Antibody News

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

Monday, July 25, 2016 - 14:48

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).

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FANCD2 and DNA damage repair

Friday, July 22, 2016 - 13:28

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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Tat-Beclin 1: The pioneering of an autophagy-inducing peptide

Wednesday, July 20, 2016 - 11:37

Autophagy is an essential process that maintains cellular homeostasis and carries out lysosome-mediated degradation of unwanted proteins in the cytoplasm.  Because of this regulatory function, autophagy is often examined when looking at disease pathways.  While our immune system initiates the removal of viruses and pathogens through the autophagic pathway, viruses, such as HIV, have developed a way to evade this process through inhibition.  Therefore, developing a reliable way to examine the molecular process of this inhibition and interaction is very desired.  The central autophagy protein, Beclin 1, became a quick target to create an autophagy manipulation factor and ultimately lead to the formation of an autophagy inducing peptide. Novus Biologicals carries three new isoforms of the Tat-beclin-1 peptide in the form of Tat-Beclin 1 D11 (NBP2-49888),...

MHC Class I and the Herpes Simplex Virus

Friday, July 15, 2016 - 14:41

MHC molecules (also known as major histocompatibility complex molecules) assist in the presentation of antigens to T cells in order to eradicate foreign pathogens.  These molecules are highly polymorphic, meaning that they exist in multiple variants in order to avoid pathogens evading their activation of the immune response.  MHC Class I molecules in particular deliver cytosolic peptides to the cell surface so that they can continue on through the cytosol and ultimately the endoplasmic reticulum (ER).  The mechanisms by which different viruses invade our immune system are dynamic and specific to the genetic composition of the virus.  An interesting virus in its process of immune evasion is the Herpes Simplex Virus Infection, given its ability to establish a lifelong cycle of dormant and active phases.  Using a MHC Class I antibody is an effective tool to examine different approaches at virus evasion in the following...

Interactions between CENPF and the additional kinetochore assembly proteins PinX1, PHB2 and Sgt1

Tuesday, July 12, 2016 - 12:58

Mitosis, the process of cell division, involves unique interactions between spindle microtubules and chromosomes, which are regulated by protein structures located on chromosomes known as kinetochores.  CENPF is a kinetochore-associated protein that is localized to chromatin during the G2 and M phases of mitosis.  The main role of CENPF is to secure kinetochore proteins to the correct location surrounding the chromatid and centromere for proper function. Specifically, microtubule-dependent motor proteins work alongside the kinetochores to generate tension and ultimately power chromosomal movement. Outside of its role in cell division, CENPF also has a role in the regulation of the plasma membrane through its association with SNAP25 (a vesicle associated membrane protein).  Research has implicated a wide variety of proteins as regulators of kinetochore and chromatin dynamics; however, this article will review how a...

TIM-3, a critical immune checkpoint in HIV research

Friday, July 8, 2016 - 11:47

CD4+ T-helper cells (Th) are the white blood lymphocytes expressing surface glycoprotein antigen CD4. These T-helper cells play an important role in the adaptive immune system by releasing T cell cytokines that help other immune cells to suppress or regulate immune responses. CD4+ T-helper lymphocytes can be divided into two types (Th1 and Th2) based on their cytokine secretion. Th1 cells are involved in cell-mediated immune response to intracellular pathogens and delayed-type hypersensitivity reactions. Th2 cells are involved in the immune response to extracellular infections and the promotion of allergic diseases.

T-cell immunoglobulin and mucin-domain containing-3 (TIM-3), also known as HAVCR2, is a Th1-specific cell surface protein that serves as an immune checkpoint by inhibiting the immune response. Specifically, TIM-3 regulates macrophage activation, inhibits auto- and...

CD3 (OKT3) as a marker of immune response efficiency

Tuesday, July 5, 2016 - 15:03

Our immune system is a powerful defense mechanism against infection, however different variables can cause our immune response to work for or against us.  CD3 (cluster of differentiation 3) is one component of our immune signal response that is composed of four distinct chains (CD3-g, CD3-e, CD3-s and the zeta chain). These chains associate with a molecule known as the T-cell receptor (TCR) to comprise the TCR complex. Broadly, CD3 is expressed in pro-thymocytes (stem cells where T cells arise in the thymus) in order to mediate signals that are critical for T cell development and function in response to foreign pathogens.

