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Posts Tagged ‘Antibody’

The Use of Hypoxia Antibodies in Stem Cell Cultures

Tuesday, April 27th, 2010

Stem cell antibodies are raised against specific stem cell marker proteins, which are used to identify and isolate stem cells in vitro. We at Novus Biologicals have over 6000 products on our antibody database specific to adult and embryonic stem cell research.

Whereas adult stem cells can only differentiate into one cell type, embryonic stem cells (ESCs) have the unique ability to differentiate into a large variety. Stem cells can be genetically manipulated into specific cell lines. A recent development was the creation of ESC-like cell lines from adult stem cells.

Stem cells are found in both healthy and diseased tissue. Cancer stem cells (CSCs) are expressed in both solid tumours and leukaemias, and are of particular interest as they are resistant to many therapies, and are thought to drive metastatic spread of cancer cells. Hypoxia is closely linked to CSC research, as disruptions of hypoxic pathways are linked to anti-apoptosis and the development of cancer cells. Stem cells are often found in hypoxic environments in vivo.

This has relevance in the culturing of stem cells for in vitro assays, since low oxygen pressures can affect cultures in a number of ways. This includes decreased spontaneous differentiation rates, and a reduction in the dedifferentiation of tumour cells being generated for cancer cell lines. Hypoxia can also improve the generation rate of pluripotent stem cell generation.

Thus, controlled hypoxia in stem cell culture can be seen as beneficial. However, at present it is difficult to control oxygen levels to precise levels, and therefore hypoxia markers must be utilised.

At Novus Biologicals, we have an extensive antibody catalogue, with many specifically for embryonic stem cell research.

Routine Antibody Screen Uncovers Exciting New Cancer-Cleaning Enzyme

Friday, April 23rd, 2010

We at Novus Biologicals have a large enzyme antibody database. Enzyme antibodies are widely used in apoptosis protein tagging studies. Disruption of the proteins regulating apoptosis is known to lead to formation of tumour cells in certain cancers.

Often, the most basic of protocols can throw up surprising results. Recently, a seemingly basic antibody study into the mechanism of action of WT1 (the Wilms tumour oncogene) revealed that the protease HtrA2 may have a hitherto unknown role as a ‘cell cleaning’ enzyme, able to eradicate WT1, thus preventing development of cancer.

The Wilms tumour is an embryonic cancer identified by mutation of WT1, a transcription protein that normally acts as a tumour suppressor during kidney development. WT1 has several transcript variants and can cleave in several ways to bind to other cellular factors. It is expressed at high levels in a number of childhood and adult cancers, including myeloid leukaemia, and breast and lung tumours.

WT1 has been extensively studied, although its role in the development of non-Wilms cancers is poorly understood. We at Novus Biologicals have a large number of WT1 products in our antibody catalogue, allowing researchers to delve into binding mechanisms at WT1 domain sites. It was while conducting such a trial that Doctors Joerg Hartkamp and Stefan Roberts, of Manchester University, discovered the cell-cleaning attributes of HtrA2. Previously known only to play a role in programmed cell death, it was found to remove WT1 from cancer cells.

HtrA2 antibody research has been conducted as far back as 1996. This exciting news proves there is always something new to be found, on even the oldest antibody database.

Antibody Research into Rare Tumours could Lead to Cutting-Edge Cancer Treatments

Thursday, April 22nd, 2010

We at Novus Biologicals have a vast number of products in our antibody catalogue, targeted at cancer research. They cover areas as diverse as apoptosis and cell signalling pathways, and antibodies include those derived from a number of tumour cell lines.

Now, it looks as if our cancer antibody database is set to expand once more. Scientists have recently uncovered exciting new facts on sarcomas – rare tumours affecting only a small minority of people, which are found in tissues throughout the body. Despite the rarity of sarcomas, the findings have enormous ramifications on the field of cancer therapy as a whole. They could pave the way for a whole catalogue of new treatments for the most common cancers affecting man.

