Antibody News

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...

CRISPR/Cas9: Keep your friends close, but your viruses closer

Tuesday, October 25, 2016 - 15:50

"CRISPR", or clustered regularly interspaced short palindromic repeats, is an ancient bacterial mechanism that prevents the invasion of foreign pathogens to a host organism.  Specifically, the CRISPR sequence has been identified as a single DNA sequence that is repeated with unique sequences (found to be that of viruses) in-between.  Thus, bacteria have created an environment that allows them to recognize and attack viruses in case of a re-invasion.  "Cas", or CRISPR-associate protein, is the second part of this defense mechanism, and is responsible for cutting targeted DNA and viruses.  Cas9 is the best-known "Cas" enzyme and originates from Steptococcus pyogenes (or strep throat).  Together, CRISPR/Cas9 work to fill the CRISPR region with foreign DNA and turn on Cas9's splicing machinery.  Next, the copied viral RNA and Cas enzymes identify viral material and slice it apart to avoid further replication.  Since the discovery of this...

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. 

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