Novus Biologicals Blog

Amyotrophic Lateral Sclerosis Infographic

July 29th, 2014

Amyotrophic lateral sclerosis is a neurological disease which impacts motor neurons that are involved in muscle movement throughout the body. The progressive degeneration of neurons causes weakened muscles and can lead to paralysis. There is no cure for ALS, but riluzole has been used to help with the damage to the motor neurons.

AMYOTROPHIC LATERAL SCLEROSIS (ALS)

 

Resources:

  1. ALSA.org 
  2. Lougehrig.com 
  3. NIH.gov 
  4. ALSA.org
  5. Mayoclinic.org 
  6. MDA.org 
  7. NIH.gov 
  8. ALShopefoundation.org 

  Download the PDF version of the ALS infographic

By: Lisa Ikariyama; Design: Kim Mesman

HIF-1 beta: A dimerization partner of HIF-1 alpha required for an adaptive response to hypoxia

July 28th, 2014

Hypoxia contributes significantly to the pathophysiology of major categories of human disease, including myocardial and cerebral ischemia, cancer, pulmonary hypertension, congenital heart disease and chronic obstructive pulmonary disease. Hypoxia-inducible factor 1 (HIF-1) is a nuclear protein involved in mammalian oxygen homeostasis. It is a heterodimer composed of HIF-1 alpha and HIF-1 beta subunits which are bHLH proteins of PAS (PER, ARNT, SIM) family. HIF-1 beta forms heterodimer with AHR, AHRR, HIF1 alpha and EPAS1/HIF2 alpha as well as with other bHLH proteins and its dimerization is required for efficient DNA binding.

HIF-1 beta is encoded by the ARNT (Aryl Hydrocarbon Receptor Nuclear Translocator) gene. It is able to partner with AhR to form a heterodimeric transcription factor complex which is required for AhR activity. This protein is required for the ligand-binding subunit to translocate from the cytosol to the nucleus after ligand binding. The complex then initiates transcription of genes involved in the activation of PAH pro-carcinogens. The heterodimer with HIF1A or EPAS1/HIF2A functions as a transcriptional regulator of the adaptive response to hypoxia.

Hypoxia-inducible factor 1 (HIF-1) is critically involved in cancer cell hypoxia adaptation, glycolysis, and angiogenesis. HIF-1 beta is associated with HIF-1 functions as a dimerization partner of HIF-1 alpha, and is on the other hand associated with carcinogenesis via dioxin signaling. A study using Novus’ HIF-1 beta antibody (NB100-124) suggested that HIF-1alpha and beta share common signaling pathways for nuclear protein accumulation. Regulation of HIF-1beta protein expression was investigated in human prostate cancer (PCA) cells. (1)

Western Blot: HIF-1 beta Antibody

Western Blot: HIF-1 beta Antibody

The Ah receptor nuclear translocator protein (ARNT) is required for Ah receptor function. ARNT is shown to be a structural component of the XRE binding form of the Ah receptor. Furthermore, ARNT and the ligand-binding subunit of the receptor were extracted as a complex from the nuclei of cells treated with ligand. Arnt contains a basic helix-loop-helix motif, which may be responsible for interacting with both the XRE and the ligand-binding subunit. (2)

Novus Biologicals offers HIF-1 beta reagents for your research needs including:

PMIDs:

  1. 11394885
  2. 1317062

Essential to Death: ATG5 (autophagy protein 5, apoptosis-specific protein ASP)

July 22nd, 2014

The ATG5 protein belongs to the ATG autophagy regulator family. This family controls the highly conserved cell’s homeostatic response to a wide variety of both self- and foreign-originating cellular stimuli. ATG5 itself is ubiquitously expressed in most cells and most often found co-localized with the cytoplasmic non-muscle actin protein under normal resting conditions. Upon activation of apoptosis, ATG5 expression is then dramatically intensified, with ATG5 directly complexing with its ATG family members to produce autophagosomes. An intriguing application of the ATG5 antibody was done by Herd’s group where they used it to characterize the effect of silica nanomaterials on both cell uptake and toxicity in epithelial phagocytic cells1. The antibody allowed this group to monitor the effect of physicochemical manipulation of these silica-based nanoparticles on lysosomal formation and function, autophagic like activity, and compartmental fusion and recycling.

