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Introduction Transformation of bacteria with plasmids is not only of note for bacterial studies but also can be used for gene expression studies in mammalian cells. Most plasmids are of bacterial origin and contain both a bacterial origin of replication and an antibiotic resistance gene which can be used as a selectable marker. The process of transformation allows foreign DNA to be introduced into a cell. Genetic modifications can be made to various bacterial strains to make them more amenable to transformation. Such modifications will maintain the plasmid without rearranging the plasmid DNA. Certain treatments have been shown to increase the transformation efficiency of bacteria. …

Radial glial cells are essential in shaping the nervous system, serving dual roles in neurogenesis and structural support during the brain's developmental stages. Key Takeaways: Radial glial cells are vital in neurogenesis and brain development, acting as both progenitor cells and structural guides. They exhibit unique radial morphology, extending from the ventricular zone to the pial surface. Radial glial cells differentiate into various neuronal and glial types, influencing neural circuit formation. Table of Contents Jump to a section: - Cell Identification - Radial Glial Stem Cells - Glial Cell Development - Functions and Impacts of RGCs - T …

Cell Signalling · Growth FactorsGrowth Factors: Families, Receptors and Signalling PathwaysGrowth factors are secreted proteins that instruct cells whether to divide, differentiate, migrate or survive. Despite covering families as different as FGF, TGF-beta, EGF, insulin, VEGF and PDGF, they converge on a small number of receptor types and an even smaller number of downstream pathways. This guide covers each major family, the receptor class it uses, and why that distinction matters more than the ligand itself.Browse growth factor ELISA kits →8Major families covered23Members of the FGF familyRTKsThe dominant receptor classPI3K & MAPKThe shared downstream pathwaysKey takeawaysGrowth factor …

Gastroenterology · Barrier BiologyZonulin and Intestinal PermeabilityZonulin is the best-characterised endogenous regulator of intestinal tight junctions — the protein that opens the gut barrier on demand, and the one most often invoked to explain what is popularly called leaky gut. This guide covers what zonulin is, how it loosens tight junctions, which conditions it has been linked to, and how it is measured in the laboratory.Browse barrier assays →pre-HP2The protein identified as zonulin16Chromosome carrying the HP gene4Families of tight junction protein2Best-established triggers: gluten and bacteriaKey takeawaysZonulin is a signalling protein that reversibly loosens the tight junctions b …

Introduction Cellular senescence is a fascinating phenomenon that plays a crucial role in the aging process and various age-related diseases. It is essential to explore the cellular sources, how cells age, and define the concept of senescence to gain a comprehensive understanding of this complex biological process. Key Takeaways Cellular senescence is a state of irreversible growth arrest with significant implications for aging and age-related diseases. Strategies to counteract senescence include senolytics, targeting key pathways, and lifestyle interventions. Senescent cells contribute to tissue dysfunction and chronic inflammation through the senescence- …

Oct-4 stem cells, also known as Octamer-binding transcription factor 4 stem cells, are a fascinating area of study in the field of stem cell research. These stem cells hold great promise due to their unique characteristics and potential applications in regenerative medicine and developmental biology. This article provides information on Oct-4 stem cells, their properties, roles, and therapeutic implications. What are Oct-4 Stem Cells? Octamer-binding transcription factor 4 stem cells (Oct-4) stem cells are a subtype of pluripotent stem cells that express the Oct-4 transcription factor. Belonging to the POU family of homeodomain transcription factors, Oct-4 …

Stem Cells: What are they and how they can help treat diseases Stem cells are a unique population of cells that possess the ability to self-renew and differentiate into specialized cell types. They play crucial roles in tissue maintenance, repair, and regeneration throughout life. Stem cells have the potential to treat diseases, injuries, and degenerative conditions. Stem cells are used medical procedures like bone marrow transplants. Stem cells' versatile nature and vital functions make them a subject of intense scientific exploration and therapeutic potential. { "@context": "https://schema.org", "@type": "BlogPosting", "mainEntityOfPage": { "@type": "WebPa …

