Human ON-TARGETplus RTF - Tyrosine Kinases

The only ready-to-use siRNA screening libraries available


Ideal for labs who want to carry out a few screens but lack high-throughput capabilities.

Human ON-TARGETplus RTF Tyrosine Kinase siRNA Library includes siRNA reagents targeting tyrosine kinases; a large family of evolutionarily conserved enzymes that activate and deactivate a number of different metabolic pathways by catalyzing the phosphorylation of specific tyrosine residues on key effector proteins. Tyrosine kinases are typically classified into membrane-bound or non-membrane-bound groups and are found in nearly every mammalian cell type.

RTF siRNA libraries are provided as multiple single-use plate sets – just rehydrate, and add cells. The pre-plated format reduces hands-on time for faster screening results.

ON-TARGETplus modifications combine sense-strand inactivation with a novel seed-region modification. This patented siRNA approach is the best strategy to prevent off-target effects caused by both the sense and antisense strands while maintaining high-silencing potency.

Highlights

  • Six ready-to-use 96-well plate sets provided for two triplicate screens
  • Pre-plated, validated RNAi controls included
  • No aliquoting necessary, just resuspend and add cells
  • siRNA reagents provided in clear plates at 6.25 pmol per well (50 nM final screening concentration)
  • Black or white clear-bottom plates available to support assays involving fluorescent or luminescent detection
Well Pre-plated controls in ON-TARGETplus RTF Libraries
A1 ON-TARGETplus Non-targeting siRNA #1 D-001810-01
B1 ON-TARGETplus Non-targeting siRNA #2 D-001810-02
C1 ON-TARGETplus Non-targeting siRNA #3 D-001810-03
D1 ON-TARGETplus Non-targeting siRNA #4 D-001810-04
E1 ON-TARGETplus Non-targeting pool D-001810-10
1F ON-TARGETplus Human Cyclophilin B control pool D-001820-10

For a diagram of the RTF Library Plate layout, see Figure 1 on the Supporting Data tab.

Experimental considerations

DharmaFECT Cell Culture Reagent (DCCR) is recommended for use with RTF libraries to dilute transfection reagents prior to use. DCCR may be purchased separately or accompanying your RTF Optimization Plates

DharmaFECT transfection reagents are highly recommended for use with RTF libraries and should be purchased separately. Refer to the DharmaFECT Cell Type Guide to find the appropriate formulation for your cell type.

Gene Targets

For a complete list of target genes in this siRNA Library please email Technical Support or call 1-800-235-9880. International customers please call 303-604-9499 or your local Sales Representative.

  
HazardousNo
Shelf Life12 Months
Shipping ConditionAmbient
Storage Condition4 C

Our siRNA knockdown guarantee

siGENOME and ON-TARGETplus siRNA reagents (SMARTpool and three of four individual siRNAs) are guaranteed to silence target gene expression by at least 75% at the mRNA level when used under optimal delivery conditions (confirmed using validated positive control and measured at the mRNA level 24 to 48 hours after transfection using 100 nM siRNA).

Note: Most siGENOME and ON-TARGETplus siRNA products are highly functional at 5 to 25 nM working concentration.

RTF siRNA Library Plate Layout

RTF siRNA Library Plate Layout

RTF siRNA Library Plate Layout


Reverse Transfection Format is highly effective across cell lines

Reverse Transfection Format is highly effective across cell lines

Reverse Transfection Format is highly effective across cell lines

Eight cell lines were transfected with control siRNAs targeting Cyclophilin B using RTF plates. Good cellular viability and effective target silencing were accomplished under optimized conditions. Gene knockdown is normalized to GAPDH expression. Cell plating densities are indicated in parentheses. Gene expression was determined 48 hours posttransfection by branched DNA assay (Panomics Quantigene® Reagent System) (blue bars). Cell viability was determined by alamarBlue® metabolic assay (gold circles).


References

  1. General Screening References
  2. B. D. Parsons et al., A direct phenotypic comparison of siRNA pools and multiple individual duplexes in a functional assay. PLoS One4(12), e8471 (29 December 2009).
  3. M. Jiang et al., Tales from an academic RNAi screening facility; FAQs. Brief Funct Genomics. 10(4), 227-237 (Epub 28 April 2011, July 2011). [doi: 10.1093/bfgp/elr016]

Citations

  1. Publications using RTF Libraries
  2. T. Sorkina et al., RNA interference screen reveals an essential role of Nedd4-2 in dopamine transporter ubiquitination and endocytosis. J Neurosci26(31), 8195-8205 (2 August 2006).
  3. P. Monteiro et al., O. Dioxin-mediated up-regulation of aryl hydrocarbon receptor target genes is dependent on the calcium/calmodulin/CaMKIalpha pathway. Mol Pharmacol. 73(3), 769-777 (Epub 18 December 2007, March 2008).
  4. A. A. Kolokoltsov et al., Small interfering RNA profiling reveals key role of clathrin-mediated endocytosis and early endosome formation for infection by respiratory syncytial virus. J Virol. 81(14),7786-7800 (Epub 9 May 2007, July 2007).
  5. K. M. Hussain KM et al., The essential role of clathrin-mediated endocytosis in the infectious entry of human enterovirus 71. J Biol Chem286(1), 309-321 (Epub 18 October 2010, 7 January 2011).