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).
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).
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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Ordering Information
Cat#
Product Name
Size
1632
Silencer® siRNA Labeling Kit - Cy™3
65 µg labeled siRNA
1634
Silencer® siRNA Labeling Kit - FAM
65 µg labeled siRNA
4300
anti-GAPDH, mouse monoclonal
6C5
100 µg
4302
anti-beta-actin, mouse monoclonal AC-15
100 µg