Biochemistry · Cell SignallingPhosphorylation: Types, ATP Synthesis and Cell SignallingAdding a phosphate group is how cells both store energy and switch proteins on and off. The same chemistry underlies ATP synthesis in mitochondria and chloroplasts and the regulatory phosphorylation that drives every signalling cascade. This guide covers the three metabolic routes to ATP, how protein phosphorylation works as a switch, and how phosphorylation is measured.Browse phospho antibodies →3Routes to ATP: substrate-level, oxidative, photo~30–32ATP per glucose, current estimateSer / Thr / TyrResidues phosphorylated in proteinsATP synthaseCouples proton gradient to synthesisKey takeawaysPhosphorylatio …

Cell Biology · PathologyNecrosis: Types, Causes, Mechanisms and How It Is MeasuredNecrosis is cell death accompanied by membrane rupture and release of intracellular contents — which is what makes it inflammatory, and what distinguishes it from apoptosis. It was long regarded as purely uncontrolled, but a large part of it is now known to be actively regulated. This guide covers the classical morphological patterns, the causes, the regulated pathways, and the assays used to detect and distinguish them.Browse cell death assays →6Classical morphological patternsLDHStandard membrane-integrity readoutRIPK3 / MLKLNecroptosis effector kinasesHMGB1Archetypal DAMP released on ruptureKey takeawaysNecr …

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Unravel the complexities of bone resorption, a crucial biological process essential for skeletal health and calcium regulation, through the intricate roles of osteoclasts and osteoblasts. Key Takeaways Bone resorption is essential for maintaining skeletal health and calcium balance. Osteoclasts dissolve bone tissue, and osteoblasts create new bone, forming a continuous cycle called bone remodeling. Imbalance in bone resorption can lead to conditions like osteoporosis, hyperparathyroidism, or osteopetrosis. Subperiosteal resorption can indicate systemic diseases, particularly hyperparathyroidism. Calcium, stored in bone as hydroxyapatite crystals, plays a vital role in bone res …

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Lab Methods · Flow CytometryFlow Cytometry Protocol: Two Worked Protocols, Controls and 10 Practical TipsFlow cytometry measures individual cells in a fluid stream at hundreds per second, reporting size, granularity and fluorescence for each one. Most failures come not from the instrument but from sample preparation, panel design and controls. This guide gives two complete worked protocols, a full controls reference table, and ten practical points that determine whether the data is interpretable.Browse flow cytometry reagents →FSC / SSCSize and granularityPMTPhotomultiplier voltageFMOThe control that sets gates9Controls in the reference tableKey takeawaysForward scatter correlates with parti …

Flow Cytometry · Cell SortingGuide to Fluorescence-Activated Cell Sorting (FACS)Fluorescence-activated cell sorting (FACS) is a specialised form of flow cytometry that physically isolates individual cells from a mixed sample based on their fluorescence and light-scattering properties. With sorting purities approaching 99.99%, it is one of the most powerful tools in modern cell biology, immunology and stem cell research.Explore flow cytometry antibodies →99.99%achievable sort purity1000sof cells sorted per second4+parameters measured at onceSinglecell resolutionKey TakeawaysFACS isolates individual cells with a purity approaching 99.99%.It sorts cells by size, granularity, surface markers and …

Cell Signalling · EnzymologyProtein Kinases: Function, Classification, Structure and InhibitorsProtein kinases transfer a phosphate group from ATP onto specific residues of target proteins, switching their activity on or off. Around 500 of them make up the human kinome, and because they sit at the control points of nearly every pathway, they are among the most heavily pursued drug targets in medicine. This guide covers what they do, how they are classified, their conserved architecture, how they are regulated, and how inhibitors work.Browse kinase proteins →~500Protein kinases in the human kinome3Classes by target residueSer / Thr / TyrResidues that get phosphorylatedMg²⁺Cofactor required fo …

Spindle assembly checkpoint The spindle assembly checkpoint (SAC), or mitotic checkpoint, is the main cell cycle control mechanism that governs mitosis. Incorrect spindle attachment causes the spindle assembly checkpoint to generate a wait anaphase signal that prevents the activation of the anaphase promoting complex (APC), an ubiquitin E3 ligase essential for progression of the cell cycle. Conserved components of the mammalian SAC include Bub1 (Cahill et al., 1998), BubR1 [Chan et al., 1998], Bub3 [Taylor et al.,ÿ1998], Mad1[Jin et al., 1998], Mad2 [Li and Benezra, 1996] and CENP-E [Weaver etÿal., 2003]. The checkpoint becomes activated as a …

