Smartpool

Manufactured by Thermo Fisher Scientific

Sourced in United States

SMARTpool is a pre-designed and pre-packaged set of gene-specific siRNA reagents from Thermo Fisher Scientific. It is designed to provide effective gene knockdown by targeting multiple sites within a gene sequence.

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Lab products found in correlation

46 protocols using smartpool

Modulating FILIP1L and SLUG in Ovarian Cancer

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ON-TARGETplus Non-Targeting siRNA Pool (D-001810-10), FILIP1L siRNA (#7 (J-019458-07) and SMARTpool (set of 4 sequences; L-019458-00)) and SLUG siRNA (#8 (J-017386-08) and SMARTpool (L-017386-00)) were purchased from Thermo Scientific. HEY and OVCAR8 cells were transfected with equimolar amounts of either non-Targeting or FILIP1L and/or SLUG siRNA using Dharmafect solution following the manufacturer's protocols (Thermo Scientific). After a 48 h transfection, the cell lysates were subjected to qRT-PCR and immunoblot analysis.

Kwon M., Kim J.H., Rybak Y., Luna A., Choi C.H., Chung J.Y., Hewitt S.M., Adem A., Tubridy E., Lin J, & Libutti S.K. (2016). Reduced expression of FILIP1L, a novel WNT pathway inhibitor, is associated with poor survival, progression and chemoresistance in ovarian cancer. Oncotarget, 7(47), 77052-77070.

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Transient Transfection of SUV39H1 and BRCA1 siRNA

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For transient transfection, human SUV39H1 siRNA was purchased from Thermo Scientific (SMARTpool, ON-TARGETplus), and the SUV39H1 target sequences were CUAAGAAGCGGGUCCGUAU (J-009604-07), GGUGAAAUGGCGUGGAUAU (J-009604-08), UCGAGUACCUGUGCGAUUA (J-009604-09), and CAAAUCGUGUGGUACAGAA (J-009604-10). Human BRCA1 siRNA was purchased from Thermo Scientific (SMARTpool, ON-TARGETplus), and the BRCA1 target sequence was CAACAUGCCCACAGAUCAA (J-003461-09), CCAAAGCGAGCAAGAGAAU (J-003461-10), UGAUAAAGCUCCAGCAGGA (J-003461-11), and GAAGGAGCUUUCAUCAUUC (J-003461-12). All siRNAs were transfected with oligofectamine for 48-72 hours for further analysis.

Mo W., Liu Q., Lin C.C., Dai H., Peng Y., Liang Y., Peng G., Meric-Bernstam F., Mills G.B., Li K, & Lin S.Y. (2015). mTOR inhibitors suppress homologous recombination repair and synergize with PARP inhibitors via regulating SUV39H1 in BRCA-proficient triple-negative breast cancer. Clinical cancer research : an official journal of the American Association for Cancer Research, 22(7), 1699-1712.

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Silencing STAMP2 and FSP27 in Rat Cells

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Rat STAMP2 siRNA (SMARTpool; L-105419-02-0050) and FSP27 siRNA (SMARTpool; L-105647-02-0050) were purchased from Thermo Scientific (Hudson, NH, USA). As a negative control, the same nucleotides were scrambled to form nongenomic combinations. Transfection of the siRNA was performed with the use of siPORT Amine and Opti-MEM medium. Cells grown to a confluency of 40%–50% in six-well plates were transfected with 100 nmol·L^-1 of siRNA per well. The transfection mixture was added to each well, and the cells were incubated for 4 h. Then, 2 mL of growth medium was added, and the cells were incubated for another 20 h. After the siRNA transfection medium was removed, cells were treated with FFAs for 24 h.

Lee S.W., Rho J.H., Lee S.Y., Chung W.T., Oh Y.J., Kim J.H., Yoo S.H., Kwon W.Y., Bae J.Y., Seo S.Y., Sun H., Kim H.Y, & Yoo Y.H. (2018). Dietary fat-associated osteoarthritic chondrocytes gain resistance to lipotoxicity through PKCK2/STAMP2/FSP27. Bone Research, 6, 20.

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Silencing BCL2L11 and CDH9 in Breast Cancer Cells

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Smartpool: on-targetplus BCL2L11, CDH9 and control siRNA were purchased from Dharmacon.
Human BCL2L11 siRNA - Smartpool, L-004383-00-0005
Human CDH9 siRNA - Smartpool, L-013169-00-0005
Non-targeting Pool, D-001810-10-05
SiRNAs were transfected into HCC1937, MDA-MB-231, and HCC1806 cells using Lipofectamine RNAiMAX Reagent (Life technologies).

Yan G., Chen V., Lu X, & Lu S. (2017). A signal-based method for finding driver modules of breast cancer metastasis to the lung. Scientific Reports, 7, 10023.

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Silencing Usp9X, Bag-3, and Mcl-1 in Cells

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SignalSilence^® Usp9X siRNA I #6308 was purchased from CST. Non-targeting siRNA-pool (ON-TARGETplus Non-targeting Pool, # D-001810-10-05) and siRNA against Bag-3 (SMARTpool: ON-TARGETplus Bag3 siRNA, L-011957-00-0005) and Mcl-1 (SMARTpool: ON-TARGETplus Mcl-1 siRNA, L-004501-00-0005) were purchased from Thermo Fisher Scientific (Pittsburgh, PA) and transfected as previously described [38 (link)].

Karpel-Massler G., Shu C., Chau L., Banu M., Halatsch M.E., Westhoff M.A., Ramirez Y., Ross A.H., Bruce J.N., Canoll P, & Siegelin M.D. (2015). Combined inhibition of Bcl-2/Bcl-xL and Usp9X/Bag3 overcomes apoptotic resistance in glioblastoma in vitro and in vivo. Oncotarget, 6(16), 14507-14521.

