Read the latest issue of the BioPath Online E-Newsletter.
Pathway Focus: Neurodegeneration
- Visualize LC3B recruitment to the autophagosome
- Deliver GFP- or RFP-tagged LC3B to cells using reliable BacMam 2.0–based kits
- Choose the LC3B Antibody Kit for fluorescence microscopy and high-content imaging and analysis
Autophagy is a cellular process involved in degradation of organelles and proteins and is important for neural homeostasis, with dysfunction linked to a number of disease conditions. One of the most common methods used to study autophagy is the visualization of the recruitment of LC3B protein (Entrez gene ID: 81631; aliases: MAP1A/1BLC3) to autophagosomes.
The Premo™ Autophagy Sensor combines the selectivity of an LC3B-GFP or LC3B-RFP chimera with the transduction efficiency of BacMam technology, enabling unambiguous visualization of this protein in live cells. The sensor reagent also tolerates fixation with formaldehyde and thus is compatible with fixed-cell analysis.
The LC3 Antibody Kit for Autophagy includes an anti-LC3B rabbit polyclonal antibody that has been validated for use in fluorescence microscopy and high-content imaging and analysis. This kit also includes a control compound for inducing autophagosomes.
These tools allow the highly dynamic processes occurring during autophagy, from autophagosome formation to degradation in the autolysosome, to be studied in a relevant cellular setting.
Learn More About Autophagy Sensors
|Figure 1. Imaging primary cells with CellLight® reagents. Primary human skeletal muscle cells (GIBCO® HSkM-S cells) were cultured in poly-Dlysine–coated glass-bottomed petri dishes and transduced with CellLight® ER-GFP and CellLight® Golgi-RFP in complete medium. The following day, the cells were loaded with 1 μg/mL Hoechst 33342 and 25 nM MitoTracker® Deep Red stains for 5 min at 37°C in DPBS. Cells were washed three times andimaged using standard DAPI/FITC/TRITC/Cy®5 filters.|
- ABfinity™ phosphospecific antibodies for LRRK2, tau, and AKT: rabbit recombinant antibodies for sensitive detection
- 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. ABfinity™ antibodies are available against many targets, including LRRK2, tau, and AKT (gene IDs: 120892, 4137, 11651).
The search for therapeutic targets in Parkinson’s disease and Alzheimer’s disease has implicated different kinases, from mitogen-activated protein kinase (MAPK) family members to new mutants of mixed lineage kinase (MLK)-like kinase and leucine-rich repeat kinase 2 (LRRK2), as potential targets in regulating dopamine neuron viability. In some treatments, AKT has been demonstrated to mediate important neurotrophic and anti-apoptotic effects. As research leads the investigation of these neurodegenerative diseases through various signaling cascades, antibodies to kinases and downstream targets will help assess the expression patterns of these potential components in neurodegenerative diseases in model systems.
Learn more about ABfinity™ Recombinant Rabbit Monoclonal Antibodies
Use the Primary Antibody Search Tool
|Figure 2. Immunocytochemistry analysis of HeLa cells with AKT [pT308] Rabbit Recombinant Oligoclonal Antibody. Alexa Fluor® 488 goat anti-rabbit IgG was used as a secondary antibody (green), DAPI stains the nucleus (blue), and Alexa Fluor® 594 phalloidin stains actin (red). (A), (B) Composite images of cells showing nuclear localization of phosphorylated AKT. (C) Competition with the phospho-AKT [pT308] peptide.|
Gibco® Recombinant PDGF products offer:
- High biological activity
- High purity
- Proven compatibility with Gibco® media
Gibco® recombinant neurotrophic growth factors are highly purified, bioactive proteins that can be used in combination with Gibco® specialty media products to stimulate growth of various neuronal cells for use in your research.
Gibco® cytokines and growth factors are high-purity recombinant proteins with high bioactivity. So that Gibco® growth factors are of the highest quality, each protein is analyzed for purity along with structural homogeneity to help ensure a biologically active protein. And Gibco® cytokines and growth factors have been bioassayed with Gibco® media. Get your cytokines and growth factors from the media company you use and trust.
Learn more about Gibco® Cytokines and Growth Factors
- Enables reliable, consistent measurement of tau proteins
- Rapid results—4-hour incubation protocols
- Flexible—precoated, breakable strip-well plates
Tau is a highly soluble microtubule-associated protein, expressed in neurons and the central nervous system, that promotes microtubule assembly and stability. Posttranslational modifications modulate tau’s activity—phosphorylation decreases the ability of Tau to bind to microtubules, and hyperphosphorylation can result in the self-assembly of tangles of paired helical filaments and straight filaments, which are involved in the pathogenesis of Alzheimer’s disease and other tauopathies.
Tau is phosphorylated at multiple sites by a variety of kinases, such as GSK3, PKA, CK-1, CDK1, CDK5, MAPK, and MARK. Glycogen synthase kinase-3β is a critical modulator of cell fate, neuronal plasticity, and tumorigenesis. It phosphorylates tau at multiple sites, including serine residues 199 and 396. It is hypothesized that a priming phosphorylation by kinases such as PKA or CK-1 greatly accelerates the actual phosphorylation of threonine residue 231. Of the multiple potential phosphorylation sites of tau, threonine 181 is important as well.
We have developed assay kits to monitor site-specific phosphorylation events using ELISA. These assays have been extensively characterized to show linear responses to natural samples, specificity to a given phosphorylation site, and high sensitivity and precision for accurate measurement of tau phosphorylation. Total tau protein can be measured using the total tau ELISA kits.
Learn more about ELISA Kits
|Figure 3. The Tau [pT231] Human ELISA Kit is specific for the measurement of tau [pT231]. To determine the specificity of this ELISA kit, cell extracts from different cell lines, each at a concentration of 200 µg/mL total protein, were analyzed. The figure shows that the kit detects Tau [pT231] in cell lysates from SH-SY5Y and MCF-7 human cells.|
|After more than 30 years in print, the award-winning BioProbes® journal for cell biology applications is going green in a new electronic format, dramatically reducing the amount of natural resources that are typically consumed to produce and transport paper copies. Highlighting the latest and greatest Molecular Probes®, Gibco®, Dynal®, and Invitrogen™ products for cell biology applications, BioProbes has been a trusted source for many readers. As a subscriber to BioPath Online, we thought you might be interested in BioProbes. Follow the link below to register your email address, and you’ll receive a notification each time a new issue is published.
Learn more about BioProbes Going Green