fernandez-Patron et al., 1999), KISS (Takino et al., 2003), GSK3B (Kandasamy et al., 2009), and CHUK (Olivotto et al., 2013). This diverse activity has led MMP-2 to be linked to a wide variety of physiological processes.

While most MMP members must be proteolytically activated after secretion, MMP2 may be activated while still on the membrane. Specifically, pro-MMP-2 can be activated either extracellularly by proteases, or intracellularly via S-Glutathionylation (Okamoto et al., 2001). Researchers have speculated that MMP-2...

Transferrin and the blood brain barrier

Wednesday, September 7, 2016 - 14:52

Transferrin, an iron binding protein that facilitates iron uptake in cells, is an integral part of a mechanism that may introduce antibody therapies to the central nervous system. Currently, the brain’s ability to take in antibody therapies is limited by the presence of the blood brain barrier. While novel mechanisms to manipulate the transfer of important proteins and antibodies across the BBB have been tested, endogenous receptor-mediated transcytosis (RMT) is one mechanism of particular interest.  Iron transfer is achieved when transferrin bound iron connects with the transferrin receptor, and iron subsequently enters the cell by way of clathrin-mediated endocytosis.  Once inside the cell, transferrin is trafficked to early endosomes, where the iron is dropped off.  After iron delivery, transferrin is directed to recycling endosomes to be brought back to the cell surface.  Using this pathway to deliver therapeutic antibodies across the BBB has shown both...

The relationship between Ki67 and HIF-1 in cancer

Friday, September 2, 2016 - 13:24

Ki67, also known as MKI67, is best known as the leading marker of cellular proliferation. Ki67 is regulated by a balance between synthesis and degradation, and often carries a very short half-life.  First discovered to be located to dividing cells, Ki67 has since been specifically localized to the G1, S, G2 and M phases of mitosis. Soon after, it was discovered that there was a high correlation of Ki67 alongside the p53 (tumor suppressing protein 53), suggesting an implication in cancer. What’s more, the expression of Ki67 is higher in malignant cells versus control cells. With this data, researchers are taking a closer look between the behaviors of Ki67 alongside proliferation in cancer cells.  In particular, the following articles use the Ki67 primary antibody when investigating the role of HIF-1 in different types of cancers.  Hif-1, or hypoxia inducing factor 1, controls our adaptive response...

Tools for Isolation, Quantification and Analysis of Exosomes

Tuesday, August 30, 2016 - 16:00

Exosomes are spherical to cup-shaped bilayered membrane enclosed nanosize vesicles (30-100 nm) which have the ability to shuttle active cargoes between cells. Johnstone et al. 1987 pioneered in documenting the generation of exosomes in differentiating reticulocytes as a result of the fusion of multi-vesicular endosomes/MVBs with the plasma membrane. However, later studies established that exosomes are actively secreted by almost all types of cells through the process of exocytosis and that exosomes can be found in most of the body fluids (blood, saliva, semen and urine). Structurally, exosomes contain various types of biomolecules (lipids, proteins, DNA, RNA etc.) from the parent cells, and the bacterial/viral proteins or nuclear acids in infected cells. Exosomes act as signalosomes and facilitate intercellular communication through delivering bioactive molecules (Tran et al. 2015)


TRIF/TICAM1 and mitochondrial dynamics in the innate immune response

Friday, August 26, 2016 - 13:10

TRIF, also known as toll like receptor adaptor molecule 1 or TICAM1, is known for its role in invading foreign pathogens as part of our innate immune response. TRIF/TICAM1 is a TIR-domain adaptor protein (toll/interleukin-1 receptor) that interacts with the Toll-like receptors (TLRs) through intracellular signaling and recognition of its TIR site.  TLRs are expressed on a variety of cell types, including macrophages, mast cells, endothelial cells, and more.  In addition to TRIF/TICAM1, another universal adaptor for TLRs is myeloid differentiation factor-88 (MyD88).  The activation of both MyD88 and TRIF/TICAM1 results in subsequent activation of the nuclear factor kappa beta pathway (NF-κB). The NF-κB then influences a large range of biological processes, including immunity, inflammation and stress response.  One way that TRIF/TICAM1 defends a host from foreign pathogens is to initiate autophagy or apoptosis in order to clear the intracellular space from these particles...