The CD3 (OKT3 Clone) antibody (Cat# NBP2-24867) specifically reacts with an epitope on the epsilon subunit, which plays a vital role in the creation of new T cells.  OKT3 has been found to illustrate immunosuppressive properties and...

Nogo: A Promising Target for New Gene Therapies

Wednesday, June 29, 2016 - 15:31

Nogo is a neurite outgrowth inhibitor protein that plays an important role during central nervous system (CNS) development as well as in endoplasmic reticulum signaling regulation. Studies using Nogo antibodies have revealed Nogo proteins regulate precursor migration, neurite growth and branching in the developing CNS. In addition, Nogo serves as a negative regulator of neuronal growth in the adult CNS, causing wiring stabilization but greatly limiting any regeneration abilities (Schwab, 2010).

There are three Nogo isoforms; Nogo-A, Nogo-B and Nogo-C. Although Nogo-A was the first identified isoform and is the most widely studied, all three isoforms have shown significant potential as targets for a variety of new gene therapies.

A number of recent studies have shown that inhibition using...

The affects of Perilipin 2 on diet and metabolism

Monday, June 27, 2016 - 14:51

Perilipin 2 belongs to the Perilipin family, which consists of proteins that coat intracellular lipid storage droplets. Perilipin 2 in particular is involved in lipid globule surface membrane composition, and has also been implicated in the development and maintenance of adipose tissue. Contrary to previous findings, Perilipin 2 is found in a variety of cells aside from adipocytes, ranging from fibroblasts to skin cells. Overall, perilipins regulate cellular lipid metabolism and homeostasis.  On the contrary, excess lipid stores are linked to both exercise and obese conditions.  Maintaining the correct balance of lipid storage is very important to avoid lipocytotoxicity and is the focus of obese and metabolism related pathologies. 

Perilipin 2 antibody

Perilipin-2/ADFP Antibody [NB110-40877] -...

Synapsin I: Implicated in synaptic activity across a diverse range of studies

Wednesday, June 22, 2016 - 11:29

Synapsins are a family of neuronal proteins that are most renowned for their activity in modulating the pre-synaptic terminal.  Synapsin’s behavior is regulated by protein kinases and phosphatases, which alter the way that synapsin’s interact with actin filaments and other nearby proteins.  There are three isoforms of Synapsin – Synapsin I, II and III.  Synapsin I specifically localizes to the membrane of presynaptic vesicles and plays a role in regulation of axonogenesis and synaptogenesis. Mutations in Synapsin I have been associated with X-linked disorders with primary neuronal degeneration and epilepsy. 

Federici et al used a Synapsin I antibody (Cat# NB300-104) as a biomarker in their study on depression of synaptic dopamine release with cocaine and methylphenidate use.  The Synapsin I antibody was used in western blot to follow the effects of...

FOXP3

Monday, June 20, 2016 - 11:52

Is has been established that the regulatory transcription factor FOXP3 (a member of the forkhead/winged-helix family of transcription factors) is imperative to immune system homeostasis through CD4+CD25+ regulatory T cell function.  Distinctively, FOXP3 binds to specific regions of DNA to modulate the activity of genes that are involved in regulating the immune system.  Interruption of FOXP3 activity leads to autoimmune disorder, due to Treg cells not having their full ability to act as an immune system balancer.

However, it has also been shared that FOXP3 is also expressed and correlated to a number of cancer tissues. This does not come as a surprise, given that FOXP3 regulates a broad spectrum of target genes through histone modifications of target promoters.  The FOXP3 locus itself has CpG dinucleotides that are methylated in naïve CD4/CD25 T cells, activated CD4 T cells, and TGF induced adaptive Tregs.  Now, the FOXP3...