Sarcomas are found in muscle, nerves, joint tissues, deep skin tissues and blood vessels. The proteins causing them undergo unique, well-characterised molecular changes which make them ideal models for the development of new cancer therapies. The findings have recently been made public at the 2010 ESMO conference on Sarcomas and Gastro-IntestinalStromal Tumours (GIST).

We at Novus Biologicals have over 130 antibody products targeted at sarcoma proteins which have been researched for a number of years. However, it has only been comparatively recently that novel treatments have started to emerge. Much of this is down to improvements in antibody assay techniques, enabling precise identification of protein mutations at a molecular level. The ESMO conference concentrated on the molecular alterations that occur in soft-tissue sarcomas, in particular GIST, and the development of treatments to combat them.

Bestrophin Research Reveals Potential Target for Glaucoma Therapy

Wednesday, April 21st, 2010

The Bestrophin 1 to 4 group of proteins are membrane-bound globulins, encoded by the BEST genes 1 to 4. We at Novus Biologicals have a range of products for Best research on our antibody database; mainly targeted to Best-1.

Bestrophin antibodies are routinely used in tagging studies for retinal epithelial cells, as well as ocular disease research; in particular, macular degeneration caused by mutations of Best-1. This protein, which is encoded by the BEST1 (also known as VMD2) gene, is found in the basolateral retinal membrane where it aids regulation of voltage-dependant (anion) Calcium channels. It plays an additional role in ocular development. Over 120 mutations of Best-1have been recorded. They are responsible for a wide range of ocular disease phenotypes, the most publicised of which is Best vitelliform macular dystrophy.

Recently, antibody research on another Bestrophin protein, Best-2, has suggested mutation may play a therapeutic role. The Best-2 gene encodes an anion channel in the plasma membrane of non-pigmented retinal epithelial cells, thus it plays a similar role to Best-1. Studies showed that disruptions to the Best-2 protein structure caused a reduction in intraocular pressure in mice, due to alterations in aqueous drainage and flow.

In studies conducted by Zhang et al, antibody suppliers provided rabbit polyclonal antibodies targeted to human Best-2 antigen. Human donor eye tissues were used to examine expression of human Best-2. The immunoassay results indicated that human Best-2 was expressed only in the non-pigmented epithelial cells, and may reduce intra-ocular pressure in humans in a similar way to that seen in mice. This suggests it may prove useful in the development of novel glaucoma therapies.

Mutations in CHD7 Provide Link Between CHARGE syndrome and Cancer Cell Metastasis

Tuesday, April 20th, 2010

Antibody studies into human CHARGE syndrome have shown mutated CHD7 plays a major role in its development. CHARGE originates in neural crest cells (NCCs) during early embryonic development. It leads to a number of birth defects including craniofacial, heart, brain, urogenital, hearing and growth defects. New research conducted by Wysocka et al, of the Stamford University School of Medicine, indicates a connection with tumour development in adults.

Chromatin is the tightly-packed form of DNA found within the cell nucleus. It is controlled by a number of chromatin remodeler proteins, one of which is CHD7. CHD7 acts on nuclear proteins to allow activation or deactivation of various genes.

NCCs are unusual in that they migrate freely during embryonic development, differentiating into a range of cells including neural, heart, cartilage and bone cells. Wysocka et al, of the Stamford University School of Medicine, recently conducted antibody assays exploring the critical role CHD7 plays within neural crest cells.

Previous studies had showed mutated CHD7 was expressed in CHARGE cells. By blocking CHD7 release in tadpoles, cell migration was halted and symptoms similar to those of CHARGE sufferers were expressed, proving that uncorrupted CHD7 is essential to normal embryonic development.

Further antibody studies showed CHD7 worked in combination with the PBAF chromatin remodelling complex, to control remote genes. Previous studies have shown that Slug and Twist – two genes controlled by CHD7 and PBAF – are oncogenic.

