Antibody News

4EBP1 and skeletal muscle protein synthesis

Wednesday, February 22, 2017 - 15:58

Eukaryotic translation initiation factor 4E binding protein 1, or 4EBP1, is an mRNA translational repressor protein that negatively regulates eukaryotic translation initiation factor 4E, or EIF4E.  EIF4E is a protein that forms a complex necessary to block the 5’ ends of mRNA with a 7-methyl-guanosine five-prime cap structure, which is important for normal translation of mRNA.  Specifically, the EIF4E complex recruits 40s ribosome subunits to scan mRNA in order to regulate protein synthesis.  When EIF4E is bound to 4EBP1, it is held in an inactive state, however phosphorylation of 4EBP1 will cause its release.  While 4EBP1 has broad implications in translational research, the following articles hone in on how a 4EBP1 antibody plays a role in the regulation of skeletal muscle protein synthesis.

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The use of Beta Actin (AC-15) as a loading control across multiple species

Friday, February 17, 2017 - 14:46

Actin is a fundamental component of the cytoskeleton, where it has the ability to create and break down actin filament formation in response to various cell needs.  Actin has six highly conserved isoforms, however beta and gamma actin are the two isoforms that are highly and ubiquitously expressed in the cell.  For this reason, measuring beta-actin levels has served as a useful control in research experiments in order to have a baseline of protein expression to compare cell manipulations to.  However, beta actin has other implications in scientific research aside from acting as a housekeeping gene, and plays a role in cell motility, development, wound healing, cell division and more.  The following articles used the Beta Actin (AC-15) antibody as a loading control in chicken, human and rat samples in order to illustrate its conservation and reliability when used across species. 

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The role of HIF-2 alpha in the progression and therapy of clear cell renal cell carcinoma

Monday, February 13, 2017 - 14:51

HIF-2 alpha, also known as hypoxia-inducible factor 2, endothelial PAS domain protein-1, and member of PAS superfamily 2 is part of the HIF family of proteins.  The HIF family is composed of HIF-1, HIF-2 and HIF-3, where HIF-2 is a dimeric protein that consists of an alpha and beta subunit.HIF-2 alphais activated in the presence of oxygen deprivation, or hypoxia, via prolyl hydroxylase-domain enzymes (PHDs) that facilitate its degradation.  HIF-2 alphais expressed in endothelial and intestinal cells, as well as in tumor tissue of a variety of prominent cancers, and is not typically detected in normoxic conditions.  Given the fact that loss of HIF-1 alpha was a notable hallmark of renal cell carcinoma, HIF-2 alpha and the hypoxia mediated gene therapy has been an area of interest in...

Dual applications of a c-Myc antibody in mitochondrial research

Thursday, February 9, 2017 - 08:10

c-Myc, a proto-oncogene, has documented involvement in cellular differentiation, cell growth, cell death and tumor formation.  Target genes of the Myc family include those that participate in cell survival, translation, transcription, metabolism and more.  On a more specific level, c-Myc is a transcription factor that can both activate and repress its target protein by way of DNA modifications.  This allows for the use of a c-Myc antibody in two manners; it can be used to monitor the actual c-Myc protein expression levels, or, it can be used as a probe against a c-Myc protein tagged to another protein of interest. These interactions can be exploited when trying to detect proteins that c-myc interacts with if antibodies against the protein are not available. The structure of c-Myc is what allows for its duality, where its basic-helix-loop-helix-leucine zipper c-term domain can mediate DNA specific binding.  The...

CiteAb 2017 Antibody Awards: The Lifetime Achievement Award Recipient Karen Padgett

Monday, January 30, 2017 - 12:53

The CiteAb Lifetime Achievement Award recognizes and promotes an individual who has made an outstanding contribution to the research antibody industry over a sustained period of time.

We're delighted to announce that Karen Padgett, founder of Novus Biologicals, has been recognized for her lifetime achievement in the research antibody industry. Karen is currently the VP of the Antibody Business Unit (representing both Novus and R&D Systems) and Digital Marketing at Bio-Techne. Karen has earned her place as a global leader in the research antibody industry, through her unfailing commitment, drive and positivity over the last two decades, and without ever compromising the integrity that defines her company as an industry leader.

Citeab award

Over 20 years of...

IHC vs ICC vs IF – Do you know the difference?

