Murine Ba/F3 Cell Line Is Helpful to the Selectivity of Kinase Inhibitor

Kinase inhibition is an area of significant interest across academia and pharmaceutical industry. However, discovering the full range of intracellular targets of a small molecule kinase inhibitor can be a daunting task. This information is needed to understand the mechanistic basis for potential toxicities and find out the particular inhibitor which tumors may respond to.

Besides the 518 known kinases encoded in the human genome, there are over 2,000 other nucleotide-dependent enzymes, including polymerases, chaperones, motor proteins, reductases and methyltransferases, possibly containing binding sites. Ideally, the selectivity of a new kinase inhibitor is evaluated at the level of the protein, cell and whole organism.

Cellular selectivity of kinase inhibitors can be evaluated by using panels of cell lines that are engineered to report on inhibition of a particular kinase or using unbiased proteomics approaches, and many such cell lines have been developed from the murine Ba/F3 cell line.

Murine Ba/F3 cell line is pro-B-cell line whose proliferation normally requires the cytokine interleukin 3 (IL-3). However, its transformation by an oncogenic kinase will lead to IL-3-independent proliferation and survival, and thus the rate of cell growth and proliferation can be used as a read-out of intracellular kinase inhibition. Further, cytotoxicity that results from specific kinase inhibition can be discriminated from non-specific cytotoxicity through the introduction of IL-3. Panels of kinase-transformed Ba/F3 cells can be created by using either naturally occurring oncogenic fusion proteins or by creating artificial fusions between kinase domains and domain Tel found in several naturally occurring fusion proteins with a strong ability to multimerize fused proteins.

Creative bioarray offers high quality murine Ba/F3 cells or cell line for research use. Its IL-3 dependent murine pro-B-cell line is found with a lymphatic back ground. Performing May-Grünwald-Giemsa staining provide the evidence that those cells are derived from C3H mouse.

You can get murine Ba/F3 cells at

http://www.creative-bioarray.com/BA-F3-CSC-C2045-item-1604.htm

Altered Milk Protein Can Be Used to Cure Infants with AIDS

Source: biotechnology.com

A recent research done in Penn State's College of Agricultural Sciences reports that milk powder containing Ritonavir can be used as a transport system to deliver drug that is not very soluble in water.

Creative Bioarray

According to Harte, one of these researchers involved in this research, an altered protein with an antiretroviral drug in milk promises to greatly improve treatment for infants and young children suffering from HIV/AIDS,

The World Health Organization has reported that there are an estimated 3.4 million children living with HIV/AIDS, and 1/9 of them live in countries and regions where effective antiretroviral treatments still are not widely accessible or available. Most of those children cannot receive a proper treatment due to limited resources.

Another problem is that most antiretroviral drugs are not well tolerated by very young children. Ritonavir, as the most commonly prescribed antiretroviral drugs for treating and preventing HIV has undesirable side effects and important oral-delivery problems.

Ritonavir is with high hydrophobicity and low solubility in water, which leads to insufficient bioavailability. Besides, the liquid formulation to deliver Ritonavir not only tastes bad, it can also cause side effects such as nausea, vomiting and diarrhea. For those reasons a new alternative pediatric formulation of Ritonavir with better water solubility and oral flavor is needed.

Harte’s team has recently found a protein named caseins in cow's milk and this protein is said to be responsible for the white color of milk. They find that these casein micelles are able to carry molecules that have very little solubility in water with low molecular weight and could be bound to triclosan- an antimicrobial used in deodorants and vitamin D, which is added regularly to skim milk. This finding shows that casein micelles are natural delivery systems for amino acids and calcium from mother to young.

“We don't have the data yet- we find that the Ritonavir is being adequately delivered by the protein in milk. So if that works, I think we are pretty close to having a formulation that can be used with hydrophobic drugs." Harte said.

He also noted that he has made a proposal for research funding and hoped that they could finalize with a new formulation to help those children suffered from AIDS.