Western Blot: ATG5 Antibody

Western Blot: ATG5 Antibody

A protective role for autophagy in chronic obstructive pulmonary disease (COPD) in response to tobacco smoke was identified by for Fujii et al who performed ATG5 antibody immunoblotting as an autophagy marker2. Kang’s group used the ATG5 antibody in their studies linking autophagy to interleukin 2 (IL2)-induced cell growth, apoptosis, and immmunoregulation – a novel link for autophagy in survival regulation and wound healing3. A recent Nature Communication publication highlights the use of the ATG5 antibody to demonstrate an unconventional function of ATG proteins such as ATG5 and ATG7 as molecular scaffolds in the ubiquitous extracellular signal-regulated kinase (ERK) signal cascade4. Yen’s group used the ATG5 antibody in both immunoblotting and immunohistochemistry experiments examining the effects on autophagy modification with drugs such as rapamycin and activated protein C (APC) in treating and repairing sepsis-induced acute lung injury5.

Novus Biologicals offers ATG5 reagents for your research needs including:

PMIDs

  1. 21342660
  2. 22934255
  3. 23195496
  4. 24240988
  5. 23434181

BRCA1 – A Critical Tumor Suppressor Gene in Women

July 21st, 2014

Breast cancer 1, early onset (BRCA1) is a well-known tumor suppressor gene that was originally discovered due to its link with early-onset breast and ovarian cancer in women. The BRCA1 protein contains the following domains: RING finger, RAD51-interaction, and BRCT (BRCA1 C-terminus). The N-terminus RING domain enables binding to several proteins – including BARD1 (BRCA1-associated RING domain protein) – allowing the formation of heterodimers. The RING finger is important for tumor suppressor activity. The RAD51-interaction domain is involved in DNA double-stranded break (DSB) repair. The loss of RAD51 binding increases cancer risk due to increased increments of damaged DNA. The BRCT functions as a transcriptional activation domain and is involved in cell-cycle control and DNA repair. BRCA1 has been implicated in a wide range of cancer-related activities, such as cell cycle progression, DNA repair, DNA damage-responsive cell cycle checkpoints, transcription regulation, ubiquitination, chromosome remodeling, and apoptosis. BRCA1 defects within an individual’s genome increases the likelihood of occurrence of breast cancer, familial breast-ovarian cancer type 1 (BROVCA1), ovarian cancer, and pancreatic cancer type 4 (PNCA4).

Western Blot: BRCA1 Antibody

Western Blot: BRCA1 Antibody

Okada et al paired confocal microscopy and immunoblotting with BRCA1 antibody to monitor how cell cycle differences in BRCA1 phosphorylation affect its subcellular localization1. Oncologists employed the BRCA1 antibody in their ataxia telangiectasia mutated (ATM) model to closely examine DNA damage and the triggered assembly of BRCA1, BAAT1, and ATM2. Genomic research from Kluk’s group relied upon the BRCA1 antibody to help them confirm expression of BRCA1 and an isoform of the DLX4 homeobox gene (BP1) in familial versus sporadic breast cancers3. Their studies suggest that BP1 plays an important role in negatively regulating BRCA1 and further work is needed to see if BP1 is a usable drug therapy target. Marin et al from UCLA used the BRCA1 antibody in their assessments of radiosensitivity using kinetic profiles4. They believe they have identified a means of globally identifying defective DNA repair pathways in radiosensitive cells. Recent results from Canada used BRCA1 antibody immunofluorescence to detect alternation open reading frames (ORFs) translation products by subjecting a synthesized proteome database to mass spectrometry5.

