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Go Ahead! Make My DNA

April 18th, 2014

DNA methylation plays a critical role the long-term silencing of transcription and is essential for processes such as embryonic development, germline differentiation, and tissue maturation. Dnmt3a is a member of the C5-methyltransferase family that repairs cytosines in dsDNA using a unique nucleophilic attack mechanism dependent upon the transfer of a methyl group from folate intermediates onto nucleic acids.  Dnmt3a in particular performs genome-wide de novo methylation, establishes developmental DNA methylation patterns as well as paternal/maternal imprinting, and is required for methylation of most imprinted loci contained in germ cells. It can be found in both nuclear and cytoplasmic compartments and accumulates in major satellite repeats within pericentric. Canadian researchers collected compelling evidence for an essential role of Dnmt3b (but not Dnmt3a) in apoptosis of cancer cells1. The same group also published a Nature Genetics study using a Dnmt3a antibody in combination with genetic and pharmacologic inhibitors of DNA methyl transferases to better understand the role of various DNMT isotypes in oncogenic methylation2. Their studies in a wide variety of cancer cells demonstrated that Dnmt1 alone is necessary and sufficient for methylation maintenance.

Immunocytochemistry/Immunofluorescence: Dnmt3a Antibody

Immunocytochemistry/Immunofluorescence: Dnmt3a Antibody

A Dnmt3a antibody allowed Shen et al to profile changes in CpG island methylation during in vitro differentiation of human embryonic stem cells (ESCs), which enabled them to find abnormal methylation events occurring in a particular subset of genes which has implications for future cell transplant studies3. ChIP studies in conjunction with Dnmt3a antibody profiling allowed the creation of epigenetic silencing patterns in breast cancer cells with regards to two particular tumor suppressor genes of interest (caspase 9 and maspin)4. Stem cell research from the Whitehead Institute used a Dnmt3a antibody to characterize loss-of function induced pluripotent stem cells5. Interestingly, it was found that both Dnmt3a and Dnmt3b were dispensable in the nuclear reprogramming of somatic cells into a more pluripotent phenotype.

Novus Biologicals offers Dnmt3a reagents for your research needs including:

PMIDs

  1. 12015329
  2. 12496760
  3. 16870691
  4. 20132554
  5. 21576263

Apoptosis Happens

April 17th, 2014

Cell death via apoptosis is a basic cellular function occurring through the cell death receptor family and their ligands which signal through downstream adaptor molecules and the caspase protease family. Caspases have a precursor form composed of a prodomain, and large and small catalytic subunit, and are activated through a cleavage adjacent to an aspartate to liberate units and allow formation of an a2b2 tetramer. Caspase 3 is a cytoplasmic caspase with two isoforms (one acts as a dominant negative inhibitor), and is involved in the activation cascade for apoptosis execution. Caspase 3 cleaves/activates SREBPs, Caspase 7, and Caspase 9. A Caspase 3 antibody was used by Berges and colleagues to assist in their detailed characterizations of downstream proteosome inhibition pathways by the inhibitor bortezomib in an activated T-cell system1.  Studies with a Caspase 3 antibody allowed Fuzio et al to study in vivo neoadjuvant Androgen Derivation Therapy (ADT) and its effects on TGF-beta expression and signaling in prostate cancer2.  Their data suggests that TGF-beta stimulates epithelial cell proliferation and blocks apoptosis in an autocrine loop.

Immunohistochemistry-Paraffin: Caspase 3 Antibody

Immunohistochemistry-Paraffin: Caspase 3 Antibody

In exciting gene therapy studies from lysosomal storage disease (LSD) researchers in Tokyo, a Caspase 3 antibody enabled those researchers to validate the unique promise of induced pluripotent stem cells (iPS) as model systems for studying genetic LSD diseases3. Neurological studies out of Hermel’s group at the Buck Institute employed a Caspase 3 antibody in their autosomal dominant progressive disorder Huntington’s disease (HD) studies, where they were able to exclude the participation of caspases 3, 8, and 9 in selective death and pathogenesis in the neural striatum and cortex4.  Additionally, immunohistochemical studies with a Caspase 3 antibody were done in Herold’s group to allow the creation and validation of a novel extracorporeal perfusion bioreactor system using rat lingual skin folds5.