Monday, January 30, 2017 - 08:22

Immunohistochemistry (IHC), Immunocytochemistry (ICC) and Immunofluorescence (IF) all utilize antibodies to provide visual details about protein abundance, distribution and localization. These terms are often confusing and are sometimes mistakenly used interchangeably.  Thus, it is important to understand the fundamental differences between these various techniques. 


IHC-Fr (on left): Cadherin‑17 was detected in perfusion fixed frozen sections of mouse intestine using rabbit anti-mouse Cadherin‑17 monoclonal antibody (Catalog # MAB8524...

The importance of using controls in immunohistochemistry

Monday, January 23, 2017 - 08:52

Immunohistochemistry (IHC) is a widely applied experimental method used to examine tissue antigen expression and behavior with the use of an antibody conjugated to a secondary tag for visualization.  IHC consists of a tissue preparation phase, an antibody-staining phase, and a result analysis phase - all of which may lead to skewed results if not properly performed.  One way to ensure that IHC staining results are in fact demonstrating protein behavior and not a side effect the experimental process is to use IHC controls within each experiment.  By doing so, you can feel confident in your protein integrity, experimental methods and primary and secondary antibody efficacy. 

To begin, the manner in which tissue is harvested, dissected and mounted to a microscope slide is very important.  It is vital that the tissue is collected rapidly in order for the proteins to...

The role of p53 in UV radiation DNA damage and subsequent tumorogenesis

Friday, January 20, 2017 - 13:19

p53, the protein product of the tp53 gene, is one of the most widely studied tumor suppressor proteins in cancer research.  p53 is unique in that it demonstrates both tumor suppressive and tumor progressive properties depending on whether it is functional or mutated.  The most common mutation in the p53 protein that leads to lack of tumor suppression activity is a missense mutation, however frameshift or nonsense mutations are also common.  In fact, mutant p53 has exhibited dominant negative inhibition of the wild type version of the protein, demonstrating the fact that the p53 pathway has both positive and negative feedback loops.  The p53 pathway is a large signal transduction pathway that affects DNA replication and division, apoptosis, cellular senescence and is composed of at least seven negative feedback loops and three positive feedback loops.  Although p53's major influence is in cancer, the following articles describe how a...

The application of CD31/Pecam-1 (MEC 7.46) in breast cancer research

Monday, January 16, 2017 - 15:27

CD31/PECAM-1, or platelet endothelial cell adhesion molecule 1, is a 130-kDa glycoprotein expressed on vascular and hematopoietic cells.  Depending on the cell type, CD31/PECAM-1 expression can be largely localized to cell junctions, playing a role in cell adhesion.  Aside from its role in cellular adhesion, CD31/PECAM-1 is also a large player in a variety of signaling pathways, such as angiogenesis, cell migration, leukocyte transmigration and more.  Specifically, the association and indication of angiogenesis in breast cancer is of interest, given that extensive research shows angiogenesis is involved in breast cancer development, invasion and metastasis.  The use of a CD31/PECAM-1 primary antibody in experimental studies to quantify tumor neovascularization is a popular approach to measure microvessel density.

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The use of actin as a loading control in research on fruiting-body development and vegetative growth in Sordaria macrospora research

Monday, January 9, 2017 - 14:46

Sordaria macrospora is a filamentous fungus that serves as very useful system for scientific research due to a short life cycle and easy manipulation.  Just like any other model organism, it is important to have an effective loading control to validate experiments in the Sordaria macrospora.  In addition, the growth and morphogenesis of filamentous fungi is dependent on actin organization.  Actin is a very abundant protein across biological species and can transition between monomeric and filamentous states.  Actin has an alpha and beta isoform, both of which share nearly 98% percent sequence homology with total actin.  Actin recruits a variety of actin regulators in order to carry out many cellular functions, including cell division, cell motility, cell growth and differentiation, muscle contraction and more.  The following articles use an actin antibody as a loading control in the study of...

