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 µg
of ready-to-use chemically synthesized and gel purified GAPDH, cyclophilin,
c-myc, or ß-actin siRNA, which is sufficient for 165 transfections
(24 well plates). Also included are 25 µg 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).
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| 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 ß-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 µg unit size
and are validated for use in immunofluorescence experiments. Note that the
concentration and
volume will vary by antibody).
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| Figure 2. Following
the Silencing of ß-actin. An
siRNA targeting ß-actin was labeled with Cy™3 using
the Silencer™ siRNA Labeling Kit. The labeled siRNA
was transfected into HeLa cells and cells were analyzed
96 hours later. Green: ß-actin protein detected with
anti-ß-actin (Ambion) and a fluorescein labeled secondary
antibody. Red: Cy3 labeled siRNA. Blue: DAPI stained nuclei.
(Cy™3 is a trademark of Amersham Biosciences.) |
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