The identification of dopaminergic neurons using Tyrosine Hydroxylase in Parkinson's research and LRRK2

Thursday, August 25, 2016 - 08:49

Tyrosine hydroxylase (TH) is a crucial enzyme involved in the biosynthesis of dopamine, norepinephrine and epinephrine in the brain.  Specifically, TH catalyzes the conversion of l-tyrosine to l-dihydroxyphenylalanine (l-dopa).  The importance of tyrosine hydroxylase was established early on in when Zhou et al found that TH deficient mice had a lethal phenotype.  While TH is vital to neurotransmitter and neural hormone development, mutations in TH are not solely responsible for Parkinson’s disease (PD).  In fact, mutations in the LRRK2 gene and marked loss of dopaminergic neurons in the substantia nigra are the hallmark signs of PD. However, mutations in TH lead to dystonia DOPA-responsive autosomal recessive disorder, also known as autosomal recessive Segawa syndrome. Using a TH antibody in Parkinson’s research is an effective and popular way to monitor...

Epithelial-Mesenchymal Transition (EMT) Markers

Thursday, August 18, 2016 - 14:15

Epithelial-Mesenchymal Transition (EMT) is the trans-differentiation of stationary epithelial cells into motile mesenchymal cells. During EMT, epithelial cells lose their junctions and apical-basal polarity, reorganize their cytoskeleton, undergo a change in the signaling cascade that defines cell shape and reprograms gene expression. Collectively, these changes increase the motility of individual cells and enables the development of an invasive phenotype. At the molecular signaling level, EMT is regulated through several pathways which are triggered by TGF-beta, HGF, EGF, FGF, VEGF, Wnt, SHH, IL6, HIF1 alpha, and other proteins. At the transcription level, SNAI1/Snail, ZEB1/ZEB2 and basic helix-loop-helix transcription factors (bHLH) drive EMT progression. Accumulating evidence has established involvement of EMT in several biological processes including (but not limited to) embryonic and post-embryonic development, tissue regeneration/wound healing, stem cell...

The role of Smoothened in pulmonary pathologies

Monday, August 15, 2016 - 14:24

The Hedgehog (Hh) family of secreted proteins is involved in a number of developmental processes, one of which is the development of cancer. Past data suggests that the Sonic hedgehog (Shh) receptor is composed of two transmembrane proteins, Patched and Smoothened.  The Hedgehog (Hh) signaling pathway is vital to the development of many tissues during embryogenesis, however, it also has an important role after development.  After development, Hh signaling regulates stem cells and their regenerative function.  When the Hh pathway is awry, signaling may turn oncogenic in nature. This has been witnessed in interactions between Patched and Smoothened, where Patch inhibits the function of Smoothened.  When normal Smoothened activity is restricted or mutated, it can cause unregulated activation of the Hh pathway which leads to cancer.  This post will delve into research using a Smoothened antibody to further elucidate its role...

The recent relationship of BRCA1 and 53BP1

Friday, August 12, 2016 - 11:16

The p53-binding protein 1 (53BP1) is a DNA damage response factor, which is recruited to nuclear structures at the site of DNA damage.  DNA double-strand breaks (DSBs) are mutations that are detrimental to cell viability and genome stability, and must be repaired either through homologous recombination (HR) or non-homologous end joining (NHEJ). 53BP1 specifically promotes both NHEJ as well as the inhibition of HR repair, yet the decision making on a molecular level between these two routes not clearly understood. Recently, a focus has been placed on 53BP1 and the breast cancer gene BRCA1, given that BRCA1 is also an important mediator of our DNA damage response, partially by antagonizing 53BP1 dependent NHEJ.  The following studies take a closer look at the relationship with 53BP1 and BRCA1

Zhang et al introduced the hypothesis that a cell cycle dependent BRCA1-...

The effects of ethanol consumption on glutamate production and xCT

Monday, August 8, 2016 - 14:03

xCT is a sodium independent glutamate transporter that regulates the exchange of extracellular l-cystine and intracellular l-glutamate across the plasma membrane. This process is critical to glutathione production and protection from subsequent oxidative stress. Aside from its standard function, xCT participates in a variety of central nervous system (CNS) functions, including formation of the blood–brain barrier, involvement in drug addiction pathways, neurodegeneration caused by lack of oxidative stress protection, and more.  xCT is primarily localized to neurons and glia in the CNS, which is considering its role in glutathione production and efficacy, given that there are an abundance of enzymes and metabolites that can generate reactive oxygen species in these areas.

xct antibody

xCT Antibody...