The C99 fragment of amyloid precursor protein (APP)

Friday, January 6, 2017 - 08:11

Alzheimer’s Disease (AD) is a neurodegenerative disorder that is characterized by an abundance of the beta-amyloid peptide in the brain.  When AD was first discovered, it was determined that beta-amyloid was produced as a result of the proteolysis of the amyloid precursor protein (APP).  Aside from its role in AD, the single-pass transmembrane APP has a high expression level in the brain and tends to concentrate at the synapses of neurons.  Because of this localization, it has been suggested that APP plays a role in synapse formation and potentially plasticity.  However, the exact function of APP is not known.  While there is currently no answer as to why APP accumulates in the brain of elderly individuals, current therapeutic approaches tend to focus on treatments that affect APP processing.  Over the past few years, the structure of APP has been closely studied.  From this research came the discovery that amyloid-beta production requires two cleavage steps...

Pyruvate Dehydrogenase E1-alpha subunit: Understanding the Catalysts in Glycolysis

Thursday, December 29, 2016 - 14:01

Pyruvate Dehydrogenase E1-alpha subunit (PDHA1) is one of multiple enzymes in a mitochondrial complex, called the Pyruvate Dehydrogenase (PDH) complex, involved in the transition between glycolysis and the tricarboxylic acid (TCA) cycle during cellular respiration. The PDHA1 enzyme catalyzes the reaction that produces acetyl-CoA and CO2 from pyruvate (1). PDHA1 also regulates the PDH complex through reversible phosphorylation.

PDHA1 antibody

Pyruvate Dehydrogenase E1-alpha subunit [p Ser293] Antibody [NB110-93479] - Pyruvate Dehydrogenase E1-alpha subunit [p Ser293] antibody (1:250) was tested in HeLa cells with Dylight 488 (green). Nuclei and alpha-tubulin were counterstained with DAPI (blue) and Dylight 550 (red)....

The dynamic use of a PCNA antibody in fish, porcine and primate species

Tuesday, December 27, 2016 - 14:39

Proliferating cell nuclear antigen (PCNA) plays a crucial role in nucleic acid metabolism as it pertains to DNA replication and repair.  Most noted for its activation of subunits of DNA polymerase, it has also been found to interact with cell-cycle progression proteins.  Modifications of PCNA as a result of cellular response put PCNA in a pivotal position with DNA replication, DNA damage, and chromatin structure and function.  In response to DNA damage, PCNA is ubiquitinated and becomes part of the RAD-6 dependent DNA repair pathway, where it acts as a substrate with a variety of proteins. In DNA replication, PCNA acts as a sliding clamp that localizes proteins to DNA strands.  Additionally, the presence of PCNA at DNA maintenance sites where delayed replication forks are found further points to its role in DNA damage and repair. The PCNA mouse monoclonal antibody (PC10) from Novus Biologicals has been effective to...

The use of a GFP antibody for research applications in transgenic C. elegans, GFP tagged yeast and porcine model

Monday, December 19, 2016 - 15:32

GFP, or green fluorescent protein, is a chemiluminescent protein derived from Aequorea jellyfish that was first discovered by Osamu Shimomura.  It was soon after established that the emission spectra of GFP was right around 509nm, or the ultraviolet color range.  The GFP gene is often used to form expression constructs in order to closely follow protein behavior, cellular differentiation, protein localization and more.  The following articles employed a GFP antibody in conjunction with various other GFP construct techniques to strengthen their research findings. 

GFP antibody

Immunocytochemistry/Immunofluorescence: GFP Antibody [NB600-308] - Analysis of GFP in transgenic mouse pancreas in OCT. Verified customer review from...

The Key Benefits of Indirect Detection

Tuesday, December 13, 2016 - 11:50

Even though the direct detection method is becoming more popular for immunofluorescence (IF) and flow cytometry experiments, the indirect detection method still remains the preferred choice for many other applications. In direct detection, the labeled primary antibody is responsible for both binding and detection of the antigen of interest. In indirect detection, this process is broken down into at least two distinct steps – (i) an unconjugated primary antibody forms a complex with the antigen, (ii) a labeled secondary antibody, interacting with the constant region of the primary antibody, facilitates detection.

HAl-1 antibody

HAI-1 was detected in paraffin-embedded sections of human lung cancer using goat anti-human HAI-1 ectodomain antigen affinity-purified polyclonal antibody (Catalog # ...

Top 4 reasons: why use CRISPR-Cas9 antibodies and how?

Friday, December 9, 2016 - 14:37
1. Verification of the success of transfection

Why- If the CRISPR-Cas9 transfection is not successful, it would not be relevant to relate the observations from transfected cells to the expected outcome of gene editing experiment.