Novus Biologicals offers Caspase 3 reagents for your research needs including:

PMIDs

  1. 19735079
  2. 22269108
  3. 20385825
  4. 14713958
  5. 22223358

Different roles of CD31/PECAM1

April 16th, 2014

Platelet endothelial cell adhesion molecule 1 (PECAM1), also known as cluster of differentiation 31 (CD31), is a cell-surface glycoprotein expressed on platelets, monocytes, neutrophils, some types of T-cells and NK (natural killer) cells. It makes up a large portion of the endothelial cell intercellular junctions. CD31/PECAM1 is a member of the immunoglobulin superfamily and plays many different roles involving leukocyte migration under most inflammatory conditions, angiogenesis, integrin activation, atherosclerosis and thrombopoiesis.

Diseases associated with CD31/PECAM1 include angiosarcoma and pulmonary vein stenosis.  Among its related super-pathways are response to elevated platelet cytosolic Ca2+ and hemostasis.

CD31/PECAM1 is commonly used as an endothelial marker, particularly in angiogenesis. It has an important role in assisting placental and endometrial vascular development. However, it is also associated with many negative angiogenic occurrences, particularly those within or related to tumors and other disease lesions.  These include MS, cardiac angiosarcoma and prostate cancer.

Novus CD31/PECAM antibody NB600-562 was used in a study about quantification and prognostic relevance of angiogenic parameters in invasive cervical cancer. Tumor stromal neovascularization was investigated in carcinomas of the uterine cervix by staining representative sections with the specific endothelial marker anti CD31 (clone JC/70A, isotope IgG1). It has been shown that all angiogenesis parameters had prognostic value. In the future these criteria may be used for selection of patients for anti-angiogenesis therapy. (1)

Western Blot: CD31/PECAM1 Antibody (JC/70A) [NB600-562]

Western Blot: CD31/PECAM1 Antibody (JC/70A) [NB600-562]

CD31 is thought to contribute to the physiological regulation T cell homeostasis due to the presence of two immunotyrosine-based inhibitory motifs in its cytoplasmic tail.  Loss of CD31 expression leads to uncontrolled T cell-mediated inflammation and certain CD31 polymorphisms correlate with increased disease severity in human graft-versus-host disease and atherosclerosis. Kishore’s group used the Novus CD31 antibody NB100-92205 in their study proposing that CD31-mediated modulation of memory T cell chemokinesis is a key mechanism by which this molecule contributes to the homeostatic regulation of effector T cell immunity. (2)

PECAM-1 (CD31) might also play an important role in regulating antigen-specific T cells trafficking in CNS inflammatory diseases. (3)

  1. PMID: 9683289
  2. PMID: 22724015
  3. PMID: 11487054

Novus Biologicals offers CD31/PECAM1 reagents for your research needs including:

Osteoprotegerin: The Bone Protector

April 14th, 2014

Osteoprotegerin (OPG) is a secretory glycoprotein that is a family member of the TNF receptor (TNFR) superfamily. Osteoprotegerin protects bone by blocking osteoclastogenesis and increasing bone density. Unlike other TNFRs, osteoprotegerin lacks a transmembrane domain as well as any apparent cell-associated signals. High levels of osteoprotegerin mRNA are found in specialized tissues such as lung, heart, kidney, and placenta. There is evidence that osteoprotegerin plays a role in degenerative arterial disease. Sandra et al used ELISA studies employing the osteoprotegerin antibody with hopes of understanding the bone resorption that occurs with the bone tumor ameloblastoma (1). Their findings seem to suggest that ameloblastoma secretes RANKL and TNF alpha. Some interesting studies looking at bone loss in inflammatory bowel disease (IBD) used the osteoprotegerin antibody to establish a murine model of colitis-associated bone loss, and to demonstrate that IBD is associated alterations in the RANKL and OPG system2.  Larson’s group used the osteoprotegerin antibody to create a detailed analysis of bone remodeling proteins associated with the osteoblastic and osteolytic response in human prostate cancer bone metastases3.  The osteoprotegerin antibody enabled Australian researchers to determine that levels of both OPG and osteopontin (OPN) are upregulated in carotid atherosclerosis4.

