Novus Biologicals Blog

CD56 + NCAM1 (Cluster of differentiation 56 + neural cell adhesion molecule 1)

September 29th, 2014

CD56 is a member of the Ig super family and comprises five Ig-like domains and two extracellular fibronectin-type III domains. It is expressed as three major isoforms within the nervous system, on NK cells, and a specific set of T-cells. CD56+ NK and T-cells are unique in their ability to mediate cell-mediated cytotoxicity against certain tumor cell targets without MHC restriction. Other CD56 physiological functions include: mediating cell adhesion, triggering neurite extension and migration, and brain synapse formation. CD56 is also crucial for neuronal development and plasticity in the adult brain. It is used as a tumor marker in various cancers (NK lymphomas and Merkel cell carcinoma). As another member of the Ig super family superfamily, NCAM1 consists of five extracellular Ig-like domains and two fibronectin type III domains. It has at least 20-30 distinct isoforms due to alternative splicing and sialylation posttranslational modifications.

Immunohistochemistry-Paraffin: CD56 + NCAM-1 Antibody

Immunohistochemistry-Paraffin: CD56 + NCAM-1 Antibody

A CD56 + NCAM1 antibody was initially used in Mooi’s lab in the Netherlands Cancer Institute to create histological profiles on a large bank of 350+ resected lung tumors1.  This profile typing validated the use of the CD56 + NCAM1 antibody as an identification and characterization tool for the challenging task of distinguishing between the major types of lung cancers. Related studies from the same group relied upon extensive use of the CD56 + NCAM1 antibody in profiles of normal tissues2.  These particular studies focused on determining CD56 + NCAM1 antibody reactivity in endocrine and neuron-supporting normal tissues. Studies out of Brezicka’s group employed the CD56 + NCAM1 antibody to detect and differentiate small cell lung cancer (SCLC) from other lung tumor histotypes in patients suspected to have primary lung tumors3.

Novus Biologicals offers CD56 + NCAM-1 reagents for your research needs including:

  • CD56 + NCAM-1 antibodies
  • CD56 + NCAM-1 antibody pairs
  • CD56 + NCAM-1 ELISA kits
  • CD56 + NCAM-1 lysates
  • CD56 + NCAM-1 peptides and proteins
  • CD56 + NCAM-1 RNAi

PMIDs

  1. 2837665
  2. 2832332
  3. 1654059

CD34 (Cluster of differentiation 34, hematopoietic progenitor cell antigen)

September 26th, 2014

CD34 is a cell-surface glycoprotein type 1 transmembrane protein that belongs to the sialomucin family. CD34 comprises of an intracellular cytoplasmic domain with consensus sites for serine, threonine, tyrosine and active protein kinase C (PKC). Because it is differentially glycosylated within different cell types, it has a range of apparent molecular weight sizes. It is a marker for pluripotent hematopoietic stem or progenitor cells and has been extensively used to isolate and characterize these progenitor cells. CD34 is also expressed on vascular endothelium, bone marrow stroma, embryonic fibroblasts, and neurons. As CD34-positive populations expand and differentiate into their destined lymphohematopoietic lineages, they lose their hallmark CD34 expression.

Immunocytochemistry/Immunofluorescence: CD34 Antibody

Immunocytochemistry/Immunofluorescence: CD34 Antibody

A number of malignancies (certain sarcomas and fibromas, preB-ALL, peripheral nerve sheath tumors, and papillary thyroid carcinoma) appear to also exhibit CD34-positive expression. Endothelial cell (EC) isolation studies employed flow cytometry and a CD34 antibody from Dong’s lab demonstrate a rapid and reproducible strategy for isolating murine ECs that is free of contamination1.  Zheng et al relied upon immunohistochemical with a CD34 antibody to validate their system of rapidly and reproducibly generating hair follicles from dissociated cells2. Their studies confirm that early developmental stages within an epithelial-mesenchymal system require an underlying epithelial structured platform. Sironi’s group at the Mario Negri Institute used the CD34 antibody, coupled with whole genome transcriptional profiling, to monitor their isolation and characterization of a promising endothelial cell line as a source of lymphatic endothelium3. Flow cytometry and immunohistochemistry with the CD34 antibody allowed Kalabis’ lab to identify a key stem cell population within the esophagus4. They characterized the self-renewing capacity of this particular cell cohort using a clonogenic assay and 3D organotypic culture model.  Additionally, the CD34 antibody allowed Cambridge researchers to examine the role of endothelial hypoxia inducible transcription factors HIF1 and HIF2 in metastasis regulation and malignancy5.

