Pathway Focus: Neuroscience/Neuro Stem Cells
- Speed—more rapid results than immunocytochemistry (as little as 15 minutes)
- Multiplexing—simultaneously measure neurite outgrowth and cell viability
- Convenience—use with either live or fixed cells
The dual-color Neurite Outgrowth Staining Kit uses a select combination of fluorescent and background suppression dyes that allows simple, rapid visualization and relative quantification of neurite outgrowth with cell viability.
The ability to analyze and quantify neuronal cell viability is essential in neuroscience. Neurite outgrowth is an important morphological phenotype of neuronal cells that correlates with their function and cell health. As such, it is a key process that can be monitored for assessing the effect of drugs, environmental factors, and biological modifiers on neuronal cells.
The assay protocol involves just a few steps (compared to more laborious immunostaining methods), which allows data collection in as little as 15 minutes.
- Learn more about neurite outgrowth
|Figure 1. Visualization of primary rat cortex neurons using the Neurite Outgrowth Staining Kit. Cryopreserved primary rat cortex neurons were thawed and plated in 96-well format and grown in Neurobasal® medium supplemented with B-27® serum-free supplement for 7 days prior to live-cell staining with the Neurite Outgrowth Staining Kit. Overlay image of the orange-fluorescent Cell Membrane Stain and green-fluorescent Cell Viability Indicator in the same field of view.|
For Research Use or Manufacturing of Cell, Gene, or Tissue-Based Products. CAUTION: Not intended for direct administration into humans or animals.
- Validated—designed for use with the CTS™ line of neural cell culture products
- High performance—performs equivalently to B-27® Serum Free, the industry standard for neural cell culture
- Expanded use—for research use or manufacturing of cell, gene, or tissue-based products
B-27® Supplement XenoFree CTS™ is the complete B-27® Serum-Free Supplement formulated using only recombinant or humanized components.
Designed with the translational stem cell researcher in mind, this 50X B-27® Supplement can be used to support induction of human neural stem cells (hNSCs) from pluripotent stem cells (PSCs), expansion of hNSCs, differentiation of hNSCs, and maintenance of mature differentiated neurons in culture. Additionally, this product is manufactured under cGMP guidelines, contains fully traceable components, and raw materials have been tested for adventious agents.
Researchers interested in transitioning their work from the bench to the clinic have a vested interest in using reagents that are manufactured in controlled conditions (cGMP), that contain xeno-free, traceable raw materials, and that have been tested for adventitious agents. Through its unique formulation, rigorous quality testing, and availability of certificates of origin, B-27® Supplement XenoFree CTS™ offers an advantage to researchers submitting IND applications.
Learn more about the Gibco® CTS™ line of neural cell culture products
Figure 2. iPSC differentiation. iPSC were differentiated in xeno-free neural induction medium made with B-27® Supplement XenoFree CTS™ supplements and small molecules. Derived NSC (A) were characterized with phenotype marker with Sox1 (B) and Nestin (C). The population was devoid of cells with pluripotent marker of Oct4 (D). Upon further differentiation, neurons (E, DcX, red) and oligodendrocytes (F, GalC, green) were obtained. All nuclei were stained with DAPI (blue).
For Research Use Only. Not for use in diagnostic procedures.
- ABfinity™ rabbit recombinant antibodies are screened to be sensitive and specific
- Enable consistent performance for your assays
- Consistent product from lot-to-lot
ABfinity™ recombinant rabbit antibodies are produced from specific recombinant clones, so antibody performance is consistent over time. Lot-to-lot variability, commonly a problem with polyclonal products, is minimized with ABfinity™ antibodies. In addition to the monoclonal versions of our ABfinity™ antibodies, we offer oligoclonal versions of several of these specificities. Oligoclonal ABfinity™ products are a mixture of recombinant monoclonal antibodies and offer the improved signal strength you get with polyclonal antibodies, combined with the highly reproducible results you get from monoclonals. ABfinity™ antibodies are available to many targets including beta endorphin (Gene ID 5443).
Beta endorphin is an opioid neuropeptide that is produced either by neurons of the pituitary gland and released into the circulatory system, or by the hypothalamus and released into the brain and spinal cord through nerve fibers. The precursor polypeptide for beta endorphin is pro-opiomelanocortin (POMC), which can also be cleaved to produce adrenocorticotrophic hormone (ACTH).
- Learn more about ABfinity™ Recombinant Rabbit Monoclonal Antibodies
- Use the Primary Antibody Search Tool
|Figure 3. Western blot analysis of recombinant beta endorphin (40 ng). Beta endorphin was detected at ~ 3 kDa using ABfinity™ Beta Endorphin Recombinant Rabbit Oligoclonal Antibody at a concentration of 2 μg/mL. The blot was developed using enhanced chemiluminescence (ECL) method.|
For Research Use Only. Not for use in diagnostic procedures.
