Application Research of ER-Tracker Red Fluorescent Probe for Endoplasmic Reticulum Imaging in Live Cells
1. Basic Physicochemical Properties of ER-Tracker Red
ER-Tracker Red is a red fluorescent probe specifically designed for endoplasmic reticulum imaging in live cells, with the molecular formula C44H42BClF2N6O7S2 and a molecular weight of 915.2316. This probe has optimal excitation efficiency at an excitation wavelength of 587 nm, with an emission peak at 615 nm, perfectly matching the TRITC filter system used in conventional microscopy.
In terms of physical state, ER-Tracker Red is provided in liquid form with a purity guaranteed to be over 95%, ensuring the reliability and reproducibility of experimental results. To ensure probe stability, it is recommended to store it at -20°C in a light-protected dry environment to avoid repeated freeze-thaw cycles. It should be noted that this probe solution can be stored short-term under conditions of 4°C; however, long-term storage must adhere to the standard condition of -20°C.
2. Structural Characteristics and Mechanism of Action of ER-Tracker Red
The molecular design of ER-Tracker Red is highly distinctive; its core structure consists of two parts: one as the fluorescent reporting group (BODIPY TR fluorescent dye) and the other as the targeting group (Glibenclamide). This clever molecular design provides excellent fluorescence performance along with precise organelle localization capabilities.
Its mechanism primarily relies on Glibenclamide's specific binding to ATP-sensitive potassium ion channel receptors (KATP channels) on endoplasmic reticulum membranes. This binding exhibits high selectivity, allowing the probe to accurately localize within endoplasmic reticulum membrane structures without significantly labeling other organelles. Compared to traditional non-specific dyes like DiOC6(3), ER-Tracker Red improves labeling efficiency for endoplasmic reticulum by three to five times while reducing non-specific staining for mitochondria by over 80%.
3. Main Application Advantages of ER-Tracker Red
3.1 High Specificity for Endoplasmic Reticulum Labeling ER-Tracker Red’s most notable advantage lies in its high specificity for labeling endoplasmic reticulae. Experimental data show that at a working concentration of 1μM, this probe can clearly delineate endoplasmic reticular structures while almost completely ignoring non-specific staining from other membrane structures such as Golgi apparatus or lysosomes—making it an ideal tool for studying morphological and dynamic changes within endoplasmic reticulae. 3.2 Excellent Cell Compatibility Regarding cytotoxicity, ER-Tracker Red performs exceptionally well; even during continuous observation lasting up to 72 hours using concentrations ranging from 1–5μM shows no significant impact on key indicators such as cell proliferation vitality or apoptosis rates compared with approximately a30% decrease seen when using traditional dye DiOC6(3) under similar conditions—this low toxicity characteristic makes it particularly suitable for prolonged live-cell dynamic observation experiments. 3.3 Multifunctional Research Platform The application range covered by ER-Tracker red is extensive; In pancreatic β-cell studies—it can monitor real-time dynamics occurring within insulin secretion processes related directly back towards their respective intracellular environments where they originate—from cardiac myocyte investigations observing stress-induced responses occurring throughout these same regions! More importantly still—is how this particular reagent pairs effectively alongside others like mitochondrial probes (i.e., Mito Tracker)—allowing researchers insight into interactions between various cellular compartments!
Four: Comparison With Related Reagents
influencing Cellular Organelles' Fluorescence Markers There are numerous functional analogs available today aimed explicitly toward investigating mitochondria-related phenomena including but not limited too MitoPerox & Mito Tracker RED CMXRos whilst classic ROS fluorescence markers include CM-H2DCFDA plus calcium ion detection methods utilize either Fluo5N AM(cross-membrane permeable) OR Fluo8 sodium salt(non-permeable). nIt’s worth noting each marker possesses unique characteristics requiring researchers carefully consider selecting appropriate tools based upon individual experimental needs—for instance combining both ERT tracker AND mito tracker green would yield desirable dual-color marking effects when examining contact points between ENDOPLASMIC RETICULUM-MITOCHONDRIA interfaces known colloquially referred-to-as ‘MAMs’! n### Five: Usage Precautions As professional research reagents only intended laboratory use strictly prohibited food/human applications!! During experimentation care taken maintain light protection conditions suggested utilizing CO₂ incubators uphold physiological standards during staining procedures varying types cell lines require preliminary gradient testing determine optimal working concentrations typically starting test ranges set between .05-.05mM!. Staining duration adjustable according experiment requirements generally falls anywhere around fifteen thirty minutes post-staining washing pre-warmed media/buffer solutions advised removing unbound molecules thereafter fresh media replacement every hour/two maintaining normal physiology states.