Novus Biologicals offers CD34 reagents for your research needs including:

PMIDs

  1. 93010164
  2. 15854024
  3. 16534603
  4. 19033657
  5. 22264788

Ovarian Cancer Infographic

September 25th, 2014

September is Ovarian Cancer Awareness month and brings to focus a cancer that is estimated to be diagnosed in over 21,000 women in the US in 2014 (1). Ovarian cancer often goes undiagnosed due to the lack of symptoms until it metastasized into the pelvic or abdominal areas. Treatment typically requires surgery and chemotherapy.

Ovarian cancer infographic

 

By: Lisa Ikariyama; Design: Kim Mesman

Download our ovarian cancer infographic

References:

  1. Cancer.org 
  2. Mayo Clinic 
  3. Ovarian.org 
  4. UCHC.edu 
  5. Cancer.gov 
  6. Cancer.gov 
  7. Cancer.net 
  8. Army.mil 

PSMA (Prostate specific membrane antigen, Glutamate carboxypeptidase II)

September 24th, 2014

Prostate specific membrane antigen (PSMA), also known as Glutamate carboxypeptidase II (GCPII), is a type II transmembrane glycoprotein that belongs to the M28 peptidase family. It acts as a glutamate carboxypeptidase on different substrates such as folate as well as the neuropeptide N-acetyl-l-aspartyl-l-glutamate. PSMA is expressed in a number of tissues including prostate, kidney, and both the central and peripheral nervous systems. A mutation in this gene is associated with impaired intestinal absorption of dietary folates, resulting in low blood folate levels and subsequent hyperhomocysteinemia. Expression of PSMA in the brain may be involved in a number of pathological conditions associated with glutamate excitotoxicity.

Western Blot: PSMA Antibody

Western Blot: PSMA Antibody

In addition, PSMA is an established cancer marker. In particular, it is upregulated in prostate cancer and has proven to be an effective diagnostic and prognostic indicator of cancer.  Labeling experiments using a PSMA antibody allowed Sacha’s group to map expression of GCPII within the human brain to better research its potential as a therapeutic target for neurotoxic glutamate-based diseases such as ALS, Huntington’s, epilepsy, and Alzheimer’s disease (AD)1.  The same group also used the PSMA antibody in follow-up domain mapping studies where they identified N- and C-termini regions responsible for enzymatic activity and proper folding2. Studies out of Rovenska’s group focused on analyzing the utility of rat and pig animal models for enzyme and in vivo disease studies3.  They employed the PSMA antibody in immunoblotting and determined that while the other species orthologs were suitable for enzyme studies, diffuse and variation expression patterns were limitations for their use in drug discovery. The PSMA antibody was used to validate the utility of performing multiplex antibody epitope mapping with a bacterial cell-surface display system4.  Additionally, tissue microarrays analyzed with the PSMA antibody allowed Jaraj et al to determine the expression of glutamate decarboxylase 1 (GAD1) in benign and metastatic prostatic tissue5. GAD1 has potential as a prostate-specific tissue biomarker.

Novus Biologicals offers PSMA reagents for your research needs including:

PMIDs

  1. 17150306
  2. 15206943
  3. 18076021
  4. 23050090
  5. 2109188

GFP – Be Green!

September 22nd, 2014

Green fluorescence protein (GFP) is a 27KD protein derived from the jellyfish Aquorea victoria that emits a green light (emission peak at a wavelength of 509 nm) when excited by blue light (excitation peak at a wavelength of 395 nm). GFP is a highly versatile protein that has become an invaluable tool in cell biology research because of its intrinsic fluorescence without substrate requirement, ability to be visualized over time durations – both short- and long-term – in living cells, and short sequence making it easy to clone and use an unobtrusive tag. GFP fluorescence is stable under standard fixation conditions and suitable for a huge variety of applications. Due to all of these advantages, GFP has been widely used as a gene expression reporter and tag, enabling researchers to visualize and localize proteins within living cells without the extra burden or complication of staining.

