Antibody News

EZH1 has more to offer than gene

Friday, April 29, 2016 - 14:45

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...

Choline Acetyltransferase (ChAT) – a useful Immunohistochemical marker for morphological studies of neurons

Tuesday, April 26, 2016 - 07:45

Choline Acetyltransferase (ChAT) is the enzyme that is responsible for biosynthesis of the neurotransmitter acetylcholine. The majority of acetylcholine is synthesized locally at nerve terminals where ChAT catalyzes the transfer of an acetyl group from acetyl coenzyme A to choline, a process that takes place in a single step. ChAT is expressed by cholinergic neurons in the central nervous system (CNS) and peripheral nervous systems (PNS) (4), which are distributed in many different regions of the brain, spinal chord, and retina and participate in learning, memory, movement, and vision (2,4).

ChAT antibody staining is frequently used in morphological studies of cholinergic cell populations. In the mammalian retina, for example, ChAT is expressed by subsets of both amacrine and ganglion cells (2). These ChAT-positive neuronal populations can be identified using...

The role of PARP-1 in the repair of single stranded break (SSB)

Friday, April 22, 2016 - 13:50

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...

Using RPE65 as a tool to investigate ocular gene therapies

Tuesday, April 19, 2016 - 13:17

While not life threatening, blindness and retinal disease are profoundly debilitating and greatly affect quality of life.  Understandably, gene therapy has been subject to controversy given it’s potential effects on the rest of our cellular processes.  However, a genetically diseased eye being an isolated organ quickly becomes a promising prospect for such therapies.  Specifically, RPE antibodies are powerful diagnostic tools to test the viability of these clinical treatments. 

The RPE65 protein is organized into a thin layer of cells known as the retinal pigment epithelium (RPE). This RPE provides support to the retina, which sits behind the eye and has sensitivity to light. A mutation in RPE65 leads to retinal degeneration and LCA (Leber congenital amaurosis - an autosomal recessive childhood blindness), which results in a lack of 11-cis retinal productions and an inability to efficiently form the visual pigments rhodopsin and cone opsin....

HIF-2 alpha: HIF1A's Homologue with Similar and Divergent Functions

Thursday, April 14, 2016 - 13:19

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

IRE1 alpha dependent apoptotic-signaling pathway

Monday, April 11, 2016 - 12:47

Despite in depth characterization of the role of IRE1 alpha (inositol-requiring enzyme 1 alpha) in activating the unfolded protein response (UPR) in the ER - little is known about the molecular mechanisms by which this ER protein has shown to regulate intracellular calcium levels and subsequent apoptosis. Intracellular calcium homeostasis is fundamental to many physiological processes, and an increase in Ca2+ is associated with both the early and late stages of apoptosis. A Nature article fleshed out a pro-apoptotic IRE1alphaTRAF2JNK pathway that has potential to be activated by prolonged ER stress. Specifically, prolonged IRE1-mediated activation may promote apoptosis by degrading the mRNAs that encode essential cell-survival proteins. Additional research studies have proposed similar pathways that bring in other pro apoptotic...

Caspase-3- A marker of programmed cell death

Thursday, April 7, 2016 - 13:44

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 of cell death induction. For example, caspase-3 antibodies are widely...

The subunit RelA(p65) mediates NF-kB signal transduction in multiple ways

Thursday, March 31, 2016 - 11:00

RelA (also known as p65) is an NF-kB family member and a subunit of the NF-kB transcription factor complex.  The mammalian NF-kB family has five members (NF-kB1, NF-kB2, RelA (p65), RelB, and c-Rel), each of which contains an N-terminal Rel homology domain. Active NF-kB protein complexes are dimeric (hetero- or homo-), and are made up of two family members. NF-kB signaling is activated in response to many different types of stimuli and modulates transcription of numerous downstream targets. NF-kB-mediated signaling plays known roles in inflammatory and immune responses (Hayden and Ghosh, 2008), as well as neuronal development and synaptogenesis (Boersma et al., 2011; Gutierrez et al., 2005).  Aberrant NF-kB activity has also been linked to various human diseases including a number of cancers (Perkins, 2012).