Immunohistochemistry-Paraffin: Osteoprotegerin Antibody

Immunohistochemistry-Paraffin: Osteoprotegerin Antibody

Pettit et al used osteoprotegerin antibody in their immunohistochemical characterizations of RANKL expression at the interfaces of articular bone erosion in human rheumatoid arthritis samples5. These researchers found that the resulting expression patterns of bone remodeling factors such as RANK, RANKL, and OPG creates a microenvironment highly conducive to osteoclast differentiation and activity.

Novus Biologicals offers Osteoprotegerin reagents for your research needs including:

PMID

  1. 15935726
  2. 15753532
  3. 23334979
  4. 15143295
  5. 016490750

KLF4 opens the door for stem cell research

April 11th, 2014

KLF4 (Kruppel-like factor 4, Epithelial zinc finger protein EZF) is a zinc finger transcription factor thought to be involved in developmental differentiation and proliferation. It is considered a pluripotency reprogramming factor (PRF) due to its ability to change cell fate via gene expression conversion. Other PRFs including Sox2, Oct4 and KLF4, as well as a discussion about their capabilities, are reviewed by Jauch et al in hopes of enabling the engineering and optimization of PRFs1. Interestingly, recent studies suggest KLF4 may play a role in vascular disease due to its expression in multiple vascular cell types, but this remains to be seen2. Huang’s group used a KLF4 antibody to examine the first generation conversion of quiescent bovine cells into induced pluripotent stem cells (IPSCs) with a novel virus-free poly-promoter vector3.

Immunohistochemistry-Paraffin: KLF4 Antibody

Immunohistochemistry-Paraffin: KLF4 Antibody

They were able to initiate a self-sustaining pluripotency program unlike any seen in a human system. Likewise, a KLF4 antibody enabled researchers from Temple University to test their hypothesis that the hepatitis B antigen HBx triggers transformation through promoting properties characteristic of cancer stem cells (CSCs)4. Their studies found that through the use of a KL4 primary antibody that HBV does indeed promote stem-cell associated properties in hepatocellular carcinoma (HCC) by mimicking CSCs.

Novus Biologicals offers KLF4 reagents for your research needs including:

PMID

  1. 23642061
  2. 24573018
  3. 21912700
  4. 21464043

Prostate Cancer Infographic

April 8th, 2014

Prostate cancer is caused by malignant cells developing in prostate tissue. Common warning signs of prostate cancer include problems with urination (sudden urges, pain, blood in urine, difficulty urinating), experiencing pain in the back and pelvis, and feeling tired/dizzy. There are different tests utilized to diagnose prostate cancer including PSA screening, TRUS, DRE, and biopsy.

Prostate Cancer Infographic

Resources:

  1. Cancer.gov
  2. Cancer.gov 
  3. CDC.gov 
  4. CDC.gov 
  5. Cancer.org 
  6. PCF.org 
  7. Movember.com

By: Lisa Ikariyama; Design: Cheryl Reyes

Breast Cancer Infographic

April 4th, 2014

Breast cancer is caused by malignant cells developing in breast tissue. It is the most commonly diagnosed cancer in women, but advancements in treatment options have seen the death rate decline since the 1990s. Common warning signs of breast cancer include lumps, changes in breast size or shape, discoloration, dimpling of the skin, new concentrated pain in the breast, and rash on the nipple.  Yearly mammograms and self-exams are an important part of early detection of breast cancer.