Novus Biologicals offers BRCA1 reagents for your research needs including:

PMIDs

  1. 12427729
  2. 16452482
  3. 20877436
  4. 21722985
  5. 23950983

6 Fictional Character Protein Names

July 18th, 2014

Check out our list of some of the funniest fictional character protein names including GOLM1, SMURF1, HOMER1, Hunchback, Lilliputian, Pokemon, Tinman Paralog, and Draculin. What are your favorite or funniest protein names? Share them in the comments; we would love to hear any others that you come up with!

Funny protein names

Novus Biologicals offers reagents mentioned in this post including:

 Download the Fictional Character Protein Names Infographic

Resources:

  1. Genecards
  2. Uniprot
  3. Genatlas
  4. Uniprot
  5. Uniprot
  6. Wikigenes
  7. Uniprot
  8. NCBI
  9. Sciencegateway
  10. Genatlas
  11. Uniprot
  12. Genecards
  13. Uniprot
  14. NCBI
  15. USBio
  16. Unimass

By: Lisa Ikariyama; Design: Kim Mesman

CD4 – An Important Co-receptor Assisting TCRs

July 17th, 2014

The cluster of differentiation 4 (CD4) protein is a surface, type I membrane-embedded glycoprotein that is found on a wide range of cells: T-lymphocytes, B-cells, macrophages, granulocytes, and developmentally-dependent regions specific to the brain. It interacts with major histocompatibility complex (MHC) class II anti-genes, serves as the human immunodeficiency virus (HIV) receptor, and can initiate/augment early phase T-cell activation.  In immune-mediated and central nervous system (CNS) infectious diseases, CD4 indirectly mediates neuronal damage. There exists three identified CD4 splice variants, each encoding a different CD4 isoform: one with a predicted transmembrane (TM) region and two with predicted signal peptides.

Immunohistochemistry-Paraffin: CD4 Antibody

Immunohistochemistry-Paraffin: CD4 Antibody

Yasuda’s group used the CD4 antibody to demonstrate that a concerted, combined blockade of both VEGFR2 and PDL in murine adenocarcinomas was synergistic and highly anti-tumorigenic1.  The same group more recently published additional follow-up studies with the CD4 antibody demonstrating similar findings in a Herpesvirus entry mediator (HVEM) system where they examined human esophageal cancers2. Hwang et al employed the CD4 antibody to determine the role of the natural compound acidic polysaccharide of Panax ginseng (APG) in autoimmune demyelinating diseases such as encephalomyelitis and multiple sclerosis (MS)3. Immunohistochemistry with the CD4 antibody was used in studies from a Chinese laboratory, where Notch1-dependent signaling and mechanisms were mapped and characterized in abdominal aortic aneurysms (AAA)4. The CD4 antibody was also used to monitor the effects of programmed freeze-thaw cycles on cryopreserved tissues5. The findings from that study provide support for utility of such a cycling method as a means of reducing immunogenicity and therefore decreasing allograft immune rejection.

Novus Biologicals offers CD4 reagents for your research needs including:

PMIDs

1: 23600839, 2: 24249528, 3: 21524666, 4: 24358274, 5: 22127304

Cyclooxygenase 2 (COX2, PGHS2) – I stay with Inflammation!

July 14th, 2014

COX2 is an inducible dimeric enzyme belonging to the prostaglandin G/H synthase family that enables cells to respond to growth factors, tumor promoters, and cytokines. Prostaglandins are synthesized through the creation of cyclic endoperoxides from arachidonic acid and COX2 catalyzes the committed step in the biosynthetic pathway.  Initially identified as an immediate early growth response gene, COX2 is induced by a wide variety of stimuli such as lipopolysaccharide (LPS) and peptoglycan (PGN). COX2 is highly expressed in activated macrophages and plays a role in immune response regulation. It has been implicated as a mediator of inflammation as well as a modulator of prostanoid signaling in activity-dependent plasticity. It holds promise as both a marker in immune response regulation studies and an inflammatory disease therapeutic target.