Their study has yet not entered the trail stage and further works are needed.

Easier Hybridization with E-308 Hybridization System

In biology, hybridization refers to the interaction of complementary nucleic acid strands. Since DNA is a double-stranded structure held together by complementary interactions (in which C always binds to G, and A to T), complementary strands favorably reanneal or “hybridize” to each other when separated. Hybridization can occur between two DNA strands and also between DNA and RNA strands provided there is sufficient complementarity in their base sequence. DNA denaturation, also called DNA melting, is the process by which double-stranded deoxyribonucleic acid unwinds and separates into single-stranded strands through the breaking of hydrophobic stacking attractions between the bases. The process of DNA denaturation can be used to analyze some aspects of DNA and detect sequence differences between two different DNA sequences. Both terms are used to refer to the process as it occurs when a mixture is heated, although "denaturation" can also refer to the separation of DNA strands induced by chemicals like urea.

Hybridization occurs in all physiological DNA reactions including replication, and transcription, and it forms the basis of many molecular biology techniques including Southern and Northern blotting, PCR and sequencing and so on. This technique has been applied to many aspects of human life, especially for biochemical and clinical research fields.

Creative Bioarray’s E-308 Hybridization system is one of the most advanced equipments in molecular biology. With programmable and humidifying system, it automates the steps in a slide-based FISH procedure, and provides walk-away convenience for clinical and research personnel. It is also very easy to operate. For example, up to 12 slides can be easily added or removed with one hand. Besides, its two-side heating of slides allows for achieving exact and prompt correspondence to the set and actual temperature and maintaining uniform temperature across all slide positions.

Features of E-308 Hybridization system：

Ÿ   Touch screen allows for easy to read, program and use

Ÿ   It can run In-situ PCR experiments

Ÿ   Possibility to export cycles data via USB memory

Ÿ   Four operation modes: denaturation/hybridization, hybridization fixed temperature, custom, In-situ PCR

Ÿ   Can be used as a fixed temperature slide warmer

Ÿ   Rapid temperature ramp-up and accuracy of ±1°C

Ÿ   Superior temperature uniformity accross all slide positions

Ÿ   Perfect humidity control system

About us

Creative Bioarray is a well-recognized biotech company propelled by integrity, expertise and the desire to contribute to simplify, improve, and accelerate research speed of the scientists by providing the highest standard quality research products and services. It provides products such as human tissue , animal tissue, tissue array, microorganisms, cells, probes and services like cell services, biosample services, histology services.

More information at http://www.creative-bioarray.com/Equipments/E-308-Hybridization-system.htm

IHC Technique: the Combination of Immunology and Histochemistry

IHC or Immunohistochemistry usually means the process of detecting antigens in cells of a tissue section by exploiting the principle of antibodies binding specifically to antigens in biological tissues. Immunohistochemical staining technique is widely used in the diagnosis of abnormal cells such as those found in cancerous tumors. Immunohistochemical staining technique featured with its high sensitivity and specificity, combining the morphological changes, functional and metabolic changes, can directly detect the existence of some proteins and polypeptides in tissue section, cell smear or cell climbing slice and can be accurate to the subcellular structure. It can quantitatively analyze the tested materials by integrating with computer image analysis system or laser scanning microscopy techniques.

History

The principle of IHC has been discovered since the 1930s, but it was till 1942 that the first IHC study was reported and introduced to the world. In 1942, a scientist named Coons et al firstly published works on IHC. His publication describing an immunofluorescence technique for detecting cellular antigens in tissue sections marked the beginning of immunohistochemistry. Since then, improvements have been made in protein conjugation, tissue fixation methods, detection labels and microscopy and IHC has now become a valuable tool in both diagnosis and research of infectious and neoplastic diseases in a variety of animals.

