Stable HeLa cell lines expressing GFP‐α‐tubulin and RFP‐Lifeact were established starting from HeLa stably expressing GFP‐α‐tubulin by lentiviral transduction with rLV‐Lifeact‐RFP (Ibidi) using 3 MOI (multiplicity of infection). After 24 h, cells were incubated with fresh medium for 48 h. After 72 h post‐transduction, stable cells were selected using 1 μg/ml puromycin. Medium with puromycin was replaced every 2–3 days until resistant colonies were identified. ON‐TARGETplus Human SMARTpool WASH1‐targeting (siWASH1) siRNAs (Dharmacon, GE Healthcare) were transfected into HeLa cells at a final concentration of 20 nM using Lipofectamine RNAiMax (Life Technologies) according to supplier's protocol. Negative control siRNA was performed using AllStars Negative Control siRNA (Qiagen).
On targetplus human smartpool wash1 targeting siwash1 sirnas
Manufactured by Horizon Discovery
ON-TARGETplus Human SMARTpool WASH1-targeting (siWASH1) siRNAs are a set of four individual siRNA duplexes designed to target the WASH1 gene in human cells. These siRNAs are pre-designed and pre-assembled into a single pool to facilitate efficient gene silencing.
Automatically generated - may contain errors
Lab products found in correlation
Lipofectamine rnaimax, by Thermo Fisher Scientific (3 mentions)
Lipofectamine 2000, by GE Healthcare (2 mentions)
Geneticin, by Thermo Fisher Scientific (2 mentions)
Sipcm1, by Qiagen (2 mentions)
Lipofectamine 2000, by Thermo Fisher Scientific (2 mentions)
X tremegene dna transfection reagent, by Roche (2 mentions)
3 protocols using on targetplus human smartpool wash1 targeting siwash1 sirnas
1
Establishment and Manipulation of Stable HeLa Cell Lines
2
Establishment of EGFP-Centrin1 Expressing Jurkat Cells
3
Establishment of EGFP-Centrin1 Expressing Jurkat Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
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Stable Jurkat cell lines expressing EGFP-centrin1 were established by transfecting cells with pEGFP-centrin1 using Lipofectamine 2000 (Life Technologies) at a ratio of 2.5:1 transfection reagent (μl) to DNA (μg). Medium was supplemented with 0.75 mg/ml geneticin (Gibco) 24 hours after transfection to select for cells stably incorporating the EGFP-centrin1 sequence. After 2 weeks of selection, fluorescent-activated cell sorting was carried out to eliminate non-expressing cells from the culture. Transient transfections in Jurkat and HEK293T cells were performed using X-tremeGENE DNA transfection reagent (Roche) according to supplier’s protocol using a 3:1 ratio of transfection reagent (μl) to DNA (μg). Small interfering RNAs (siRNAs) targeting PCM1 (siPCM1, Qiagen) and ON-TARGETplus Human SMARTpool WASH1-targeting (siWASH1) siRNAs (Dharmacon, GE Healthcare) were transfected into Jurkat cells at the final concentration of 20 nM using Lipofectamine 2000 according to supplier’s protocol. PCM1-targeting siRNAs were transfected into RPE1 cells at the final concentration of 10 nM using Lipofectamine RNAiMax (Life Technologies) according to supplier’s protocol. Negative control siRNA was performed using AllStars Negative Control siRNA (Qiagen).
PCM1-target sequences were:
PCM1-target sequences were:
HS-PCM-1_1 5′-CAGUAUCACAUCUGAACUAAA-3′
HS PCM-1_2 5′-CAGGCUUUAACUAAUUAUGGA-3′
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