In the central nervous system, NF-kB activity is dependent on translocation to the nucleus, where the complex binds to DNA...

Three things everyone studying autophagy should know

Friday, March 25, 2016 - 09:36

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 than...

Multifaceted Roles of Matrix Metalloproteinase-2 (MMP2) in Normal and Disease State

Monday, March 21, 2016 - 13:25

MMP2 is a 72 kDa enzymatic protein and it belongs to matrix metalloproteinases (MMPs), a heterogenous family of zinc/calcium-dependent TIMPs (tissue inhibitors of matrix metalloproteinases) regulated matrix-degrading endopeptidases which are classified into collagenases (MMP-1, -8, -13, -18), gelatinases (MMP-2, -9), stromelysins (MMP-3, -7, -10, -11), elastase (MMP-12), and membrane-type matrix metalloproteinases (MT-MMP-1 through -5) (1). MMP2 involves in extracellular matrix metabolism and cleaves type IV collagen along with degrading the already denatured collagens. TIMP2 and MT-MMP1 (membrane type MMP, MMP14) regulate the activity of MMP2, and the cleavage of pro-MMP2 to active-MMP2 is dependent upon MMP14. MMP2 is expressed ubiquitously and it implicates in a diverse array of functions such as vasculature remodeling, angiogenesis, tissue repair, tumor invasion, inflammation, and atherosclerotic plaque rupture. Mutations in MMP2 gene has been linked to...

Glial Fibrillary Acidic Protein (GFAP), The Most Popular Astrocyte Marker

Wednesday, March 16, 2016 - 13:43

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].

...

Caspase 3 - a Reliable Marker for Index of Apoptosis Induction

Friday, March 11, 2016 - 15:16

Caspase-3 is one of the most important players in apoptosis signaling. It is synthesized as an inactive 32 kDa pro-enzyme and upon direct activation by Caspase-8, -9 or -10, it gets processed into its active forms, the p17-20 and p10-12 subunits. The latter are responsible for the cleavage of PARP (poly ADP-ribose polymerase), actin and SREBP, which are associated with apoptosis [1]. Because of its role in coordinating the destruction of cellular structures, Caspase 3 is often regarded as an executioner or effector caspase and considered as a marker in various immunoassays for apoptosis related experiments.

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Caspase 3 antibody

 

Tips on choosing an ideal loading control antibody for Western Blotting

Monday, March 7, 2016 - 12:56

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...

FSH R - a hormone receptor critical for both female and male reproductive systems

Monday, February 29, 2016 - 15:33

FSH R, or follicle-stimulating hormone receptor, is a transmembrane G-protein coupled receptor that is expressed in the ovaries, uterus, and testes. The ligand for this receptor, considered to be the central hormone of mammalian reproduction, is called follicle stimulating hormone (FSH) [1]. In females, FSH R is essential for proper ovarian development and follicle maturation. In males, it is required for normal spermatogenesis. Linkage analysis revealed that in females, a mutation in the extracellular ligand binding domain of FSH R segregated with a disease called hypergonadotrophic ovarian dysgenesis [2]. In males, with a mutation in the intracytoplasmatic loop that leads to ligand-independent constitutive FSH R activation, spermatogenesis is sustained in the absence of gonadotropins [3]. Based on these examples, it is...

Include tissue controls in every IHC experiment to instill confidence in your results

Wednesday, February 17, 2016 - 12:15

Interpretation of immunohistochemistry (IHC) data is difficult in the absence of appropriate controls.

To confirm staining specificity and instill confidence in your results, a positive and negative tissue control should be routinely included in IHC experiments.

Positive Tissue Control Negative Tissue Control

Control slides

 

Need help identifying a positive and negative control for your IHC experiment...

Tyrosine hydroxylase - a marker for dopaminergic neurons in the central nervous system

Monday, February 15, 2016 - 15:48

Tyrosine hydroxylase is a member of the aromatic amino acid hydroxylase (AAAH) family.  It is expressed throughout the central nervous system (CNS) and catalyzes the conversion of tyrosine to L-3,4-dihydroxyphenylalanine (L-DOPA), which can be, through a series of downstream enzymatic reactions, processed into the neurotransmitter and signaling molecule dopamine. Dopamine can then be further altered to produce norepinephrine or epinephrine. Tyrosine hydroxylase is the rate limiting enzyme in this pathway, also referred to as the catecholamine synthesis pathway. 

