Cat.-no	Description	Amount	Price €	Shop
70500	siRNA-Trans Maximo	500 µl	199.00	add
71000	siRNA-Trans Maximo	1 ml	349.00	add
73000	siRNA-Trans Maximo	3x1 ml	949.00	add

siRNA-Transfection reagent: Manual

Exceptional protein expression

siRNA Trans Reagent is the ideal siRNA transfection reagent for gene silencing. It has been successfully tested on various cell lines, reaching up to 90% gene silencing. Due to its properties, siRNA Trans Reagent is a very efficient reagent leading to exceptional knockdown effects with low doses of siRNA. It protects siRNA from extracellular degradation and has an outstanding ability to destabilize cell membranes. Consequently it allows reproducible delivery of important siRNA amounts into the cytosol and high cell viability is maintained in each experiment.

• Effective with very low doses of siRNA
• Minimize off-target effects
• Perfect for endogenous gene targeting and co-transfection
• Applicable to a broad range of cells
• Serum compatible & Non toxic

High reliable and reproducible gene silencing

siRNA Trans Reagent is a very efficient siRNA delivery reagent which leads to reproducible protein knockdown with no off-target effects.
 
Great protein knockdown with low doses of siRNA

GFP inhibition in HeLa cells









Dose response GFP inhibition in HeLa cells




Gene silencing in various cell lines



Endogenous protein shutdown

TAK1 kinase inhibition in human pancreatic cells











GAPDH inhibition in HEK cells










siRNA transfection reagents comparison

GFP knockdown compared with competitors

Troubleshooting

 

Problems	Comments and Suggestions
Low transfection efficiency	1- siRNA-Trans reagent / siRNA ratio. Although our reagent has been extensively optimized and fixed volumes are provided, optimization may be required. Optimize the Lullaby/siRNA ratio as described in the optimization protocol. Briefly, use a fixed amount of siRNA (10 or 20nM) and vary the amount of siRNA-Trans reagent from 2 times less up to three times more than the suggested amount detailed in the Table 3 2- siRNA amount. Use different concentration of siRNA with the recommended or optimized Lullaby/siRNA ratio. 3- Cell density. A non-optimal cell density at the time of siRNA delivery can lead to insufficient uptake. The optimal confluency should range from 50 to 70% but most favorable cell density may vary according to the cell type. 4- siRNA quality. Use high quality siRNA (PAGE purified and desalted). Employ RNAse-free materials and check for siRNA integrity on acrylamide gel. Ensure siRNA is not denatured. 100mM NaCl, 50mM Tris pH7.5 RNAse-free buffer can be used for siRNA instead of water. 5- Mycoplasma contamination. Mycoplasma contamination alters transfection efficiency. 6- Cell condition. Cells that have been in culture for a long time may become resistant to transfection. Use freshly thawed cells that have been passaged at least once.
Cellular toxicity	1- Concentration of siRNA-TRANS Maximo reagent  / siRNA. Decrease the amount of siRNA / reagent complexes added to the cells. 2- Incubation time. Reduce the incubation time of complexes with the cells. Transfection medium can be replaced by fresh medium after 4h. 3- siRNA quality. Use high quality siRNA as impurities can lead to cell death. 4- Key gene silencing. If the targeted gene is essential for cell survival or if a key gene is non-specifically silenced by the siRNA this can lead to cell death 5- Unhealthy cells. Check cells for contamination, use new batch of cells, ensure culture medium condition (pH, type of medium used, contamination etc)
No or weak gene silencing effect	1- siRNA design. The design of an efficient siRNA is a crucial step. Ensure to use a validated siRNA sequence. If a validated siRNA cannot be used, assay your sequence in an easy to transfect cell line (if possible) in order to validate it (HeLa cells for example). 2- siRNA concentration. Use higher amount of siRNA. 3- Incubation time. Perform a time-course experiment to set up the optimal incubation time since gene silencing is dependent on the gene expression and the protein turnover rate. 4- Incorrect medium used for preparing siRNA-Trans/siRNA complexes. It is critical that serum-free medium or buffer (HBS, PBS) are used during the preparation of the Lullaby/siRNA complexes. 5- Old siRNA-Trans/siRNA complexes. The Lullaby/siRNA complexes must be freshly prepared every time. Complexes prepared and store for longer than 1 hour can be aggregated. 6- Improper storage. siRNA-TRANS Maximo reagent  is very stable at 4°C but high temperature and/or excessive freeze/thaw cycles may cause lost of reagent activity.

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