Label IT® siRNA Tracker Intracellular Localization Kits
Efficient, direct labeling for in vitro and in vivo tracking of siRNA
The Label IT® siRNA Tracker Kits.
The Label IT® siRNA Tracker labeling reagents are composed of three regions: the label (a fluorophore or biotin) (green), the linker (yellow) which facilitates electrostatic interactions with the siRNA and miRNA and the reactive alkylating group (blue) that covalently attaches the Label IT® reagents to any reactive heteroatom within the siRNA or miRNA. Attachment of the Label IT® Reagent to siRNA or miRNA is optimized and does not alter their structure or affect downstream target knockdown performance. The labeled siRNAs or miRNAs can be both visualized after transfection by fluorescence microscopy and simultaneously employed to knockdown target gene expression.
Successful Delivery and Functional Knockdown using Labeled siRNA.
(A) HeLa cells in 12-well plates were transfected at 70% confluence with TransIT-TKO® Transfection Reagent (3 µl/well) and 50 nm/well of Label IT® siRNA Tracker Fluorescein-labeled siRNA duplexes (GREEN). The cells were incubated 24 hours post-transfection then fixed and counterstained with TO-PRO®-3 (nuclei, BLUE) and Alexa Fluor® 546 Phalloidin (actin, RED). Confocal images were acquired on a Zeiss LSM 510 Confocal Microscope. (B) TransIT-TKO® Transfection Reagent was used to transfect anti-firefly luciferase siRNA into CHO-Luc cells stably expressing firefly luciferase. The siRNA was either unlabeled or labeled with Label IT® siRNA Tracker Cy®3, Cy®5, Fluorescein, or CX-Rhodamine Reagents. Bars indicate the percent firefly luciferase expression 24 hours after delivery of 5 nM anti-firefly luciferase mRNA.
Delivery of Labeled siRNA to Mouse Liver.
(A) siRNA (25 µg) was labeled with Label IT® siRNA Tracker Cy®3 Kit and delivered to mice through the tail vein using either low volume injection conditions (100 µl) hydrodynamic injection conditions (2 ml over 6-8 seconds). (B) Mouse livers were harvested 30 minutes after injection, fixed in paraformaldehyde, sectioned (10 µm thick) and analyzed. A representative hepatocyte (h) and sinusoid (s) are indicated in each panel. The hydrodynamic injection of siRNA results in siRNA uptake by the majority of the hepatocytes in the liver.
- Versatile Labeling – Efficiently label and visualize the siRNA of your choice.
- One-step Chemical Method – Easily and consistently control the labeling reactions and labeling density.
- High Efficiency Labeling – Optimal visualization of siRNA in cells and tissues.
- The Label IT® siRNA Tracker Intracellular Localization Kit provides a straight-forward approach to directly label siRNA of any sequence containing guanine residues, in an efficient and non-destructive manner for tracking experiments. This kit allows tracking of functional fluorescently labeled siRNA in mammalian cells and enhances the ability to visually assess siRNA transfection efficiency. Cellular uptake, subcellular localization, and functional inhibition of target gene expression can be monitored in the same experiment following the introduction of the labeled siRNA into mammalian cells. This kit provides sufficient reagents to label 50 µg (5 x 10 µg reactions) of siRNA.
Fluorophore Spectral Characteristics
|Fluorophore||Excitation Wevelength or λmax(nm)||Emission Wavelength (nm)|
- Each Kit contains Label IT® siRNA Tracker Reagent, Label IT® Reconstitution Solution, 10X Labeling Buffer A, and siRNA Dilution Buffer
- For shipping and storage information please click on Order#.
|MIR7212||Label IT® siRNA Tracker Intracellular Localization Kit, Cy®3||labels 50 µg siRNA||421,00|
|MIR7213||Label IT® siRNA Tracker Intracellular Localization Kit, Cy®5||labels 50 µg siRNA||421,00|
|MIR7214||Label IT® siRNA Tracker Intracellular Localization Kit, CX-Rhodamine||labels 50 µg siRNA||385,00|
|MIR7215||Label IT® siRNA Tracker Intracellular Localization Kit, TM-Rhodamine||labels 50 µg siRNA||385,00|
|MIR7216||Label IT® siRNA Tracker Intracellular Localization Kit, Fluorescein||labels 50 µg siRNA||385,00|
|MIR7217||Label IT® siRNA Tracker Intracellular Localization Kit, Biotin Labeling||labels 50 µg siRNA||385,00|
- All prices are in EURO excl. VAT and shipping. Only available in selected countries.
- Hagstrom JE1, Hegge J, Zhang G, Noble M, Budker V, Lewis DL, Herweijer H, Wolff JA: A facile nonviral method for delivering genes and siRNAs to skeletal muscle of mammalian limbs. Mol Ther. 2004 Aug;10(2):386-98. Pubmed
- Lewis DL, Hagstrom JE, Loomis AG, Wolff JA, Herweijer H: Efficient delivery of siRNA for inhibition of gene expression in postnatal mice. Nature Genetics 32: 107-8, 2002. Pubmed
- Zhang G, Budker V, Wolff JA: High levels of foreign gene expression in hepatocytes after tail vein injections of naked plasmid DNA.Hum Gene Ther. 1999 Jul 1;10(10):1735-7. Pubmed
- Liu F, Song Y, Liu D: Hydrodynamics-based transfection in animals by systemic administration of plasmid DNA. Gene Ther. 1999 Jul;6(7):1258-66. Pubmed