Analysis of Total & pSer724 IRE1 alpha, the Sensor of ER Stress

Friday, June 17, 2016 - 13:01

Inositol-requiring protein 1/IRE1 alpha (also called Endoplasmic Reticulum to Nucleus Signaling 1/ERN1; predicted mol wt 110 kDa) is a serine-threonine protein kinase/endoribonuclease which plays a highly critical role in unfolded protein response/UPR signaling, a mechanism by which eukaryotic cells sense and deal with ER stress. The latter triggers growth arrest and apoptosis in cells with misfolded proteins. ER stress is initiated by three different ER-resident proteins: PERK (PRKR-like ER kinase), ATF-6 (activating transcription factor 6), and IRE1 Alpha, but the signaling mediated by IRE1 Alpha is the most conserved mechanism of UPR/ER stress in eukaryotes. IRE1 Alpha is expressed as a type-1 transmembrane protein in the ER and it has bifunctional cytosolic kinase as well as RNase domains....

The role of TLR4 in breast cancer

Wednesday, June 15, 2016 - 11:40

Toll like receptors (TLRs) are highly conserved proteins that are first known for their role in pathogen recognition and immune response activation.  In order to elicit the necessary immune response in reaction to a foreign pathogen, TLRs trigger cytokine production depending on the behavior patterns of the pathogen itself.  Specifically, TLR4 acts through bacterial lipopolysaccharide (LPS), which composes the outer wall of Gram-negative bacteria.  Bacterial LPS is also a potent activator of the immune system.  Essentially, LPS is a ligand to TLR4, which in turn interacts with myeloid differentiation protein 2 (MD-2), CD14 and LPS-binding protein (LBP), which exist in the extracellular space.  This interaction turns on a signaling cascade that leads to the production of the required cytokines to trigger an effective immune response. 

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Using SCP3/SYCP3 Antibodies as Meiosis Markers in Gametogenesis and DNA Repair Studies

Monday, June 13, 2016 - 11:08

The synaptonemal complex (SC) is a protein structure that forms during the synapsis of homologous chromosomes during meiosis. This structure is involved in the processes of chromosome synapsis, genetic recombination and subsequent chromosome segregation, and is essential for gametogenesis.

The SC is comprised of three component proteins SYCP1, SYCP2, and SYCP3. SYCP1 functions in SC assembly, meiotic recombination, and XY body formation (de Vries et al., 2005). SYCP2 is required for synaptonemal complex assembly and chromosomal synapsis (Yang et al., 2006). Synaptonemal complex protein 3 (SCP3/SYCP3) is an essential structural component of the SC lateral complex (Alsheimer et al., 2010). Mutations in the SYCP3 gene have been linked to azoospermia in males and recurrent pregnancy loss in females (Sazegari et al., 2014), suggesting it plays a critical role in proper gametogenesis.

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Synapsin I, a pre-synaptic marker

Wednesday, June 8, 2016 - 14:40

Synapsin-I, also called Synapsin 1/Syn1, is an ~80 kDa protein (predicted mol. wt. 74.1 kDa) which belongs to the Synapsin family (Synapsin I, Synapsin II, Synapsin III). Synapsins are the evolutionarily conserved phospho-proteins which are associated with the cytosolic side of the synaptic vesicles. They tether the vesicles to the actin cytoskeleton, thus forming a reserve pool. Synapsins I and II are generally found in mature synapses, whereas, Synapsin III is typically expressed in developing synapses with a relatively lower expression. Synapsins represents the most abundant of neuron-specific phospho-proteins, consisting of 9% of the total amount of all vesicle proteins in neurons wherein they play a critical role in the regulation of neurotransmitter release (Greengard et al. 1993)....

The diverse functions of RANKL/TRANCE/TNFSF11

Monday, June 6, 2016 - 11:39

RANKL (also known as TNF-related activation-induced cytokine), or receptor activator of nuclear factor-κB ligand, was first discovered as a key player in the RANKL/RANK/OPG osteoclast formation pathway. Osteoclasts are large multinucleate cells that absorb bone tissue throughout growth and healing cycles.  Osteoclasts express RANKL, which in turn binds to the RANK receptor to regulate osteoclast differentiation.  In healthy bone formation, dynamics shift between new bone formation and existing bone reabsorption, resulting in an important equilibrium that results in osteoporosis or other bone pathologies if disrupted.  Specifically, overstimulation of RANKL activity has been linked to osteoporosis, osteopetrosis, arthritis and additional bone malignancies. 

trance antibody

In the presence of Recombinant Mouse M-CSF (20 ng/mL, Catalog...