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Transient Silencing of EGFR in MMECs

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MMECs (4 × 10⁵ cells/100 mm Petri dish) were transiently transfected with 50 nM EGFR siRNA (small interfering RNA) or control non-targeting siRNA (SMARTpool; Thermo Fisher) using the transfection reagent Lipofectamine (Life Technologies) for 48 h. siRNA-transfected MMEC were evaluated immediately in western blot and angiogenesis assays.

Rao L., Giannico D., Leone P., Solimando A.G., Maiorano E., Caporusso C., Duda L., Tamma R., Mallamaci R., Susca N., Buonavoglia A., Da Vià M.C., Ribatti D., De Re V., Vacca A, & Racanelli V. (2020). HB-EGF–EGFR Signaling in Bone Marrow Endothelial Cells Mediates Angiogenesis Associated with Multiple Myeloma. Cancers, 12(1), 173.

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Macrophage Differentiation and Transfection

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Freshly isolated PBMCs were differentiated to macrophages using 6-day differentiation in 10% human serum supplemented with 5 ng/mL GM-CSF (R&D systems). After differentiation (1 × 10⁵) macrophages were seeded in 96 well plates and left for 2 h at 37 °C to subsequently transfect them with 25 nM NOD2 targeting siRNA (on target) or scrambled (non-target) control siRNA (smart pool, Thermo Scientific) for 24 h at 37 °C (Dharmafect, Thermo Scientific). Subsequently, the culture medium was refreshed and cells were used for killing and phagocytosis assays and PCR analysis.

Gresnigt M.S., Cunha C., Jaeger M., Gonçalves S.M., Malireddi R.K., Ammerdorffer A., Lubbers R., Oosting M., Rasid O., Jouvion G., Fitting C., Jong D.J., Lacerda J.F., Campos A J.r., Melchers W.J., Lagrou K., Maertens J., Kanneganti T.D., Carvalho A., Ibrahim-Granet O, & van de Veerdonk F.L. (2018). Genetic deficiency of NOD2 confers resistance to invasive aspergillosis. Nature Communications, 9, 2636.

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Modulating miR-361-5p and UBR5 in Glioblastoma

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miR-361-5p mimic and inhibitor sequence and its corresponding negative control (NC) sequence dsRNA oligonucleotides were available from Ribobio (Guangzhou, China). U87 and U251 cells growing to 50% confluence were transfected with corresponding dsRNA oligonucleotides (2 μg) through 10 μL X-tremeGENE siRNA Transfection Reagent (Hoffmann-La Roche Ltd., Basel, Switzerland). The final concentration in transfection was 50 nM [7 (link)]. The siRNA Smartpool and RISCfree control siRNA against human and mouse UBR5 (Thermo Fisher Scientific, Massachusetts, USA) were transfected into U87 and U251 cells through Dharmafect 1 (Invitrogen) [13 (link)].

Jia J., Ouyang Z., Wang M., Ma W., Liu M., Zhang M, & Yu M. (2021). MicroRNA-361-5p slows down gliomas development through regulating UBR5 to elevate ATMIN protein expression. Cell Death & Disease, 12(8), 746.

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Modulation of Candida-induced IL-6 in PBMCs

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Peripheral blood mononuclear cells (PBMC) were isolated from healthy volunteers by Ficoll-Paque gradient. After isolation 0.5 × 10⁶ cells were plated into 96 round bottom well-plates and left for 2 h at 37°C to subsequently transfect them with 25 nM NT5C3, CD38 siRNA (on target) or scrambled (non target) control siRNA (smartpool, Thermo Scientific) for 48 h at 37°C (Dharmafect, Thermo Scientific). After a centrifugation step (8 min 1800 rpm), supernatant was discarded and cell pellet was resuspended in 200 μl of RPMI (Dutch modified) containing Candida albicans blastoconidia or Candida hyphae (both UC820, 1 × 10⁶/ml, heat-killed at 100°C for 1 h) for 24 h. Supernatants were collected and IL-6 was subsequently measured by enzyme-linked immunosorbent assay (ELISA) (Sanquin Research, Amsterdam).

Goel G., Conway K.L., Jaeger M., Netea M.G, & Xavier R.J. (2014). Multivariate inference of pathway activity in host immunity and response to therapeutics. Nucleic Acids Research, 42(16), 10288-10306.

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Silencing CK2 and PKC in HepG2 Cells

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Silencing of CK2 holoenzyme in HepG2 cells was achieved using SMART pool (Thermo Scientific, Rockford, IL, USA) siRNA (100nM) targeting all three subunits of human CK2 (α, α’ and β) as described previously (Abu Shehab et al 2014 (link)). PKC was silenced using pan-PKC (100 nM siRNA) (targeting PKC isoforms α, β, βII, γ, δ, ε, η, θ, ζ, and ν) (Santa Cruz Biotechnology, Dallas, TX, USA). Transfection was performed using Dharmacon transfection reagent 4 (Dharmacon,Thermo Fisher Scientific, Waltham, MA). Transfection media was removed after 24 hours. HepG2 cells were then incubated with leucine plus or leucine minus media for an additional 72 hours (Figure S1). Western immunoblot analysis was performed using CK2α, CK2α’, CK2β and PKCδ and PKCε antibodies to validate silencing efficiency.

Malkani N., Biggar K., Shehab M.A., Li S., Jansson T, & Gupta M.B. (2015). Increased IGFBP-1 phosphorylation in response to leucine deprivation is mediated by CK2 and PKC. Molecular and cellular endocrinology, 425, 48-60.

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