The role of DNMT3B in the co-incidence of methyltransferase and tumor suppressor expression in malignancies

Thursday, August 4, 2016 - 15:18

Epigenetics is the process of heritable change in gene activity despite alteration of the hosts DNA sequence, essentially causing a change in a phenotype without a change in the genotype of a host. To change the gene sequence without interfering with the DNA is accomplished by histone and DNA methylation.  Gene silencing in DNA methylation is carried out by DNA methyltransferases 1, 2 and 3a/b (DNMT1, DNMT2, DNMT3A/B). On a broad level, DNMT’s methylate the fifth carbon of cytosine residues in DNA within CG dinucleotides. However, DNMT3B is required for genome-wide de novo methylation and is during development.  DNMT3B also specifically regulates DNA of nucleosomal DNA, however it can also act as a transcriptional co-repressor by associating with CBX4 in lieu of methylation. Studies on the structure of DNMT3 have revealed that DNMT3A and DNMT3B are highly similar in that they both contain a PWWP domain, a PHD-...

The role of DNMT3A in development

Tuesday, August 2, 2016 - 14:56

Epigenetics is the study of heritable change in gene activity despite alteration of the hosts DNA sequence.  Change in gene activity done independently of the DNA sequence is achieved by way of histone and DNA methylation.  Gene silencing in DNA methylation is carried out by DNA methyltransferases 1, 2 and 3a/b (DNMT1, DNMT2, DNMT3A/B). On a broad level, DNMTs methylate the fifth carbon of cytosine residues in DNA within CG dinucleotides. However, DNMT3 on its own is a de novo methyltransferase required for the establishment of genomic methylation patterns during development and reproduction. Studies on the structure of DNMT3 have revealed that DNMT3A and DNMT3B are highly similar in that they both contain a PWWP domain, a PHD-like ADD domain and a catalytic domain. What’s more, the catalytic activities of DNMT3A and DNMT3B have been shown to be attracted to...

Niemann Pick-C1 and cholesterol dynamics

Friday, July 29, 2016 - 09:17

Niemann-Pick type C1 (NPC1) mediates low-density cholesterol transport from late endosomes and lysosomes to other areas of the cell via receptor mediation endocytosis.  Although cholesterol moves freely inside the cell, it cannot independently export out of the lysosome, which is where NPC1 steps in. After NPC1 interacts with lipoproteins and removes or delivers them to their destined compartments, they are hydrolyzed and released as free cholesterol.  Mutations in the NPC1 gene cause Niemann-Pick type C disease, a rare autosomal recessive neurodegenerative disorder characterized by over accumulation of cholesterol in lysosomes and is characterized by both neural and liver degeneration.

Niemann Pick C1 antibody

Niemann-Pick C1 Antibody [NB400-148] - Staining of human...

Application Highlight: Recent uses of TERF2 in immunofluorescence (IF)

Monday, July 25, 2016 - 14:49

Telomeres are a region of repeat nucleotide sequences located at the end of chromosomes to protect our DNA from becoming damaged via end-to-end fusion.  TERF2, or telomeric-repeat binding factor 2, is important for telomere integrity and aids in the formation of the telosome, the telomeric loop, and control of the amount of DNA needed for telomere replication.  When TERF2 is down regulated, an induction of apoptosis and senescence is also witnessed.  TERF2 also interacts with an abundance of proteins in all of the above pathways, including RAP1, Apollo, TOP1, TOP2A and TOP2B.  The visualization of telomeres on chromosomal structures through immunofluorescence (IF) is an important diagnostic tool and is examined more closely in the following research projects using a Novus Biologicals TRF-2 Antibody (NB100-56506).


FANCD2 and DNA damage repair

Friday, July 22, 2016 - 13:30

Fanconi anemia (FA) is a genetically inherited disorder that yields cytogenetic instability, hypersensitivity to DNA crosslinking compounds and defective DNA repair. A variety of genes have been identified within the FA pathway that are referred to as the Fanconi anemia complementation group.  One member of this group, FANCD2, is monoubiquitinated in response to DNA damage.  At this point, FANCD2 specifically localizes to the nucleus to represent the site of DNA repair, often times to the DNA replication fork. In addition to monoubiquitination, FANCD2 can also be phosphorylated by the cell checkpoint kinases ATM and ATR.  Using a FANCD2 antibody to further elucidate the role of FANCD2 the DNA damage pathway is a popular approach to understanding DNA repair in a variety of experimental models.  


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