How- CRISPR-Cas9’s successful transfection can be verified through its detection at the protein level by employing Western blot or confocal staining analysis of cells that were subjected to CRISPR-Cas9 transfection.

2. Localization of CRISPR-Cas9 at subcellular level

Why- CRISPR-Cas9 must translocate to the nuclei of the transfected cells for executing its nuclease activity on the genomic DNA. Therefore, one must check that the Cas9 protein is actually being delivered into the nucleus.

How - The...

The effects of curcumin on IKB Alpha and the NFkB signaling pathway

Wednesday, December 7, 2016 - 14:15

The IKK complex, or inhibitor of NFkB kinase, is composed of IKK alpha and IKK beta.  These kinases are at the core of the NFkB signaling cascade.  The NFkB family is made up of transcription factors that are kept inactive in the cytoplasm through inhibitory IkB proteins. When various stimuli are activated, these IKK proteins go through phosphorylation, leading to polyubiquitination and subsequent degradation.  Without the presence of these inhibitory factors, NFkB can now enter the nucleus and turn on the transcription of a variety of genes that influence cellular proliferation, immune response, apoptosis, cancer pathways and more.  Recently, research has shed light on the yellow pigment from Curcuma longa, curcumin, as having potential therapeutic properties through its antioxidant and anti-inflammatory characteristics.  The following experiments use an IKB alpha antibody to investigate the effects of...

Transportin 1 and heterogeneous nuclear ribonucleoprotein D (hnRNPD)

Tuesday, November 29, 2016 - 14:49

Transportin 1, also known as Karyopherin- β 2 or Importin- β 2, is part of the β-karyopherins family, which consists of importins and exportins responsible for the active transport of proteins between the nucleus and cytoplasm.  Transportin 1 is composed of twenty HEAT (or a tandem repeat protein structural motif comprised of two alpha helices that end with a short loop) stacks that form a helix.  In the presence of Ran-GTP, Transportin 1 undergoes a conformational change to release the cargo it’s transporting.  Transportin 1 is known to bind and transport hnRNP A1, or heterogeneous ribonucleoprotein A1.  hnRNPD, the ribonucleoprotein highlighted in the following articles, is a protein that binds RNA molecules with AU-rich elements (AREs).  It can also function as a transcription factor when bound to DNA sequences, and has roles in mRNA biogenesis and metabolism.  The following research...

Novus Food Drive

Wednesday, November 23, 2016 - 11:22

This year the Novus team participated in an office food drive to help feed the homeless and families in need.  

In the Colorado, 1 in 8 people may not have enough money to buy food. For the children in Colorado, 1 in 5 may not know where their next meal may come from (source Hunger Free Colorado). 

We were able to collect over 247 pounds of food including:

  • 3 large turkeys
  • 27 pounds of potatoes
  • 30+ canned food items from veggies to cranberry sauce
  • 12+ boxes of stuffing

All the donations were delivered to Aurora Interchurch.

Thanks to all who donated and we hope that everyone has a Happy Thanksgiving!

Novus office food drive

Happy Thanksgiving!

AMPK Alpha 1 and lipid metabolism of adipocytes

Tuesday, November 22, 2016 - 13:32

AMP-activated protein kinase (AMPK) is best known as a sensor of oxidative stress.  AMPK is activated by increased intracellular AMP levels, which are a result of alterations in cellular metabolism from causes such as hypoxia, changes in ATP, senescence and more.  In cell stress models, AMPK can protect cells from reduced ATP production by altering ATP biosynthetic pathways.  Furthermore, AMPK has implications in reducing inflammatory reactions in apoptosis pathways. AMPK is also activated during exercise and other situations where lipolysis is enhanced.  While this mechanism is not fully understood, research has shed light on the phosphorylation of the Thr-172 residue of the AMPK Alpha 1 subunit by upstream kinases.  An AMPK Alpha 1 antibody has been used to further investigate how adipose tissue and energy regulation play into AMPK behavior and lipid metabolism.  The following articles discuss the use of the...