Immunohistochemistry-Paraffin: COX2 Antibody

Immunohistochemistry-Paraffin: COX2 Antibody

Additionally, COX2 expression is markedly increased in 85-90% of human colorectal adenocarcinoma whereas the related protein COX1 is unchanged. Wada’s group used the COX2 antibody to study compensatory signaling mechanisms for COX2 in a variety of tissues (brain, macrophages, skin, kidney, and spleen) 1. A research team from China performed immunohistochemistry using the COX2 antibody to identify those genes induced early in oral carcinogenesis through hierarchical clustering based on microarray analysis2. Their data identified COX2 and NFkB in addition to cytokines and cytokine receptors. Immunoblotting with the COX2 antibody helped Kogiso et al understand the role of PPAR gamma in mycobacterial pulmonary inflammation3. Paiotti’s group used the COX2 antibody to evaluate COX1 and COX2 expression in ulcerative colitis (UC) disease to correlate it with clinicopathological parameters4. The COX2 antibody also allowed biochemists to measure the effect of dietary fish oil fatty acids on COX- and lipoxygenase-derived products in the colonic mucosa5.

Novus Biologicals offers COX2 reagents for your research needs including:

PMIDs

  1. 19758985
  2. 22561872
  3. 22696146
  4. 17653443
  5. 21937210

C9RANT – A Potential Target in Neurodegenerative Diseases

July 11th, 2014

C9RANT is a newly discovered protein-like chain that holds great potential promise as a benchmark for measuring disease activity and therapeutic response for patients with the devastating conditions of amyotrophic lateral sclerosis (ALS) and dementia. Mutations within a non-coding region of the C9ORF72 gene are responsible for 20-40% of cases of familial ALS1. This class of mutations generates repetitive, high molecular weight RNA chains which are insoluble and aggregate within the brain as C9RANT accumulations. As with the tau protein in Alzheimer’s disease, C9RANT is not found in either healthy people or those with other neurologic diseases.

Immunohistochemistry-Paraffin: C9RANT Antibody

Immunohistochemistry-Paraffin: C9RANT Antibody

Ash et al published their exciting results on the first identification and characterization of these protein aggregates in Neuron. They used the C9RANT antibody for multiple assays including immunohistochemistry, immunoblotting, and immunofluorescence. Further work must be done to determine if C9RANT is just a biomarker or directly involved in causing cell death and toxicity. Very recent studies in dementia from Dickson’s group at the Mayo Clinic suggest that the clinical and pathological occurrence of C9ORF72 repeats can be further widened and expanded to include other diseases such as progressive amnestic dementia and depressive pseudodementia2,3.

Novus Biologicals offers C9RANT antibodies for your research needs.

PMIDs

  1. 23415312
  2. 23922030
  3. 24756204

Your Expectations, Experience, and Evaluations of Antibody Customer Service Programs

July 10th, 2014

What do scientists think about customer service at places like Novus, companies that sell the antibodies that are often critical components of a wide variety of experiments? Novus ran a survey to find out, and here’s what we uncovered.

There is a clear disconnect between expectations of antibodies, and how they perform in real world experiments. Seventy percent of respondents said that they expected their antibodies to work every time, but only 35% reported that this is what actually happens.

When an antibody fails, 69% of respondents expected a full refund of the cost of the antibody. But less than half felt that they had been offered a refund or a replacement product that was warranted given experimental circumstances. This finding really surprised us because Novus offers a 100% guarantee on every product—so to the majority who felt that they didn’t get a refund or replacement, talk to us. We stand behind our products and want them to work for you.

The good news is that most technical support teams are doing good work—sixty-four percent of survey-takers also expected the technical support team to troubleshoot what may have gone wrong with their antibody, and this expectation was generally met.