Immunocytochemistry/Immunofluorescence: GFP Antibody

Immunocytochemistry/Immunofluorescence: GFP Antibody

Other GFP applications include of protein-protein interaction assessments with the yeast two-hybrid system and distance measurement between proteins using fluorescence energy transfer (FRET) techniques. Usage of GFP technology has contributed considerably to a greater understanding of cell physiology, protein localization and movement within cellular compartments, and live cell trafficking and movement in cancer metastasis and normal development. There is a vast body of publications based upon use of GFP across all disciplines of biology and medicine. Immunohistochemistry experiments with the GFP antibody from Day et al were done to validate the role of the A2A adenosine receptor activation in bone marrow-derived cells as a protective measure against ischemia-reperfusion injuries within the liver1. Additional GFP antibody immunohistochemistry allowed researchers at the Mayo Clinic to monitor a novel lentivirus-based transgene expression primate eyes2.  Their work suggests this delivery system could be applied to human glaucoma gene therapy. Stem cell therapy studies used the GFP antibody to demonstrate the importance of cardiomyocyte lineage cells for stem cell therapy treatment of myocardial infarction3.  Studies out of Powell’s lab were published in Cell that highlighted the role of pan-erbB negative regulator (Lrig1) as an intestinal stem cell marker that is also a tumor suppressor negative regulator4. Immunoblotting with the GFP antibody allowed Lund et al to examine the differential effects of a variety of heterogeneous nuclear ribonucleoproteins (hnRNPs) on HIV-1 expression5.

Novus Biologicals offers GFP reagents for your research needs including:

PMIDs

  1. 15814735
  2. 19301472
  3. 19940262
  4. 22464327
  5. 22187150

Integrin beta 1 binding protein 2

September 19th, 2014

ITGB1BP2 is a muscle-specific protein cloned by a rat created by Branccio’s group in Italy that was found to interact with the cytoplasmic domain of integrin beta 11. It is expressed only in heart and skeletal muscle but is not essential for normal development and differentiation of these tissues. Branccio’s group published a follow-up study in Nature Medicine using an ITGB1BP2 antibody that the protein plays a critical role in sensing mechanical stress due to pressure overloading2. This response protects against subsequent compensatory dilated cardiomyopathy and contractile dysfunction. Further work from this group confirmed that ITGB1BP2 nucleotide variations are found in families of patients with hypertension and/or primary dilated or hypertrophic cardiomyopathy3.

Immunocytochemistry/Immunofluorescence: ITGB1BP2 Antibody

Immunocytochemistry/Immunofluorescence: ITGB1BP2 Antibody

Gu et al believe that altered ITGB1BP2 expression and signaling parallels the recovery of cardiac function (or lack of it) after myocardial infarctions (MI) and can be measured as one component in a profile panel of remodeling markers4. A recently submitted grant5 suggests that used a monoclonal ITGB1BP2 antibody for western blot to show test the functional activity of recombinant melusin. IGB1BP2 activates the downstream AKT and ERK signaling pathways by acting as a chaperone and binding to the Hsp90 machinery.

Novus Biologicals offers ITGB1BP2 reagents for your research needs including:

PMIDs

  1. 10506186
  2. 12496958
  3. 20017903
  4. 2154627
  5. US8658598 B2

Beta III tubulin

September 18th, 2014

The Beta III tubulin protein is abundantly present in both the central and peripheral nervous systems (CNS and PNS), where it is predominantly expressed during fetal and postnatal development. In cerebellar and sympathoadrenal neurogenesis, Beta III distribution is neuron-associated and present in distinct temporal-spatial gradients that are dictated by the regional neuroepithelia of origin. In addition, CNS subventricular zones consisting of neuronal and glial precursor cells exhibit transient beta III expression, where it may enable the identification of presumptive neurons derived from embryonic stem cells. In contrast, Beta III distribution is almost exclusively neuron-specific in adult tissues. Researchers published in Cell share their findings that the receptor tyrosine kinase Ret is required for motor axon attraction by integrating diffusible- and contact-axon guidance signaling in a hierarchical GPI-receptor signaling system1.

Immunocytochemistry/Immunofluorescence: Tubulin Beta 3 Antibody

Immunocytochemistry/Immunofluorescence: Tubulin Beta 3 Antibody

Their experiments with the Beta III tubulin antibody allowed them to construct a network from combinatorial components. Rudenko’s group from NIH used the Beta III tubulin antibody in their aging studies centered on the role of autosomal-dominant missense mutations on the multidomain leucine-rich repeat kinase 2 (LRRK2) that contains both kinase and ATPase activity2. They determined that the G2385R variant is a partial loss-of-function mutant in Parkinson’s disease (PD) that appears to be kinase-activating-independent. Immunoblotting and immunohistochemistry studies with the Beta III tubulin antibody performed in the Yoo lab focused on the chronic effects of pyridoxine vitamin B6 on forebrain ischemic damage and subsequent neuroblast differentiation3.  Using the Beta III tubulin antibody, Ghod’s group examined the role of the anti-inflammatory drug dimethylfumarate (DMF) on proliferation, apoptosis, and differentiation in glioma models4. They found DMF to be promising as a new treatment modality for brain tumors. The Beta III tubulin antibody allowed Kong et al to monitor the effects of chronic exposure to low doses of methylmercury on adult rat somatosensory cortexes5. Their comparative proteomic studies demonstrated the induction of a state of metabolic deficit.