Breast Cancer Infographic

     Download our breast cancer infographic

Resources:

  1. National Breast Cancer  
  2. Susan G Komen
  3. Cancer.gov 
  4. SDSU.edu 
  5. Komen Kansas City
  6. CDC 
  7. Cancer.gov 
  8. Breast Cancer Awareness 

By: Lisa Ikariyama; Design: Cheryl Reyes

 

CXCR7 chemokine is not kind: Spotlight on proinflammatory chemokine receptor type 7

April 3rd, 2014

The CXCR7 (C-X-C chemokine receptor type 7) proinflammatory protein is a member of the G-protein coupled receptor (GPCR) family. It is a transmembrane protein first identified as the EBV-induced gene-1, and while it was originally classified as an orphan receptor, it is now known to be a novel and alternate receptor for the chemokines CXCL11 and CXCL12. While the actual function of the CXCR7 protein is not entirely understood, it appears to be elevated in human cancers and important for tumor proliferation, invasion, and metastasis. In particular, the CXCR7 ligands have abundant expression in human astrocytoma and glioblastomal neural tissues.  Singh’s oncology group used a CXCR7 antibody in their immunoprecipitations to identify a novel pathway of ligand-independent signaling in prostate cancer that involves paired regulation with IL-81. Choy et al also used a CXCR7 antibody in their T-cell humanized mouse model, developed around arteric allograft rejection2. There, they found that CXCL12 induces nitric oxide (iNOS) in a CXCR4, but not CXCR7, dependent fashion.German researchers tested a CXCR7 antibody in both astrocytes and Schwann cells to confirm that CXCR7 is necessary in both cells types for CXCL12-dependent mitogenesis3. Further neural tissue studies in Hattermann’s lab with a CXCR7 antibody determined that CXCR7 mediates resistance to drug-induced apoptosis by agents such as camptothecin and temozolomide4. Using a primary antibody to CXCR7, or any of the receptors in the CXCR7 family, has proven successful in understanding the dynamic qualities of this protein.

  1. 21398406
  2. 19059114
  3. 20197403
  4. 20388803

Novus Biologicals offers CXCR7 reagents for your research needs including:

Top 10 Things Only People in a Lab Will Understand

April 1st, 2014

After working on several thousand experiments to test products, rigorously quality testing data, and validating products in the lab at Novus Biologicals, we have developed a list of things that only scientists will understand from spending time in a lab. Happy April Fools’ Day and enjoy our top 10 list!

Ten things only a person in a lab will understand

 

By: Amelia Zommer, Sam Garcia, Andrew Cosgrove and Lisa Ikariyama

Beta Actin Antibodies: Much More than a Loading Control

March 31st, 2014

Beta-actin belongs to a large family of highly conserved structural cell proteins that regulate cell motility, structure, and integrity. Beta-actin is expressed in all eukaryotic cells making it the ideal internal quantitative control for protein comparative assays. This feature has made it uniquely a historical and heavily-utilized standard, as the public record of scientific publication literature can attest to. Ex vivo familial genome-wide genetics studies trying to identify chemotherapy cytotoxicitygenomic hotspots used the beta actin antibody in genome-wide linkage analyses1. Dietz’s group employed the beta actin antibody to monitor effects of long-term administration of the DNA synthesis inhibitor imatinib meslyate on downstream events such as proliferation and delayed-type hypersensitivity (DTH) in both human primary T-cells and murine in vivo models2.

Immunocytochemistry/Immunofluorescence: Beta Actin Antibody

Immunocytochemistry/Immunofluorescence: Beta Actin Antibody

In the neurodegeneration arena, Park et al characterized atherosclerotic plaques in ApoE knockout mice with the beta actin antibody to analyze the role of ectopic cell cycle activation in classic pathological hallmarks found in Alzheimer’s such as plaque pathogenesis and rupture3. Antibodies to beta-actin itself also allow scientists to study and understand the cytoskeleton and structural components within the cell required for basic movement and cell functions. In a series of experiments, Gimona’s group generated a beta actin antibody and elegantly showed that it decorates actin filaments as well as peripheral lamellipodia4. High resolution dual-label immunocytochemistry studies with the beta actin antibody localized the thin filament-binding protein calponin to both the contractile apparatus and cytoskeleton in smooth muscle cells5. The beta actin antibody was used in lipid metabolism studies assessing metabolic transformations driving breast cancer cell growth and survival6.

Novus Biologicals offers Beta Actin reagents for your research needs including:

PMIDs

  1. 15282376
  2. 15100154
  3. 17360920
  4. 8162619
  5. 8006064
  6. 24070020