More than 80% of respondents also agree that customer service representatives were courteous, easy to understand, and eager to assist with their antibody problems. Seventy-seven percent also said that the representatives were technically knowledgeable about their antibody products, and 63% said that the representatives provided useful experimental protocol advice.

Novus’s Customer Service Programs

We were excited to see the enthusiasm for the different customer service programs we tested, especially since we already offer the Antibody Concierge service and the Laverne tool—so go ahead and take advantage of these services. Here are the top three initiatives that were favored by at least 90% of survey-takers.

Antibody Concierge. The most popular proposed program is a free service designed to help scientists look for rare and unusual antibodies. Working off a user’s specifications, Novus will search for all known antibody manufacturers worldwide and then tell the user how their antibody can be purchased — even if Novus doesn’t offer that product. Almost every respondent (93%) considered the concierge service a valuable tool. “It will be [the] perfect service if you can help me find and order any antibody, which is a boring time waste, but [still] critical work,” one researcher noted.

Laverne Tool. Ninety-one percent of respondents said that Laverne — an easy-to-use point-and-click tool designed to help users find information about unfamiliar proteins — will be valuable to their research. Scientists can type in a gene, pathway, or disease they’d like to learn more about, and then find related information on their screen — with direct links to PubMed. One respondent cautioned, however, that Novus “needs a better way to differentiate [Laverne], because most scientists are able to use search engines to find this kind of information by themselves.”

Antibody Quality Callback. Similarly, 90% of participants thought that an antibody quality callback program will offer value by giving scientists an idea of how well certain antibodies are working for their peers. Through the callback program, Novus will email users a one-question survey on antibody performance one month after they make an antibody purchase. The results will then be posted in the Key Performance Indicators section of Novus’s website.

Novus appreciates all the valuable input from antibody users and is excited to see so much interest in its customer service initiatives. What else do you think Novus can do to improve your satisfaction around customer service? Let us know in the comments.

Aromatase – A Key Enzyme in the Biosynthesis of Estrogens

July 9th, 2014

The enzyme, aromatase, belongs to cytochrome P450 family of monooxygenases known for their key role in drug catabolism and cholesterol/steroid synthesis. Aromatase uses a heme-group as a co-factor to catalyze the formation of aromatic C18 estrogens from C19 androgens. This conversion includes that of testosterone into estradiol as well as androstenedione to estrone. Aromatase is a peripheral membrane protein. In addition to being found in the female reproductive tract and adipose tissue, it can also be found in testis, liver, and brain – as well as certain tumors including breast cancer. During ontogenesis, it plays an important role in CNS and gonad development, as well as sex differentiation. Defective aromatase leads to the aromatase excess syndrome (AEXS) and aromatase deficiency (AROD). Additionally, obesity has been linked to abnormally high aromatase expression in mammary tissues, increased local estrogen production, and susceptibility to breast hyperplasia and cancer. Aromatase is the dominant source in intratumoral estrogen production for postmenopausal mammary cancers.

Western Blot: Aromatase Antibody

Western Blot: Aromatase Antibody

Bogan’s group performed immunoblotting using the aromatase antibody to help them systematically profile genes expressed in the primate corpus luteum during the luteal phase of menses1. Those molecular and cellular determinations with the aromatase antibody were followed soon after by similar genomic studies from the same group, but this time they evaluated the spontaneous regression phase of menses2.  Taken together, both of these studies that used the aromatase antibody give a detailed database and profile of primate physiology. A research group from China also used the aromatase antibody in immunoblotting to investigate the role of a novel estrogen receptor (ER) variant, ER-alpha, in testosterone carcinogenesis3. Their data suggests that ER-alpha stimulates both the ERK and Akt signaling pathways. A group from Italy used the aromatase antibody to survey aromatase gene and protein levels in peripheral blood leukocytes (PBLs) and tissues4.

Novus Biologicals offers aromatase reagents for your research needs including:

PMIDs

  1. 18258683
  2. 18948396
  3. 19775474
  4. 22978812