from Pete Shuster at Neuromics
Intraventricular Injections Used to Study Neuronal Survival During Post Natal Development
Neuromics’ i-Fect
TM Transfection Kit continue to be successfully used to deliver siRNA, shRNA and miRNA to cell cultures and the CNS
in vivo (via intrathecal, epidural and intraventricular injection). Genes studied include: ABCA, ASIC, β-arrestin, CAV
1.2, IGF1, CX3CR1, DOR, ELOVL4, IKBKAP, K+-ATPase, K
V1.1, K
V9.1 ,The β3 subunit of the Na+,K+-ATPase, NTS1, NAV
1.8, NTS1, NOV, Raf-1, RANK,SNSR1, hTertTRPV1 NOV, Survivin, TLR4, Troy and TRPV1.
Related Publications.
It is always an honor for one our products to be referenced in one of the
Nature publications. Here researchers use i-Fect to study the impact of microglial derived IGF1 silencing on Neuronal Survival:
Masaki
Ueno,Yuki Fujita, Tatsuhide Tanaka, Yuka Nakamura, Junichi Kikuta,
Masaru Ishii, Toshihide Yamashita. Layer V cortical neurons require
microglial support for survival during postnatal development. Nature Neuroscience 16, 543–551 (2013) doi:10.1038/nn.3358. …
vehicle
(PBS) was delivered intraventricularly through the cisterna magna with a
glass pipette while P3 mice were cold anesthetized. Igf1 siRNA (stealth
siRNA, Invitrogen) or Igfbp5 siRNA with i-Fect reagent (Neuromics) was
delivered intraventricularly through the cisterna magna at P3 twice with
a 12-h interval…
Images: (a) Igf1 expression (blue) and Iba1-positive microglia
(brown) in P5 brain. Scale bar represents 400 μm. (b) Magnified view of
dotted square in a. Scale bar represents 50 μm. (c,d) IGF1Ra expression
(red) in CTIP2-positive (c) and SATB2-positive (d) layer V neurons at
P5. Scale bar represents 50 μm. (e) IGF1 protein levels in medium from
cultured cortical neurons, microglia and neurons with microglia in
transwell systems detected by enzyme-linked immunosorbent assay (ELISA).
**P < 0.01 (neuron, microglia + neuron, n = 5; microglia, n = 6
experiments; one-way ANOVA followed by Tukey-Kramer test). (f) The
number of cleaved caspase-3–positive neurons in transwell systems
treated with LY294002 or H-1356 or transfected to microglia with Igf1
siRNA. *P < 0.05, **P less than 0.01 (n = 3 experiments, one-way
ANOVA followed by Tukey-Kramer test). (g) Neuronal phospho-AKT
expression in cultured cortical neurons and those with microglia in
transwell system. (h) TUNEL-positive cells in H-1356–treated cortex (36 h
after treatment). Scale bar represents 100 μm. (i) The number of
TUNEL-positive apoptotic cells in each layer in vehicle-
(phosphate-buffered saline, PBS) or H-1356–treated mice (36 h after
injection). *P < 0.05 (n = 4 brains, one-way ANOVA followed by
Tukey-Kramer test). (j) Cleaved caspase-3–labeled cells (red) expressing
CTIP2 (green, arrowheads). Scale bar represents 100 μm. (k,l)
TUNEL-positive cells in the cortex treated with control or Igf1 siRNA
(48 h after treatment). Scale bar represents 100 μm. (m) The number of
TUNEL-positive apoptotic cells in each layer in control siRNA– and Igf1
siRNA–treated mice. **P less than 0.01 (n = 5 brains, one-way ANOVA
followed by Tukey-Kramer test). Error bars represent s.e.m.
Related Products
i-Fect
| Catalog Number |
Size |
|
|
| NI35150 |
1 x 0.75 ml |
|
|
| NI35750 |
5 x 0.75 ml |
|
|
|
Type: Cationic Lipid
Description/Data:
Figure:
siRNA-mediated suppression of target gene expression in Schwann cells.
A,
Detergent extracts of siNeg- or siGly1-transfected Schwann cells were
digested with heparitinase and subjected to immunoblot analysis with
anti-glypican-1 antibodies (top); aliquots of undigested extracts were
immunoblotted with anti-actin antibodies (bottom) to verify equal sample
loading.
B, Cell surface expression of glypican-1 was assessed
by immunofluorescent staining of transfected cells 48 h after
transfection using anti-glypican-1 antibodies (green) and DAPI
(4′,6′-diamidino-2-phenylindole) to stain nuclei (red).
C,
Schwann cells were transfected with siNeg or si alpha4(V), and
conditioned media and cell lysates were harvested 48 h later (left) or
at the indicated times after transfection (right); aliquots of medium
(top) or cell lysates (bottom) were subjected to immunoblot analysis and
stained with anti-alpha4(V) collagen (top) or anti-actin (bottom)
antibodies.
i-Fect
TM is a novel cationic lipid formulation
specifically designed for efficient delivery of siRNAs (small
interfering RNAs) into a wide variety of cell types. Customers have
been using i-Fect for both
in vitro and
in vivo applications.
Kit Contents
-
Hydrated i-Fect ™ Lipid
-
siRNA Diluent
Protocol
i-Fect ™ Hi Put 96-New
i-Fect based system for high throughput delivery of siRNAs (small interfering RNAs) into a wide variety of cell types.
Image: 96 well plate showing initial dilution series for optimizing siRNA delivery.
i-Fect ™ has proven to be an effective tool for delivering siRNA and DsiRNA
in vitro and
in vivo. These capablities are confirmed in a growing list of
Publications referencing i-Fect.
i-Fect Hi-Put 96 provides a convenient high-throughput format for
optimizing transfection conditions and performing siRNA transfection in
96-well plates. All wells (except those in Column 12) contain a
predetermined amount of the i-Fect siRNA Transfection Reagent, a
cationic lipid based reagent that has been extensively screened in many
mammalian cell lines in order to achieve:
- Optimal dilutions for high percentage delivery of siRNA
- Functional gene silencing post siRNA delivery.
- Compatibility with diverse growth conditions (with and without serum).
- Low cytotoxicity.
| Name |
CatalogUE |
Type |
Species |
Applications |
Size |
|
| 96 Well Dilution Series |
PL30001 |
Cationic Lipid |
|
|
96 Well Tritration Plate
4X96 Well Tritration Plate |
|
| 96 Well (High) |
PL30002 |
Cationic Lipid |
|
|
96 Well Plate-High
4X96 Well Plate-High |
|
| 96 Well (Low) |
PL30003 |
Cationic Lipid |
|
|
96 Well Plate (Low)
4X96 Well Plate (Low) |
|
| siRNA-intact |
SI35100 |
RNAse Inhibitor |
|
|
100 units in 100 ul |
|
Where can I find more information about Neuromics?
Visit the manufacturer page at
www.stratech.co.uk/neuromics, email
info@stratech.co.uk or call +44 (0) 1638 782600.
Stratech Scientific is a distributor of high quality, competitively
priced, reliable products for research laboratories throughout the UK
and Europe.
Please contact us to find out which product ranges we can supply in your country.