Antibodies that detect tyrosine hydroxylase are often used to identify dopaminergic neurons in the CNS.  In the mammalian retina, for instance, a subset of dopaminergic amacrine cells that form a single synaptic strata in the inner retina specifically express tyrosine hydroxylase and are often identified through tyrosine hydroxylase antibody staining (Wulle and...

Alpha-actin/ACTA1 - A skeletal muscle isoform mutated in various myopathies

Monday, February 15, 2016 - 15:40

Actin is an abundant cytoskeletal protein involved in a variety of cellular processes such as cell motility, cell division, and muscle contraction. Actin monomers assemble into filaments and can provide a track for transport of cargo by the molecular motor myosin (1). Alternatively, interaction with myosin allows contraction between actin filaments. This contractility is essential during cell migration and cytokinesis (1). On a larger scale, organized scaffolds of actin filaments interact with myosin to provide mechanical force during muscle contraction (1).

Actin antibodies are widely used as loading controls when measuring protein levels by performing western blots (3). While actin often serves as a normalization control in westerns or qPCR, actin antibodies also serve as important research tools for assessing the cytoskeletal network of cells or muscle fibers. For these purposes...

Tubulin alpha 4A - A ubiquitous tubulin isoform linked to ALS and infertility

Monday, February 15, 2016 - 15:23

Microtubules are a main component of the cytoskeleton and play essential roles in a variety of cellular processes. These highly dynamic tubular structures are assembled from alpha- and beta-tubulin dimers to form a complex structural network of microtubules throughout the cytoplasm. This network provides a substrate for intracellular trafficking of vesicles, organelles, and other cargo and can also facilitate cell migration. Additionally, during cell division, microtubules make up the mitotic spindle and provide the mechanical force for chromosome segregation. The diversity of microtubule functions is made possible by the large number of binding partners as well as regulation through post-translational modification. Microtubule associated proteins (MAPs), such as kinesin or dynein, can bind to tubulin and transport cargo while others alter microtubule stability. The tubulin family of proteins consists of many isoforms, each with specialized roles within the cell....

ATG11 - An important scaffolding protein in autophagosome formation and fusion

Friday, February 12, 2016 - 14:44

Autophagy is a cellular mechanism used to regulate cell metabolism and to recycle or degrade damaged organelles and proteins. This is accomplished through the engulfment of cargo in a double-membrane structure called the autophagosome. The autophagosome fuses with the vacuole or the lysosome where hydrolytic enzymes facilitate the degradation of biomolecules. Each step of autophagy is a tightly regulated process from the recognition of cargo to the assembly of the autophagosome. ATG11 is an important scaffolding protein that seems to be involved in selective autophagy. Specifically, ATG11 facilitates the fusion of autophagosomal vesicles and the vacuole by linking selective autophagy receptors with a multiprotein complex consisting of ATG1-ATG13 (1). The ATG1-ATG13 complex is inactivated under normal conditions. Starvation triggers the binding of ATG17-ATG31-ATG29 to assemble a large pentameric complex capable of facilitating autophagosome...

Three tips to successfully conjugate your own antibody

Wednesday, February 10, 2016 - 10:24

Conjugated antibodies are essential research tools for countless cell and molecular assays. While much of the time a researcher’s needs can be met with the wide variety of commercially available antibody conjugates, some applications may call for a unique reagent. For this reason some scientists opt for the flexibility of conjugating their own antibodies. While this approach may seem daunting, the availability of antibody labeling kits like the Novus Lightning-Link Kit have simplified the conjugation process and reduced the time needed to generate your own high quality antibody reagents. The Lightning-Link reagent kit allows you to efficiently label your antibody in just 30 seconds. Whether you are simply trying to save your lab a bit of money or synthesizing a unique reagent with no commercial alternatives, take into account the...

