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Silencer select validated sirna for abcc4 silencing

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Silencer Select Validated siRNA for ABCC4 silencing is a laboratory product designed to reduce the expression of the ABCC4 gene in cell-based experiments. It provides a pre-designed and validated small interfering RNA (siRNA) sequence to target and silence the ABCC4 gene.

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Lab products found in correlation

Nucleofector 2, by Lonza (2 mentions) Nucleofector kit t, by Lonza (2 mentions) Trizol, by Thermo Fisher Scientific (2 mentions)

Close spelling variants of this product

Silencer® Select Validated siRNA Validated select siRNA Silencer Select siRNAs Silencer Select Silencer siRNA SiRNAs

2 protocols using silencer select validated sirna for abcc4 silencing

1

ABCC4 Gene Silencing in AML Cells

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The Silencer Select Validated siRNA for ABCC4 silencing (Ambion, Austin, TX) and the Silencer Select Negative Control #1 siRNA were used in all experiments. Cells were transfected with ABCC4 siRNA using Nucleofector II and nucleofector Kit T (Amaxa), according to the manufacturer's protocol. OCI‐AML3 cells (3 × 106) were treated with 100 μL reagent of solution T for each sample, using 1–3 μg of siRNA for each sample to optimize the best silencing effect. To evaluate ABCC4 suppression (see below), total RNA was extracted using TRIzol (Invitrogen) and analyzed by real‐time polymerase chain reaction (PCR) at variable times posttransfection. cDNA was synthesized by using a primer set designed to amplify exon 3, which encompasses the siRNA cleavage point. In each sample, the copy number of ABCC4 relative to glyceraldehydes‐3‐phosphate dehydrogenase (GAPDH) expression was calculated as follows: 2^dCt [dCt = Ct(GAPDH)‐Ct(ABCC4)]. Protein expression of ABCC4 was determined by western blot as described above.
Drenberg C., Hu S., Li L., Buelow D., Orwick S., Gibson A., Schuetz J., Sparreboom A, & Baker S. (2016). ABCC4 Is a Determinant of Cytarabine‐Induced Cytotoxicity and Myelosuppression. Clinical and Translational Science, 9(1), 51-59.
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2

Validating ABCC4 siRNA Silencing

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The Silencer Select Validated siRNA for ABCC4 silencing (Ambion) and the Silencer Select Negative Control #1 siRNA were used in all experiments. Cells were transfected with ABCC4 siRNA using Nucleofector II and nucleofector Kit T (Amaxa), according to the manufacturer’s protocol. OCI-AML3 cells (3×106) were treated with 100 μl reagent of solution T for each sample, using 1–3 μg of siRNA for each sample to optimize the best silencing effect. To evaluate ABCC4 suppression (see below), total RNA was extracted using TRIzol (Invitrogen) and analysed by real-time PCR at variable times post-transfection. cDNA was synthesized by using a primer set designed to amplify exon 3 which encompasses the siRNA cleavage point. In each sample, the copy number of ABCC4 relative to glyceraldehydes-3-phosphate dehydrogenase (GAPDH) expression was calculated as follows: 2^dCt [dCt=Ct(GAPDH)-Ct(ABCC4)]. Protein expression of ABCC4 was determined by western blot as described above.
Drenberg C., Hu S., Li L., Buelow D., Orwick S., Gibson A., Schuetz J., Sparreboom A, & Baker S. (2016). ABCC4 Is a Determinant of Cytarabine‐Induced Cytotoxicity and Myelosuppression. Clinical and Translational Science, 9(1), 51-59.
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