KLF4 as a transcription factor in stem cell differentiation

Friday, June 3, 2016 - 12:58

Krüppel-like factors (KLFs) are evolutionarily conserved zinc finger transcription factors that play a role in cell differentiation, proliferation, and pluripotency. KLF4 has specifically been tied to many diverse cellular processes, including self-renewal, apoptosis, and the correct development of the barrier function of skin.  KLF4 can act as both an activator and a repressor, with these functions often being initiated during embryonic development.  Not surprisingly, KLF4 is highly expressed in the skin and gut, and aids in kidney and skeletal development.  Mutations in KLF4 have led to metabolic disorders, cardiovascular disease and many various cancers.  In fact, KLF4 has been as shown through research to have the ability to reprogram adult fibroblasts into induced pluripotent stem cells, and the reprogramming of cancer cells with pluripotency factors has been recently proposed as a potential cancer therapy.

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How Adenovirus and Adeno-Associated Virus Work as Gene Therapy Vectors

Wednesday, June 1, 2016 - 14:38

Adenoviruses comprise a family of medium sized, non-enveloped viruses that were originally isolated from human adenoids (Rowe et al., 1953). These viruses contain a double stranded DNA genome within an icosahedral nucleocapsid capable of penetrating an endosome without the need for envelope fusion. Researchers using adenovirus antibodies have since identified more than fifty human adenovirus serotypes, which are known to cause a variety of diseases; including upper respiratory tract infections, gastroenteritis, conjunctivitis, and cystitis. Several types have even demonstrated oncogenic potential.

Recombinant adenoviruses have long been seen as useful gene therapy vectors (Heubner et al, 1964), due to their ability to encode proteins without integrating into the host cell genome (Roy-Chowdhury et al., 2002). As adenoviruses infect a host cell, their DNA is not incorporated into the cell's...

ABC Membrane transporters and the role of MRP1 in drug resistance

Thursday, May 26, 2016 - 10:39

ATP-binding cassette (ABC) transporters, alongside ion channels and aquaporins, are ubiquitous membrane-bound proteins that move substrates across extra and intra cellular membranes.  Multidrug resistance-associated protein 1 (MRP1) is a member of the superfamily of ATP-binding cassette (ABC) transporters, and in humans is encoded by the ABCC1 gene. In addition to endogenous substrate exchange, ABC transporters also participate in the movement of drugs and chemicals across cell surface and cellular organelle membranes, suggesting that defects in these genes are of great importance in cancer therapy and pharmacokinetics.  In fact, MRP1 was initially identified due to its ability to yield multidrug resistance in lung cancer cells.  MRP1 is one of many genes that may alter a tumor cell to expel cancer drugs and deem them ineffective. Specifically, MRP1 acts as a pump, disposing of cytotoxic chemicals from tumor cells. Drug resistance in chemotherapy is a constant...

Why LC3B Antibodies Make Ideal Autophagosomes Membrane Markers

Tuesday, May 24, 2016 - 15:07

The human form of microtubule-associated protein light chain 3 (LC3) is expressed as 3 splice variants; LC3A, LC3B and LC3C (He et al., 2003). LC3B is a subunit of the MAP1A and MAP1B microtubule-binding proteins and plays a central role autophagosome membrane structure. This ubiquitin-like modifier is known to be involved in early stages of autophagosome formation and specifically with phagophore membrane elongation. LC3B also interacts with autophagy receptors such as p62/SQSTM1 and NBR1 during substrate selection for autophagic degradation. (Johansen et al., 2011).

Researchers using LC3B knock out mice to determine if LC3 is required for autophagy found that the mice do develop normally, likely due to a compensatory autophagy mechanism by other members of the MAP1LC3...