We at Novus Biologicals have a number of proteins on our antibody database, which will help further research into this area.

City Gym Helps Sheffield University in FTO Fat Gene Research

Monday, April 19th, 2010

The recently discovered FTO (Fat mass and obesity orientated) gene is of great interest to antibody research groups, throwing clearer insight into the reasons why some people have difficulty losing weight, even when following a healthy lifestyle.

Studies into serotonin (5-HT) are equally important, and its importance has been realised in a number of antibody studies into causes of weight gain. 5-HT receptors are located in the hypothalamus, which is a primary expression site for FTO. A number of research groups are currently looking at the way these two proteins may interact.

Recently, more than 70 obese volunteers signed up to a Sheffield sports centre to take part in a genetic study being conducted by the city’s Hallam University. The research, which started early in 2009 and is ongoing, looks at changes in participants’ motivation, compared to the levels of serotonin that is naturally released.

In vitro FTO antibody studies, using sera taken from human volunteers, have previously shown that higher levels of physical activity blocks the effects of the FTO gene. At the University of Maryland in 2008, researchers analysed activity levels against gene variants in a number of exercisers. They found the least FTO activity in those who were physically most active.

Serotonin, otherwise known as the ‘happy hormone’ has been shown to aid weight loss in a number of ways, including appetite suppression and motivation to energise. Therefore, it has an important role to play in blocking the release of FTO. We at Novus Biologicals have a wide range of FTO and serotonin antibodies on our antibody database, which will help to facilitate this research.

IDH1 Mutation Throws Door Open on AML Research

Friday, April 16th, 2010

We at Novus Biologicals have a large antibody database for cancer research. However, AML, or acute myeloid leukaemia, is one of the cancers for which the number of antibodies is still fairly small, owing to limited knowledge of the genes involved.

An exciting new discovery looks set to change all that. Recently, Thompson et al of Abramson Cancer Centre, Pennsylvania University, discovered mutated IDH1, or isocitrate dehydrogenase 1, in stock AML cells. The biomarker was 2HG (2-hydroxyglutarate), a metabolite specific only to the mutated form of IDH1. The results indicated that mutated IDH1 could account for up to 50% of all AML cases where a gene had previously not been identified.

In its non-mutated form, IDH1 plays an important role in the tertiary stage of the citric acid cycle. However, when mutated it acquires a novel metabolic function that results in the release of 2HG. Although both 2HG and mutated IDH1 had been shown to be overexpressed in tumour cells, it was only in November 2009 that the role of mutated IDH1 as an oncogene was ascertained, when 2HG was identified as a metabolite, and therefore biomarker for the gene in brain tumour studies. Prior to this, IDH1 was thought to have a non-functional role in tumour formation.

Cancer metabolism, in which antibody assays concentrate on the metabolic changes that take place in cancer cells, is an exciting new area that has enormous implications in the development of novel therapeutic compounds. This latest news has opened up new avenues of exploration for antibody suppliers like us at Novus Biologicals.

Amish Genetic Purity Shown to be Ideal for In Vitro Metabolic Research

Wednesday, April 14th, 2010

Human derived antibodies are widely used in disease research. These tend to be associated with cancer cell lines, such as the MCF7 breast tumour line, for which we provide both antibodies and whole cell lysate. The cells that are generated are identical in every respect to the originals, meaning reproducible results can be obtained years after the original cell lines were harvested.

When it comes to in vivo studies, however, it is generally accepted that genetically similar results are only obtainable from genetically homogenous laboratory strains (Zucker rats, ob-ob mice etc.) The idea of a genetically homogenous human strain would seem to be unfeasible, but one does, in fact, exist, in the Old Order Amish people of North America.

The Amish are a pure-bred race who can trace their ancestry back to a small group of 18th century Europeans. Their mode of living has remained remarkably unchanged since then, giving them a degree of genetic purity that is on par with a laboratory animal model, and means comparable antibody results can be obtained from a number of individuals.

