( Proven siRNA Controls and Matche...)Control Your siRNA Research

Proven siRNA Controls and Matched Primary Antibodies

Gene Specific siRNA Controls
Whether you are starting out in siRNA research or you are an siRNA expert, you'll appreciate the convenience of the Silencer gene specific siRNA controls. Each Silencer gene specific siRNA control contains 50 g of ready-to-use chemically synthesized and gel purified GAPDH, cyclophilin, c-myc, or -actin siRNA, which is sufficient for 165 transfections (24 well plates). Also included are 25 g of a scrambled Negative Control siRNA and a detailed Instruction Manual. Each control is validated for use in human cell lines and each lot is functionally tested in an siRNA experiment. The GAPDH and cyclophilin siRNAs are also validated for use in mouse cell lines. These siRNA controls are ideal for developing and optimizing siRNA experiments.

Monitoring Transfection Efficiency
Because low transfection efficiency is the most frequent cause of unsuccessful gene silencing experiments, optimizing transfection conditions is critical. The Silencer Gene Specific siRNA Controls are ideal for this application, particularly when they are fluorescently labeled using the simple procedure provided with the Silencer siRNA Labeling Kits. Once transfected into cells, uptake of the labeled siRNA can be correlated to gene silencing by fluorescence microscopy using one of Ambion's new primary antibodies matched to our gene specific siRNA controls (Figure 1).

Figure 1. Characteristics of Ambion's Primary Antibodies for siRNA Research.

Primary Antibodies Matched to Silencer Gene Specific Controls
Ambion provides mouse monoclonal antibodies for the detection of GAPDH, c-myc, and -actin by immunofluorescence. Each of these primary antibodies has been used in immunofluorescence experiments at Ambion to detect the reduction in protein levels induced by siRNA. An example of such an experiment is shown in Figure 2. (Antibodies are supplied in solution in a 100 g unit size and are validated for use in immunofluorescence experiments. Note that the concentration and volume will vary by antibody).

Figure 2. Following the Silencing of -actin. An siRNA targeting -actin was labeled with Cy3 using the Silencer siRNA Labeling Kit. The labeled siRNA was transfected into HeLa cells and cells were analyzed 96 hours later. Green: -actin protein detected with anti--actin (Ambion) and a fluorescein labeled secondary antibody. Red: Cy3 labeled siRNA. Blue: DAPI stained nuclei. (Cy3 is a trademark of Amersham Biosciences.)

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Ordering Information
Cat# Product Name Size 1632 Silencer siRNA Labeling Kit - Cy3 65 g labeled siRNA 1634 Silencer siRNA Labeling Kit - FAM 65 g labeled siRNA 4300 anti-GAPDH, mouse monoclonal 6C5 100 g 4302 anti-beta-actin, mouse monoclonal AC-15 100 g
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