Antibody News

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...

The human form of microtubule-associated protein light chain 3 (LC3) is expressed as 3 splice variants LC3A, LC3B, and LC3C.¹ LC3B is a subunit of the MAP1A and MAP1B microtubule-binding proteins and plays a central role in 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.²

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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).

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...

DOPA decarboxylase (DDC) is responsible for catalyzing the conversion of aromatic amino acids into their corresponding amines during the synthesis of several important neurotransmitters. Specifically, DDC catalyzes the decarboxylation of L-DOPA to dopamine, L5-HTP to serotonin, L-histidine to histamine, phenylalanine to phenethylamine, L-tyrosine to tyramine, and tryptophan to tryptamine.

A number of mutations in the DDC gene can cause aromatic L-amino-acid decarboxylase deficiency (AADCD) (Gücüyener et al., 2014). AADCD is an inborn error in neurotransmitter metabolism that leads to combined serotonin and catecholamine deficiency. DDC's critical role in neurotransmitter synthesis makes it an important target in the study of Parkinson's disease, depression and other neuroscience related diseases.

Parkinson's patients experience motor system symptoms due to reduced levels of dopamine in the substantia nigra portion of the brain. The conversion of L-DOPA...

EZH1 is part of the Polycomb-group family of proteins, which are responsible for remodeling chromatin in genes and modulating epigenetic silencing during development.  Specifically, EZHI is a component of PRC2, or polycomb repressive complex-2.  PRC2 interacts and modifies the histone “H3”, and is critical in maintaining gene repression.  EZH1 has the ability to mono-, di- and tri-methylate 'Lys-27' of histone H3 to form H3K27me1, H3K27me2 and H3K27me3, respectively.  EZH1 is also implicated in the maintenance of embryonic stem cells, including their differentiation and renewal.    

Margueron et al took a closer look at the mammalian homologs of EZH1 and EZH2 in order to learn more about their similarities and differences during development (1). They found that while both Ez proteins form PRC2 complexes, they carry out different repressive roles.  Ultimately, both protein complexes are efficient at...

PARPs (poly ADP ribose polymerases) are DNA repair enzymes that promote single stranded break (SSB) repair by binding to DNA at the sites of SSBs and recruiting repair machinery. In humans, the PARP superfamily consists of 17 members, of which five play known roles in SSB repair. PARP-1, the most well-studied family member, is required for base excision repair and is thought to be responsible for 90% of PARP activity (5).

PARP inhibitors are a class of pharmacological agents that have been used to treat certain types of cancer, and are designed to induce cell death by preventing cancer cells from repairing damaged DNA. PARP inhibitors have been tested in various combination therapy contexts. For instance, one recent study compared radiation-induced cytotoxicity in cell culture and xenograft models of pancreatic carcinoma and found that PARP inhibition with radiation increased cell death in vitro and reduced tumor...

HIF-2 alpha is a member of the heterodimeric hypoxia-inducible factors/HIFs family (HIF-1, HIF-2, and HIF-3) which contains a common beta subunit but differ in their alpha subunits. Also called as EPAS1 or Mop2, HIF-2 alpha regulates cellular adaptation to hypoxia which is involved in several biological processes such as angiogenesis, cellular survival/proliferation, energy metabolism, erythropoiesis, extra-cellular matrix functions, invasion/ metastasis, iron metabolism, pH regulation, multidrug resistance, stem cell properties. The expression of HIF-2 alpha is regulated through hypoxia-dependent protein stabilization with the help of proteins

What are Caspases?

Caspases, or cysteine-dependent aspartate specific proteases, are a family of enzymes crucial for initiating and executing apoptosis within a cell, an important biological event especially during organ development (1). Environmental cues and cellular signals trigger the initiation of the programmed cell death cascade primarily through proteolytic activation of the caspases (1). One specific effector caspase is caspase-3, a protein that is cleaved and thus activated upon the initiation of apoptosis. Cleaved caspase-3 propagates an apoptotic signal through enzymatic activity on downstream targets, including poly ADP ribose polymerase (PARP) and other substrates (2). In cell biology, caspase-3 antibodies that detect both uncleaved and cleaved versions of the enzyme are strong indicators...

Novus Biologicals' antibodies are the gold standard to monitor autophagy and detect LC3 expression. The recently published Guidelines for the Use and Interpretation of Assays for Monitoring Autophagy (3rd Edition) comprehensively details methods to monitor autophagy in cell or tissue samples. Importantly, these guidelines also provide key considerations for data interpretation and tips to creating better western blot data.  

Use fresh samples and avoid multiple freeze thaws to improve detection of LC3-I when possible.

Why? LC3-I is less stable and more susceptible to freeze thaws and degradation in SDS compared to LC3-II. Moreover, most LC3 antibodies are less sensitive to LC3-I...

GFAP, a class-III intermediate filament, is a 50kDa protein which is found in the mature and developing astrocytes in the CNS, non-myelinating Schwann cells in the PNS, enteric glial cells (enteric nervous system/ENS), ependymal cells, and radial glia of the developing brain. GFAP antibodies are the most popular marker for astrocytes in neurological studies and along with its breakdown products (BDPs), GFAP has been proposed as a useful candidates for biofluid-based markers for numerous neurological conditions especially during traumatic brain/spinal cord injury and stroke [1]. Moreover, neuro-oncology studies have established a positive reaction to GFAP in astrocytomas, ependymoma, and astrocytic cells of mixed gliomas, subependymal giant cell astrocytoma, pleomorphic xanthoastrocytoma, astroblastoma, and gliosarcoma [2].

Western blotting is one of the most commonly used antibody assay techniques in cell and molecular biology research since its development over three decades ago, and is considered the gold standard for protein detection and quantification.

When performing a Western Blot, it is crucial to ensure equal loading of protein samples and protein transfer through the use of a loading control antibody. beta-Actin, GAPDH and alpha Tubulin are well known housekeeping proteins that are commonly used as loading controls. While there are various options available, researchers often face some challenges in choosing a proper loading control antibody as expression of housekeeping proteins can be influenced by sample type, treatment or other...