Application

IHC is used for disease diagnosis, drug development and biological research. With specific tumor markers, physicians use IHC to diagnose a cancer as benign or malignant, determine the stage and grade of a tumor, and identify the cell type and origin of a metastasis to find the site of the primary tumor. IHC is also used in drug development to test drug efficacy by detecting either the activity or the up- or down-regulation of disease targets. IHC is also widely used in basic research to understand the distribution and localization of biomarkers and differentially expressed proteins in different parts of a biological tissue.

Samples are prepared on individual slides, or multiple samples can be arranged on a single slide for comparative analysis, such as with tissue microarrays. IHC slides can be processed and stained manually, while technological advances now provide automation for high-throughput sample preparation and staining. Samples can be viewed by either light or fluorescence microscopy, and advances in the last 15 years have improved the ability to capture images, quantitate multiparametric IHC data and increase the collection of that data through high content screening.

About us

Creative Bioarray is a world provider of biochemical and biotechnological products and services, and it has provided high quality IHC data to research communities for many years. It has established well-tested SOPs in optimizing IHC staining conditions, which include different combinations of various tissue fixation processes, antigen retrieval conditions, antibody titration options, incubation time and washing stringency. It also has unparalleled bioinformatics support in performing IHC studies to facilitate research and product development.

http://www.creative-bioarray.com/Services/Immunohistochemistry-IHC.htm

Embryonic Stem Cells Are Put In Trail for Curing Diabetes

Source from Bioscience.com

Lately, the two teams San Diego’s ViaCyte and Boston’s Harvard University successively announced their progress on embryonic stem cells for curing diabetes, which was suggested to be the beginning of the golden age of stem cell therapeutics.

Last week ViaCyte’s announcement of their first FDA-approved embryonic stem (ES) cell clinical trial for diabetes was followed by Harvard University’s new report declaring beta cells made from ES cells “cured” diabetic mice, on which media commented: “if Harvard took it further in the dish, then ViaCyte took it further to the clinic”.

After 15 years of work, systematically testing 150 different combinations of more than 70 compounds involved in pancreas development, the Harvard team finally created mature islet cells— from both human ES cells, and human induced pluripotent stem cells. Although those ES and iPS cells are robust and can make billions of copies of themselves, they need to be placed in a yet-to-be-devised capsule, to protect them from the immune system, and contain them should they go awry.

Harvard’s’ competitor ViaCyte, which is located in the opposite site of the country, noted that they started to treat diabetes with human ES cell-derived islets more than 10 years ago and they published their method for making islet cells nearly eight years ago, and also they worked for many years on designing a device to deliver the cells to the body that would allow them to thrive and not be rejected as foreign cells.

Recently they received FDA approval to implant their cells, in patients, in a teabag-like implantable device. They will transplant the cells at a more immature stage than Harvard’s, as they found they “worked better” if allowed to mature post-transplant, in the bodies of lab animals.

However, Harvard’s team leader Dough Melton refuted that while his cells “cure” mice in days, ViaCyte’s cells “cure” mice in a few months, as they need the host metabolism to finesse differentiation and persuade the cells to respond to glucose.

The competition on stem cell research is quite fierce and there are other teams like BetaLogics, which also published article four weeks ago describing similar pre-clinical success with ES cells for diabetes.

It is really exciting news for diabetic patients and the medical science filed, Just as ES cell expert Jeanne Loring of Scripps Research Institute said: “both teams should be congratulated and this is a notable moment in the history of human stem cell research”. And he hopes that this will be the beginning of the golden age of stem cell therapeutics.

About us

Creative Bioarray is one of the world’s largest providers of cell services and products. We produces the world's most comprehensive list of research-use cells, including tumor cells, primary cells, stem cells and transformed cells. And we also offer a wide range of cell services related to customized cells, immortalized primary cells, customized iPS cells, stem cell characterization, stem cell differentiation, stem cell assay development/screening, cell culturing sterility testing, toxicology assay and mycoplasma detection & elimination.