The University of Maryland has been using Amish volunteers since the 1990s to investigate human clinical conditions such as diabetes, heart disease and obesity. A recent study, published in 2008, looked at the effect of increased activity on the FTO ‘fat’ gene. Despite their active lifestyle, Amish people are as obese as other Caucasian Americans.

The products we at Novus Biologicals have on our antibody database are mainly used for human research. The Maryland Amish data forms a unique contribution to this.

Overview of Signal Transduction Antibodies

Monday, April 12th, 2010

At Novus Biologicals, we have over 9,000 products on our transduction antibody database. Signal transducers are proteins that facilitate movement of signals, i.e. molecules that cause a reaction, from outside to inside the cell, where they cause changes to cellular function and activity in either the cytoplasm or the nucleus. Transduction pathways are usually activated by complex enzyme cascades, and new proteins are constantly being developed. Thus, our antibody catalogue is constantly being updated.

There are three classes of cell signalling proteins, differentiated according to the methods of transduction. The first group, steroid hormone transducers, diffuse across the cell membrane, binding to receptors within the cellular matrix. Upon binding to their ligands, these receptors are released from binding proteins, such as HSP90, via a conformational shift. A nuclear localisation sequence is unlocked, which facilitates transport of the hormone into the nucleus. Once there, the molecules bind to DNA transcription proteins and alter gene expression.

The second group, protein hormones and growth factors, are too large or not hydrophobic enough to diffuse through the membrane. In this case, transduction takes place by binding to cell-surface receptors. The entire ligand/receptor unit is then adsorbed into the cell by endocytosis. The ligand is released while the receptor is recycled or broken down.

The most common type of cell signalling is transduction via secondary messengers. Here, the ligand binds to the primary messenger (the receptor) causing a conformational change. This in turn alters the functions and/or properties of the target cell.

Disruption of all three types of transduction pathways is indicated in a number of tumours, so our transduction antibody database is essential for onconogenic research.

Weekly Highlights of Novus Antibody Lab

Wednesday, March 31st, 2010

The end of March has proven to be a busy time for the Novus antibody lab. Seven new antibodies were launched, numerous antibody conjugations completed, and continuous quality control testing of new antibody lots.

Two of Novus’ newly released products are c-Myc antibodies: clone 9E10 and clone 9E11. c-Myc is a transcription factor that is activated in a variety of tumor cells, and plays a critical role in cellular proliferation, differentiation, apoptosis and cell cycle progression. It is also a commonly used epitope tag engineered onto the N- or C-terminus of a protein of interest so that the tagged protein can be analyzed and visualized using immunochemical methods. Clone 9E11 is made to a synthetic peptide (AEEQKLISEEDL) conjugated to KLH, and is also available conjugated to Biotin, HRP, DyLight 488 and DyLight 549. Clone 9E10 is also available conjugated to Biotin, HRP, DyLight 488, DyLight 549 and DyLight 649.

Novus also released a RNA polymerase II Antibody (clone 4H8), PCNA Antibody (clone PC10), VEGF Antibody (clone VG1) and Cytokeratin 1 Antibody (clone LHK1). All new products added to the Novus antibody catalog within the past month can be viewed here.

The Novus antibody lab team conjugated several antibodies over the past week. The PMP22 Antibody (clone Hu1) was conjugated to biotin, and Blimp-1 Antibody (clone 3H2-E8) and Carbonic Anhydrase IX Antibody were both conjugated to HRP, to name just a few. A new lot of Novus’ ABCG1 Antibody (cat # NB400-132) was QC tested in Western blot on Raw 264.7 Whole Cell [T0901317 treated and untreated lysates] and a new lot of the TYRO3 Antibody was tested on mouse brain lysate achieving a clear band at 96kDa. Continual QC testing of new antibody lots assures that all of Novus’ antibodies are of the highest quality and specificity.