Novus Biologicals offers Tubulin Beta 3 reagents for your research needs including:

PMIDs

  1. 22304922
  2. 22612223
  3. 22228142
  4. 24404403
  5. 23984759

CD19: An Undoubted Biomarker for B Cells

September 17th, 2014

CD19 is a cell surface protein member of the large immunoglobulin superfamily that complexes with CD21, CD81, and CD225 in the membrane of mature B-cells. A major function of CD19 is to assemble with the antigen receptor of B-lymphocytes to decrease the threshold for receptor-dependent stimulation, thus enhancing the specificity and sensitivity of B-cells towards antigens. CD19 plays a large role in regulating B-cell growth. Its expression is confined to only B-lymphocytes and follicular dendritic cells of the hematopoietic system. Leukemia phenotype studies suggest that CD19 is the earliest and broadest B-cell restricted marker. Because increased CD19 expression stimulates autoantibody production, CD19 studies provide insight into the autoimmunity process.

Immunohistochemistry: CD19 Antibody

Immunohistochemistry: CD19 Antibody

Defects in CD19 cause hypogammaglobulinemia. As reviewed by Gold et al, a key step in B-cell receptor (BCR) signaling involves using CD19, Cbl, Gab1 and Gab2 as docking proteins to recruit PI3K to the plasma membrane through Src homology (SH) domains1. Because most B-cell malignancies express CD19, much research is focused on new therapies for these malignancies that do not respond to standard therapies. The latest studies are focused on the development of chimeric antigen receptor (CARs) – fusion proteins that are hybrids of antigen recognition moieties and T-cell activation domains, as reviewed in Kochenderfer’s paper from the NIH2. Experiments with the CD19 antibody from Ogembo’s lab examined the regulation of RBC-pathogen clearance and filtration of altered self3. They embarked on a marker characterization of the littoral cell (LC) found within the venous sinus-lining of the red pulp within the spleen. Their profiling compared humans and closely related primates and they were able to identify SIRP alpha (CD172a) and FHOD1 as unique markers to humans.

Novus Biologicals offers CD19 reagents for your research needs including:

PMIDs

  1. 11043767
  2. 23546520
  3. 22490440

Glucose-6-phosphatase (G6PC) – A key to regulate your blood sugar level!

September 16th, 2014

The integral endoplasmic reticulum membrane-based enzyme G6PC hydrolyzes its substrate glucose-6-phosphate into glucose. Specifically, G6PC breaks down D-glucose 6-phosphate to D-glucose and orthophosphate. Because G6PC forms with the glucose-6-phosphate transporter (SLC37A4/G6PT), the resulting complex is responsible for glucose production. Thus, G6PC is the key enzyme in glucose homeostasis, functioning in both the processes of gluconeogenesis and glycogenolysis. Defects in the enzyme cause glycogen storage disease type I (von Gierke disease). Not surprisingly, G6PC is localized mainly in the liver and kidneys.  It is unique in that it is membrane-bound, unlike most other enzymes that act upon water soluble substrates.

Immunohistochemistry-Paraffin: G6PC Antibody

Immunohistochemistry-Paraffin: G6PC Antibody

An overview of the G6PC system including discussions of its regulation by glucose, insulin, cAMP, and glucocorticoids can be found in van Schaftingen’s review article1. Banka and Newman have more recently presented a clinical and molecular review on G6PC mutations and their physiological manifestations and links to various diseases2. Cicherchi et al employed the G6PC antibody to help them document the downstream effects of uric acid-dependent inhibition of AMP kinase (AMPK) in diabetes and starvation through an evolutionary standpoint3. Their data shows that an uricase mutation dating back 1.5 x 107 yrs ago to the hominid Miocene period (and triggered by a widespread famine period in Europe) when expressed in modern cell lines, likely conferred a survival advantage. This adaptation helped human ancestors to maintain critical glucose levels under situations of near-starvation, but in modern times, only serves to feed diabetes and insulin resistance.

Novus Biologicals offers G6PC reagents for your research needs including:

PMIDs

  1. 11879177
  2. 23758768
  3. 24755741