- Sensitive—high resolution and image contrast allow visualization of fine neuronal
- Precise—multicolor labeling allows simultaneous identification of multiple origins and targets
- Powerful—histochemical processing required for image development
Tested and validated in thousands of research laboratories worldwide, we offer a broad range of reagents for neuronal analysis: anteretrograde and retrograde tracing, measurements of gap-junction communication, anatomical targets (e.g. Nissl bodies and myelin), and physical form (solid, liquid, crystal). Ultrabright and photostable Alexa Fluor® conjugates of dextran, cholera toxin, and WGA help ensure that your images will accurately reproduce biological structures. Our proprietary solutions for histological staining of myelin (FluoroMyelin™ kits) and Nissl staining (NeuroTrace® stains) significantly speed your time to results and help minimize the unpredictable outcomes associated with traditional approaches.
Fluorescent and biotinylated tracers can be used to define neuronal cell connectivity, to study dye translocation through gap junctions, and to mark or identify specific cells of interest. Life Technologies offers a comprehensive set of reagents for neuronal tracing and anatomy.
The sensitivity and dynamic range of fluorescence is ideal for visualization of neuronal network structure, function, and connectivity. The high signal-to-noise is particularly well suited for tissue analyses where cellular complexity and depth pose challenges for other methods. Some fluorophores can also be used as haptens with anti-dye antibodies for signal amplification of weak signals (using techniques such as TSA™ (Tyramide Signal Amplification) or the photo-conversion of diaminobenzidine (DAB)). Biotinylated tracers, on the other hand, require detection with a fluorescent dye– or enzyme-labeled conjugate of streptavidin.
|Figure 4. Pyramidal cells of the hippocampus and dentate gyrus in a transverse cryosection of paraformaldehyde-fixed mouse brain. NeuroTrace® green fluorescent Nissl stain is localized to neuronal somata, while non-neuronal cells can be identified by the presence of DAPI-stained nuclei. This image is a composite of images taken using a 10x objective and filters appropriate for fluorescein and DAPI.|
Fluorescent Histological Stains
Protein Conjugates for Neuronal Tracing
TSA Signal Amplification
- Fast and efficient labeling
- Sensitive viability indicator
- Extremely bright morphological marker
Non-fluorescent calcein AM easily permeates intact, live cells. Hydrolysis of calcein AM by intracellular esterases produces calcein, a strongly fluorescent compound that is well-retained in the cell cytoplasm. The bright fluorescence permeates the neuronal body and dendrites, providing an excellent morphological marker. As dead cells and cells with compromised cell membranes do not retain calcein and have lower esterase activity, calcein AM provides a simple, rapid, and accurate method to measure cell viability and cytotoxicity.
The easy protocol and extremely bright signal makes calcein AM ideal for visualization of neuronal bodies and dendrites for morphological and functional studies of neurons in culture, for instance in measuring neurite volumes and outgrowth. In quantitative measurements the calcein signal is a highly sensitive and accurate measure of the number of healthy, living cells and, conversely, of neuronal damage, death, and toxicity.
Calcein AM is an ideal reagent for studies of neuronal differentiation, proliferation, and health. The exceptionally bright staining provides assay accuracy and robustness.
|FIgure 5. NG108 cells (mouse neuroblastoma x rat glioma hybrid) were loaded with calcein AM and counterstained with NucBlue™ Live Cell Stain.|
- Room temperature–stable formulation of high-purity Hoechst 33342 and DAPI
- Ready-to-use with no need to dilute, weigh, or pipette
- Nuclear staining is achieved with just two drops per milliliter of solution
Fluorescence imaging is often challenging and tedious, with more time spent preparing samples and reagents than acquiring images and analyzing results. To help researchers focus more on biology and get results faster, scientists at Life Technologies applied their experience in Molecular Probes® fluorescence technologies and cell-based applications to develop a new family of imaging reagents that emphasizes user friendliness and work flow convenience—ReadyProbes™ Ready-to-Use Reagents.
NucBlue™ Live Cell and Fixed Cell Stains are some of our most popular ReadyProbes™ reagents. Combining the classic and popular nuclear stains Hoechst 33342 and DAPI, NucBlue™ stains are room temperature–stable solutions provided in dropper bottles for your convenience. Two drops per milliliter of solution is all you need.
Our ReadyProbes™ reagents—including NucBlue™ Live and Fixed Cell nuclear stains—add convenience and reliability to your imaging experiments.
- Learn about ReadyProbes™ Ready-to-Use Imaging Reagents
|Figure 6. NG108 cells (mouse neuroblastoma x rat glioma hybrid) were labeled with NucBlue™ Live Cell Stain and imaged on a Nikon confocal microscope.|
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