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Label IT siRNA Tracker Intracellular Localization Kit, Cy3 – labels 50 µg siRNA

Efficient, direct labeling for in vitro and in vivo tracking of siRNA

SKU MIR 7212 Categories ,

$519.00

Description

  • 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

 

Kit Components:
Each Kit contains Label IT® siRNA Tracker™ Reagent, Label IT® Reconstitution Solution, 10X Labeling Buffer A, and siRNA Dilution Buffer.

 

 

SKU: MIR 7212

Supporting Data

 

 

LabelIT General LabelIt Schematic Data Illustration

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.

HeLa cells in 12-well plates were transfected at 70% confluence with TransIT-TKO® Transfection Reagent (3 μl/well) and Label IT® siRNA Tracker™ Fluorescein-labeled siRNA duplexes (GREEN, 50 nM final concentration in the well)

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 Label IT® siRNA Tracker™ Fluorescein-labeled siRNA duplexes (GREEN, 50 nM final concentration in the well). 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 siRNA.

Delivery of Labeled siRNA Data

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.

Resources

Specifications

Storage Conditions
All Configurations: Store the Label IT® Reagent at -20°C in both its dried pellet and reconstituted form. Store all other supplied reagents at -20°C. Warm kit components to room temperature and quick spin before each use.

Product Guarantee
All Configurations: 1 year (6 months after reconstitution)

Usage Statement
All Configurations: For Research Use Only.

Animal Origin Statement
All Configurations: This product is animal origin free.

 

Technical Product Literature

Full Protocol
Label IT siRNA Tracker™ Kit Full Labeling Protocol (PDF)

Quick Reference Protocol
Label IT siRNA Tracker™ Kit Quick Ref Protocol (PDF)

SDS
Label IT siRNA Tracker™ Kit SDS (PDF)

 

Additional Resources

Technical Report: High Efficiency siRNA Delivery In Vitro (PDF)
Technical Report: Delivery Tools for Neurobiology (PDF)
For more information on siRNA delivery in vivo, see the following references:
Lewis et al.(2002) Nature Genetics 32: 107-107.
Zhang et al.(1999) Human Gene Therapy 10: 1735-1737.
Liu et al.(1999) Gene Therapy 6: 1258-1266.
Hagstrom et al.(2004) Molecular Therapy 10: 386-398.

FAQs

See the Label IT FAQs

Using transfection reagents and enhancers from Mirus has given our platform a competitive advantage. Increasing efficiency and lowering costs for all of our porgrams.

 

Sally Mader, PhD
ZYZ Theraputics