What are Stem Cells? Stem cells are primitive, undifferentiated, and unspecialised cells that are found in multicellular organisms. Stem cells can be thought of as "uncommitted cells", as they have not yet assumed a specific function in the body. Embryonic stem (ES) cells are present everywhere during early development before the formation of organs and organ systems. Stem cells are stimulated by their environment into becoming specialized cells, such as muscle cells or brain cells. On top of being able to differentiate into the different types of cells and tissues that make up the human body, stem cells maintain their ab …

by Ryan Wallis, PhD Aging and Disease Imagine a world where everything you do is missing just a little piece and as time goes on you start to lose sense entirely. This is the devastating world of neurodegeneration, a collective term for diseases which include Alzheimer’s and Parkinson’s. Conditions like these have an enormous impact on both the individuals who suffer from them and their families, who must watch their loved ones ebb away into a shell of their former selves. These conditions are as cruel as they are debilitating and, alarmingly, they are on the rise (Alzheimer’s Association, 2017). In fact, almost all conditions associated with ageing ar …

Key Points Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR) is a method of gene editing that uses the Cas9 protein and specific guide RNAs to either delete host genes or insert specific gene sequences into the host genome. CRISPR screening is used to find a small number of important genes or genetic sequences within a massive number of genetic sequences such as the entire genome. There are three main types of CRISPR screening assays which include viability-based negative selection screens, viability-based positive selection screens and marker-based selection screens …

RIPA (Radio Immuno Precipitation Assay) buffer is mostly used when carrying out a western blot or immunoprecipatation assay. A RIPA buffer is used in order to lyse cells and extract protein from cultured cells. RIPA buffer cell lysis enables determination of protein concentration. RIPA buffer is an ideal cell lysis reagent since it contains three non-ionic and ionic detergents. Preserving protein:protein interactions When you need to preserve protein-protein interactions or to reduce denaturation its recommended to use a RIPA buffer recipe without SDS (ionic detergent) or Triton X-100 (non-ionic detergent). RIPA buffer recipes …

Bone Morphogenetic Proteins (BMP) were first discovered in the 1960s by Dr.Marshall Urist, an orthopaedic surgeon at UCLA (Urist 1965). BMPs are classically associated with their roles in limb development, induction of cartilage and bone growth. However, it has been since clarified that Bone Morphogenetic proteins (BMPs) are involved in many more diverse biological processes, such as stem cell and organ formation, muscle development, iron metabolism, vascular biology and cancer (D. P. Brazil et al.2015). The BMPs belong to the TGF-β superfamily and are glycosylated, extracellular matrix-associated molecules. The constituents of the BMP signalling pathway have also been implicated in d …

David Melamed, Phd Candidate, Colombia University When I started in the Kalderon Lab five years ago, I had no idea that our work would lead us to drastically revise an entire model system of adult stem cell behavior. I joined the lab straight out of college, setting forth with a wide-eyed passion for stem cell research. The lab was a perfect fit, specializing in the behavior of somatic stem cells found in the fruit fly ovary. Our cells of interest, called Follicle Stem Cells (FSCs), closely mimic the behavior of various somatic adult stem cells in mammals, including those found in the stomach and intestine. Follicle Stem C …

By John Bannon PhD Early Cytokinesis Early cytokinesis; from anaphase to midbody formation cytokinesis is the physical separation of a dividing cell into two identical daughter cells [Eggert et al., 2006]. Cytokinesis begins following the separation of DNA during anaphase, and the consequent APC-mediated destruction of the Cdk1-cyclin B complex, which negatively regulates many proteins essential for cytokinesis [Hummer and Mayer, 2009; Jiang et al., 1998; Mishima et al., 2004]. Microtubule Alignment During early anaphase microtubules are rearranged to form the central spi …

Polo-like kinase (Plk1) Polo-like kinase (Plk1), is a serine/threonine kinase that is active during mitosis (Hamanaka et al., 1995). Its name is derived from its original characterisation as a homolog of a Drosophila mitotic kinase encoded by the Polo gene (Fenton et al., 1993; Martin et al., 2006). There are five Plk kinases identified in humans, although only Plk1 has been extensively characterised as a mitotic kinase (Martin et al., 2006; Lens et al., 2010). Polo-like Kinase family members Other members of the family have largely non-mitotic functions: Plk2 and Plk4 are involved in centriole duplication (Warnke et al., 2004; …