Controls for HIF-1 Alpha's WB, ICC-IF, IHC, IP & FLOW Analysis

Friday, May 20, 2016 - 13:57

Tips on positive and negative controls for HIF-1 alpha antibodies is one of the most Frequently Asked Questions on Hypoxia and HIFs. Here are top 5 suggestions from Novus Biologicals:

  1. The degradation of HIF1 alpha is the most common issue which often results in either a weaker specific signal and/or the appearance of multiple degraded protein bands in Western blot (WB). Degradation may be avoided by preparing the lysates quickly (on ice/at 4˚C) after the collection of cells or tissues, preferably in a hypoxic chamber. For differentiating the target band from degradation (40-80kDa) or dimer (200+ kDa) bands, we suggest including a control based on true hypoxia or hypoxia mimetic samples (e.g. cells treated with CoCl2). Novus offers the following ready to use lysates for HIF-1 analysis.

NFkB and p62 Both Activate and Regulate Inflammation

Thursday, May 19, 2016 - 14:53

Nuclear factor kappa-light-chain-enhancer of activated B cells (NFkB) is a protein complex that regulates DNA transcription and is a critical regulator of cell survival. NFkB has long been known as a primer of inflammation, however researchers are now finding that NFkB may also regulate over-inflammation via a novel mitophagy pathway (Minton, 2016).

NFkB proteins are structurally homologous with retroviral oncoproteins originally known as v-Rel, and are now classified together as NF-κB/Rel family proteins. These include NFkB1 p50/p105, NFkB2 p52/p100, RelA/NFkB p65, RelB and c-Rel, which can function together as dimeric transcription factors to regulate gene expression via the NFkB signaling pathway (Gilmore 2008).

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The TGR5 Receptor and metabolic disease

Monday, May 16, 2016 - 15:16

The bile acid-responsive G-protein-coupled receptor TGR5 is widely distributed across the human body - including the endocrine glands, adipocyte cells, muscles, immune organs, spinal cord, and the enteric nervous system. G protein coupled receptors (GPCRs) are incredibly versatile signaling molecules that are activated by a number of ligands, which in turn regulate various signaling pathways. Select G proteins stimulate activity, whereas others have inhibitory effects.  The effect of TGR5 activation depends on the tissue where it is expressed and the signaling cascade that it induces.  Specifically, bile acids initiate the activation of the TGR5 receptor.  BA’s primary role are to facilitate the uptake and absorption of lipids, making their receptors important for bile acid synthesis and metabolism, as well as integration of multiple homeostatic functions in the liver and gastrointestinal tract.  Since its discovery, TGR5 has been...

Wnt-5a Antibodies Help Understand Wnt Mediated Signaling in Embryogenesis and Various Diseases

Friday, May 13, 2016 - 13:57

Wingless-Type 5A (Wnt-5a) is a member of the WNT family of secreted signaling proteins that regulate many important developmental processes including cell proliferation, migration, differentiation, fate determination and embryonic patterning. WNT signal proteins affect the cell via three known WNT signal transduction pathways. The canonical WNT signaling pathway regulates gene transcription, the non-canonical planar cell polarity pathway regulates cytoskeletal formation, and the non-canonical Wnt/calcium pathway regulates cellular calcium levels.

Wnt5a signals through both the canonical and non-canonical WNT pathways and plays an essential role in regulating developmental pathways during embryogenesis. Mutations in the Wnt5a gene are associated with Robinow syndrome (Roifman et al., 2015) and may also play a role in oncogenesis.

Recently, Wnt5a has been shown to regulate embryonic cell proliferation and differentiation in...

Understanding the relationship between HIF-1 alpha, Hypoxia and Epithelial-Mesenchymal Transition

Tuesday, May 10, 2016 - 14:08

Epithelial-mesenchymal transition (EMT) is a natural process by which epithelial cells lose their polarity and intercellular adhesion, and gain the migratory invasive properties of mesenchymal stem cells that can differentiate into a variety of cell types. EMT is critical to many developmental processes including embryo development and wound healing. However, EMT is also a fundamental step in the initiation of metastasis during cancer progression.

Hypoxia is known to induce EMT via HIF-1 alpha induction. Researchers have used HIF-1 alpha antibodies (NB100-105), (NB100-449) and (NB100-479) to help identify the molecular mechanisms...

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