Required proteins for p62/SQSTM1 regulation and a role for p62/SQSTM1 in neuronal autophagy

Thursday, November 17, 2016 - 09:44

Autophagy is a crucial cellular process that clears the cell of protein aggregates, toxins, and damaged cell products. Accumulation of toxins, damaged cell products and unwanted proteins has been proven to play a role in aging and many forms of disease and cancer. p62/SQSTM1, or sequestosome-1, is an autophagosome receptor that interacts with cargo tagged for degradation in order to turn on selective autophagy.  p62/SQSTM1 binds LC3 through the LC3 interacting region (LIR), which is necessary for degradation of sequestosomes.  p62/SQSTM1 is also required for the formation and degradation of polyubiquitin-containing bodies in autophagy. Outside of p62/SQSTM1’s role in autophagy, it has also been implicated in cell signaling that impacts differentiation, apoptosis and immune response.  Here we take a closer look at the proteins in the brain that may be required for autophagy, as well as defective autophagy in neurons, with the use of a...

The role of LC3B and autophagy in alcohol induced liver disease

Friday, November 11, 2016 - 14:05

Autophagy is a crucial intracellular pathway that manages the degradation and recycling of long-lived proteins in the cell. The LC3 (or light chain 3) family is composed of three members, LC3A, LC3B and LC3C. Upon autophagy induction, LC3 is cleaved, causing the release of a C-terminal glycine that is required for phospholipid conjugation.  This process is vital to the formation of the autophagosome, a double membrane structure that delivers proteins to the lysosome during autophagy. This process has made LC3 a strong indicator of autophagy; therefore, the use of LC3 antibodies in autophagy research is extremely efficient.  LC3 and autophagy are important parts of development, differentiation, survival and homeostasis.  A link between autophagy and hepatology has been previously established and found to have anywhere from an anti-tumor effect to a pro-survival activity during liver ischemia characterized by nutrient...

Five key tips for a better multicolor immunofluorescence staining

Tuesday, November 8, 2016 - 08:56
  1. Multicolor immunofluorescence staining is best carried out by sequentially incubating cells with unlabeled-primary and labeled-secondary antibodies. However when options are limited, it may also be performed by simultaneous incubation of cells with directly labelled primary antibodies.  
  2. When picking fluorochrome colors for labeled primary or secondary antibodies, one must consider their fluorescent emission spectrum. Ensure that there is a minimal to negligible emission-spectra overlap between the selected fluorochromes. This will help eliminate bleed through effects during staining analysis.  

  3. It is recommended to use the dimmest fluorochrome from one's available options to label the most abundant protein. Conversely, one should try detecting the least abundant protein with the fluorochrome having the highest quantum yield. These tips will help to efficiently/accurately detect multiple proteins...

Further unraveling the role of gamma H2AX in DNA damage response

Friday, November 4, 2016 - 13:13

Our genome experiences a moderate amount of DNA damage in our cells on a daily basis.  This DNA damage can be in response to external environmental factors, or be a result of our internal metabolic processes going awry.  While normal rates of DNA damage are not an immense threat to our cell processes, DNA damage in critical genes can lead to a variety of disease, including cancer and tumor formation.   After induction of DNA damage (for example, in the form of double strand breaks), phosphorylation and recruitment of the H2AX protein occurs.  This phosphorylation produces gamma H2AX, which is crucial for activating the DNA damage response (which in turn assembles DNA repair proteins at the mutation site) and creating checkpoint proteins to arrest the cell cycle.  Gamma H2AX antibodies have been utilized to measure the toxic effect of a treatment or drug at the genotypic level, and are widely applied to a variety of...

The role of HIF-1 Alpha signaling in the retina under hypoxic conditions

Thursday, November 3, 2016 - 13:52

Hypoxia inducible factor 1 (HIF-1) is a protein that plays an essential role in hypoxia, or low levels of cellular oxygen. HIF-1 is a heterodimeric protein that consists of a constitutively expressed beta subunit and oxygen related alpha subunit.  Both subunits have a basic helix-loop-helix domain that leads to dimerization, where HIF-1 alpha carries an oxygen-dependent degradation (ODD) domain.  Outside of the role of HIF-1 in hypoxia adaptation, it is also involved in cancer, inflammation and metabolism.  The HIF-1 alpha subunit is specifically degraded in normal oxygen environments, where it is not active and functioning until a hypoxic environment is sensed.  In some cases, HIF-1 alpha expression can be quite high, leading to angiogenesis and increased blood flow.  The link between HIF-1 alpha activity and retinal pathways has long been established, given that photoreceptors in the retina require a large amount of oxygen to function properly.  The use of a...

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