Beta-Actin's Role in Neuronal Plasticity

Thursday, February 4, 2016 - 14:29

Beta-Actin is a highly conserved protein involved in cell growth, cytoskeletal and extracellular support structures and cell migration. Because beta-Actin is ubiquitously expressed in all eukaryotic cells, it is frequently used as a loading control for assays involving protein detection, such as Western blots.

Interestingly, Beta-Actin has been shown to be associated with growth cones in developing neurite cells. While beta-Actin is one of several actins involved in the guidance of growth cones towards synaptic targets, beta-Actin mRNA has also been shown to regulate protein synthesis by binding to Vg1RBP, ultimately resulting in asymmetrical translation of beta-Actin near the site of growth cone turning.1 Beta-actin has also been shown to localize in areas associated with neuronal growth and remodeling in both developing and mature neuronal cell types. Beta-actin is primarily found in the growth cones, filopodia, cell bodies and...

Diabetes Infographic

Wednesday, January 27, 2016 - 12:54

Diabetes is a disease that impacts over 29 million people in the US (1). It is metabolic disorder that is the result of a high level or sugar or glucose in the blood. The three types of diabetes are Type 1, Type 2 and gestational diabetes, with Type 2 being the most common worldwide (2). The factors that contribute to a person getting Type 1 or Type 2 are largely unknown, but genetics, obesity and exercise have been linked to diabetes (3). Learn more about diabetes in our infographic below.

Diabetes infographic

Novus Biologicals offers 3,700+ reagents for the research of diabetes.

Resources:

  1. CDC/ 
  2. Medical News Today
  3. ...

ChREBP, a glucose sensitive transcription factor with role in glucose-lipids homeostasis and cancer

Monday, January 25, 2016 - 14:27

ChREBP (carbohydrate response element-binding protein) is a glucose responsive basic helix-loop-helix/leucine zipper (bHLH/LZ) transcription factor that binds MLX and then carbohydrate response element /ChoRE for the induction of genes involved in glycolysis, de novo lipogenesis (DNL), and fatty acid desaturation. ChREBP’s target genes includes glucokinase (GCK), fatty acid synthase (FAS), acetyl-CoA carboxylase (ACC), pyruvate kinase/liver type pyruvate kinase (PK1/ PKLR), delta-9-desaturase (SCD/SCD1) etc. ChREBP is expressed in a number of mammalian tissues such as liver, skeletal muscle, white/brown adipose, heart, kidney, cerebellum and intestine. Increased hepatic ChREBP expression has been suggested to result in development of non-alcoholic fatty liver disease (NAFLD) and obesity through the conversion of carbohydrates into triglycerides. Recent findings have suggested that besides glucose-lipids homeostasis, it implicates in pathways linked to...

SLC31A1/CTR1 - a copper transporter with important implications for platinum-based chemotherapy

Friday, January 15, 2016 - 13:04

Copper is an essential micronutrient that serves as a cofactor in numerous biological processes, but can be toxic when present in excess. Because of this, cells must tightly maintain copper levels. This includes balance between import and export of cellular copper. The major copper importer in humans is the high-affinity copper transporter SLC31A1 or CTR1. The localization of CTR1 varies between cell types, but is commonly found at the plasma membrane and intracellular vesicles [1-3].   

In addition to copper, CTR1 has the ability to transport platinum-containing drugs. One example is a chemotherapeutic drug called cisplatin that is used to treat a variety of cancers. Unfortunately, many types of cancer are resistant to platinum drugs, presenting a major obstacle for achieving maximum drug efficacy. In yeast, western blotting using a CTR1...

Dinosaur Protein Names: Infographic

Wednesday, January 13, 2016 - 11:49

Trex1 the protein is involved in DNA damage response. Tyrannosaurus rex (T. rex) the dinosaur lived during the Cretaceous Period. Raptor the protein is a regulator of mTOR activity. Velociraptors the dinosaurs lived during the Cretaceous Period. Learn more about Trex1 and Raptor proteins as well as some fun facts about the tyrannosaurus rex and the velociraptor in our infographic below.

Dinosaur protein Infogrphic

Novus Biologicals offers reagents for these proteins for your research needs:

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