Protein arrays enable the analysis of the biological function / activity of dozens to hundreds of proteins in one experiment (e.g. auto-antibody detection, protein-protein interactions, small molecule screening…). In this post, let’s review the recent Raybiotech Protein Arrays focusing on Immunology.
The Picokine™ line of ELISA kits is the culmination of over 20 years of development. Today, I would like to tell you a bit more about this innovation by Boster for your ELISA tests. [Read more…]
All cell types are in contact with the ExtraCellular Matrix (ECM), a complex and dynamic network of macromolecules with different physical and biochemical properties. The ECM plays an essential rule in structural support, adhesion, movement and cell regulation. Its components are frequently monitored, which requires robust and user-friendly kits.
In this post, I invite you to take a look at 5 popular kits you should know about… to boost your ECM studies!
Immune checkpoint molecules play an important role in T cell functionality after TCR/MHC signaling. In fact, blockade of two B7/CD28 family checkpoint molecules, namely CTLA-4 and PD-1, have already demonstrated excellent efficacy in increasing T cell responses to a variety of tumors. [Read more…]
RIPA buffer is one of the most useful protein extraction buffers and maintains most native structures of proteins. Nevertheless, RIPA buffer is not convenient enough to extract membrane proteins and membrane-associated proteins concentrated in lipid raft.
Lipid raft is a highly unique microdomain on the lipid bilayer which contains special lipids, cholesterol and functional proteins. In this layer, lipid raft-enriched proteins are usually insoluble in mild detergent buffers such as 1% Triton X-100 and RIPA buffer. Consequently, it was, up to now, difficult to analyze the functions of this kind of proteins.
We need to find biomarkers for prognostic, diagnostic and personalised treatment development. Notably to fight cancers that affect tissues. Since biopsies are invasive, it’s better to look for biomarkers in body fluids. Indeed, a simple blood sample becomes a kind of ‘liquid biopsy’ to reveal tissues affections. For 13 years, increasing interest has been shown for miRNA as biomarkers and it will last for sure. The 2 main reasons are that they are major regulators of cell processes and they are released from tissues into the blood. They are major biomarker candidates in serum and plasma. Thus, these circulating miRNA (cmiRNA) are the best hope for modern medicine. Still, a lot of research has to be done to determine the specific signature for each pathology, and also depending on the patient background. Obviously, cmiRNA profiling is a key step and requires sensitive and reproducible method. Sequencing, qRT-PCR, several kind of microarrays… Let’s explore together what the best approach could be. [Read more…]
For humans, IFN-α consists of a group of proteins that are greater than 85% homologous by amino acid sequence. A lot of individual human IFN-α subtypes have been identified and many have different properties. For the moment, we don’t know exactly why there are multiple IFN-α subtypes. A variety of studies suggested they possess overlapping but also unique sets of biological activities. Quantification of IFN is essential but usually methods don’t take account all IFN types. In this post, I would like to focus on a relevant ELISA kit to avoid this problem.
Stem Cells: a great tool for biomedical research! From the embryo at a very early stage of development, stem cells have two important capabilities: to multiply to infinity by simple division and to give rise to all kind of cells of the organism. These properties offer many opportunities, not only for the regenerative medicine but also for the study of genetic diseases and development of new treatments.
One of the first thing to do when you’re working on this kind of cells is to check if they are really stem cells, i.e their stemness. It can be highlighted by different markers by IF, WB, etc…
Today, I invite you to look at a popular antibody allowing you to monitor the level of differentiation of your cellular model as well as an innovation related to antibody array and stem cell research.
Early in 2015, researchers of The University of Queensland Diamantina Institute (Australia) have shown a very sensible approach to the discovery of new biomarkers associated to transition from non-metastatic tumours to metastatic tumours in osteosarcoma. Not to be a spoiler, but they found that the uPA/uPAR axis is crucial for this, and can be used as a prognostic biomarker. In fact, inhibition of this axis can inhibit the metastasis in this type of tumours. (Endo-Muñoz et al. DOI: 10.1371/journal.pone.0133592).
I don’t want to focus on the biomarker per se, but rather, on the process that this lab followed to discover this new biomarker. [Read more…]
New techniques such as cDNA microarrays have enabled us to analyse global gene expression. However, almost all cell functions are executed by proteins, which cannot be studied simply through DNA and RNA techniques. In fact, experimental analysis clearly shows disparity can exist between the relative expression levels of mRNA and their corresponding proteins (1).
Therefore, analysis of the proteomic profile is critical, especially in processes that rely on secreted proteins (e.g. inflammation). The conventional approach to analysing multiple protein expression levels has been to use 2-D SDS-PAGE coupled with mass spectrometry. However, these methods are slow, expensive, labor-intensive and require specialised equipment. Moreover, these traditional methods of proteomics are not sensitive enough to detect most secreted biomarkers (typically at pg/ml concentrations).
For some years now, antibody arrays have been available to study markers and publish their discoveries in various areas like Immunology, Atherosclerosis, Inflammation, Angiogenesis, Immunoediting and even signaling pathways (ex. phosphorylation, Receptor Tyrosine Kinases…). So far, however, and in spite of the growing demand by researchers working on stem cells, there were no antibody arrays for this area of research, meaning that individual Western Blots had to be performed. But not any more! [Read more…]