SANTA CRUZ BIOTECHNOLOGY, INC.
Slit2 siRNA (h): sc-42258
The Power to Question
BACKGROUND STORAGE AND RESUSPENSION
Secreted leucine-rich repeat-containing proteins 1-3 (Slit1-3) are secreted Store lyophilized siRNA duplex at -20° C with desiccant. Stable for at least
glycoproteins that influence axonal guidance and mediate normal neural one year from the date of shipment. Once resuspended, store at -20° C,
development by acting as high-affinity signaling ligands for the repulsive avoid contact with RNases and repeated freeze thaw cycles.
guidance receptor, Roundabout (Robo). Within the developing central nervous Resuspend lyophilized siRNA duplex in 330 µl of the RNAse-free water
system (CNS) of different vertebrate systems, Slit proteins are expressed in provided. Resuspension of the siRNA duplex in 330 µl of RNAse-free water
equivalent regions, suggesting a conserved function among vertebrate makes a 10 µM solution in a 10 µM Tris-HCl, pH 8.0, 20 mM NaCl, 1 mM
homologs. Slit is expressed in the midline of the central nervous system in EDTA buffered solution.
both vertebrates and invertebrates, where it functions as a regulatory factor
of mesodermal cell movement during gastrulation. Slit2 is a short range APPLICATIONS
inhibitory guidance cue for retinal ganglion cell (RGC) axons that may mediate
spatial progression of RGCs. Slit2 siRNA (h) is recommended for the inhibition of Slit2 expression in
REFERENCES Slit2 (E-20): sc-16619 is recommended as a control antibody for Western
1. Rothberg, J.M., et al. 1990. slit: an extracellular protein necessary for Blotting (starting dilution 1:100, dilution range 1:100-1:1,000) or immuno-
development of midline glia and commissural axon pathways contains fluorescence (starting dilution 1:50, dilution range 1:50-1:500) protein
both EGF and LRR domains. Genes Dev. 4: 2169-2187. detection using the recommended secondary reagents listed below.
2. Holmes, G.P., et al. 1998. Distinct but overlapping expression patterns of RECOMMENDED SECONDARY REAGENTS
two vertebrate slit homologs implies functional roles in CNS development
and organogenesis. Mech. Dev. 79: 57-72. To ensure optimal results, the following support (secondary) reagents are
3. Brose, K., et al. 1999. Slit proteins bind Robo receptors and have an evolu- recommended: 1) Western Blotting: use donkey anti-goat IgG-HRP: sc-2020
tionarily conserved role in repulsive axon guidance. Cell 96: 795-806. (dilution range: 1:2000-1:100,000) or Cruz Marker™ compatible donkey
anti-goat IgG-HRP: sc-2033 (dilution range: 1:2000-1:5000), Cruz Marker™
4. Hu, H. 1999. Chemorepulsion of neuronal migration by Slit2 in the devel- Molecular Weight Standards: sc-2035, TBS Blotto A Blocking Reagent:
oping mammalian forebrain. Neuron 23: 703-711. sc-2333 and Western Blotting Luminol Reagent: sc-2048. 2) Immunofluo-
5. Yuan, W., et al. 1999. The mouse SLIT family: secreted ligands for ROBO rescence: use donkey anti-goat IgG-FITC: sc-2024 (dilution range: 1:100-
expressed in patterns that suggest a role in morphogenesis and axon 1:400) or donkey anti-goat IgG-TR: sc-2783 (dilution range: 1:100-1:400)
guidance. Dev. Biol. 212: 290-306. with UltraCruz™ Mounting Medium: sc-24941.
6. Erskine, L., et al. 2000. Retinal ganglion cell axon guidance in the mouse SUPPORT REAGENTS
optic chiasm: expression and function of robos and slits. J. Neurosci. 20: For optimal siRNA transfection efficiency, Santa Cruz Biotechnology’s
4975-4982. siRNA Transfection Reagent: sc-29528 (0.3 ml), siRNA Transfection
7. Niclou, S.P., et al. 2000. Slit2 is a repellent for retinal ganglion cell axons. Medium: sc-36868 (20 ml) and siRNA Dilution Buffer: sc-29527 (1.5 ml)
J. Neurosci. 20: 4962-4974. are recommended. Control siRNAs or Fluorescein Conjugated Control
siRNAs are available as 10µM in 60 µl. Each contain a scrambled sequence
CHROMOSOMAL LOCATION that will not lead to the specific degradation of any known cellular mRNA.
Genetic locus: SLIT2 (human) mapping to 4p15.2; Slit2 (mouse) mapping to Fluorescein Conjugated Control siRNAs include: sc-36869, sc-44239,
5 B3. sc-44240 and sc-44241. Control siRNAs include: sc-37007, sc-44230,
sc-44231, sc-44232, sc-44233, sc-44234, sc-44235, sc-44236, sc-44237 and
PRODUCT sc-44238. Semi-quantitative RT-PCR may be performed using RT-PCR
Primer: Slit2 (h)-PR: sc-42258-PR (20 µl, 464 bp). Annealing temperature
Slit2 siRNA (h) is a target-specific 20-25 nt siRNA designed to knock down for the primers should be 55-60° C and the extension temperature should
gene expression. Each vial contains 3 nmoles of lyophilized siRNA, sufficient be 68-72° C.
for a 10 µM solution once resuspended using protocol below. Suitable for
For research use only, not for use in diagnostic procedures.
See our web site at www.scbt.com or our catalog for detailed protocols and
Santa Cruz Biotechnology, Inc. 1.800.457.3801 831.457.3800 fax 831.457.3801 Europe +00800 4573 8000 49 6221 4503 0 www.scbt.com
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