siRNA/miRNA Transfection Reagent

MedChemExpress Validation
Experimental conditions:
Cell density: 40% density during cell transfection
Dosage: 1 μL siRNA (EGFP/NC, 20 μM) + 2 μL transfection reagent (HY-K2017)
Detection scheme: 36 h after siRNA transfection, use fluorescence microscopy to detect the interference effect of EGFP green fluorescent protein
siRNA/miRNA Transfection Reagent (HY-K2017) demonstrated ​​high siRNA transfection efficiency and significant EGFP silencing in both A549 and HT29 cells.​

MedChemExpress Validation
Experimental Conditions:
Cell Density: 40% of the cell density at the time of transfection
Dosage: 2 μL siRNA (MALT1/NC, 20 μM) + 4 μL transfection reagent
Assay: 36 hours after siRNA transfection, RNA was extracted using Trizol and reverse transcribed, followed by qPCR verification of gene silencing efficacy.
The results demonstrate that the ​​siRNA/miRNA Transfection Reagent (HY-K2017)​​ achieves ​​significant silencing efficacy​​ against the ​​MALT1 gene​​ in ​​Caco-2 cells​​.

MedChemExpress Validation
GAPDH siRNA was transfected into 15 different cell types. At 36 h post-transfection, the remaining GAPDH mRNA levels were quantified by qPCR and normalized to the negative control (siNC), which was set to 1. The results showed that MCE siRNA/miRNA Transfection Reagent (HY-K2017) achieved significant mRNA knockdown across all 15 cell types tested. Robust silencing efficiency was maintained even in hard-to-transfect cells, including primary cells, neuronal cells, blood cells, endothelial cells, and various tumor cell lines.

MedChemExpress Validation
GAPDH siRNA (20 pmol) was transfected into HeLa, RKO, and H1299 cells using the siRNA/miRNA Transfection Reagent (HY-K2017). At 48 h post-transfection, cells were lysed and GAPDH protein levels were analyzed by Western blot, with β-actin used as the internal control. The results showed that efficient mRNA silencing was successfully translated into marked protein-level reduction, with GAPDH knockdown efficiencies of 85% in HeLa cells, 91% in H1299 cells, and up to 94% in RKO cells.

MedChemExpress Validation
GAPDH siRNA (20 pmol) was transfected into HT29 cells using the siRNA/miRNA Transfection Reagent (HY-K2017) or the competitor RNAiMAX. At 48 h post-transfection, cells were lysed and GAPDH protein levels were analyzed by Western blot, with β-actin as the internal control. Western blot analysis showed that MCE siRNA/miRNA Transfection Reagent mediated a significantly greater reduction in GAPDH protein levels compared with the competitor reagent.

MedChemExpress Validation
The siRNA/miRNA Transfection Reagent (HY-K2017) was used to deliver siRNA targeting the luciferase gene in HeLa, MDA-MB-231, HCT116, and NIH/3T3 cells. Relative luciferase activity was measured at 48 h post-transfection, with siNC as the control. MCE siRNA/miRNA Transfection Reagent (HY-K2017) achieved significant luciferase knockdown across all tested cell lines, including HeLa, MDA-MB-231, HCT116, and NIH/3T3.

MedChemExpress Validation
The siRNA/miRNA Transfection Reagent (HY-K2017) was used to deliver 10, 20, 40, and 60 pmol of siRNA targeting the luciferase gene in HeLa cells. Relative luciferase activity was measured at 48 h post-transfection, with siNC as the control. Efficient, dose-dependent gene silencing was observed across the siRNA input range of 10-60 pmol.

MedChemExpress Validation
The siRNA/miRNA Transfection Reagent (HY-K2017) was used to deliver siRNA targeting EGFP in MCF-7 cells stably expressing EGFP. At 48 h post-transfection, EGFP fluorescence intensity was quantified by flow cytometry. The mean fluorescence intensity (MFI) of EGFP-positive cells was analyzed, with siNC as the control, to evaluate the silencing efficiency of siEGFP. Quantitative flow cytometry analysis demonstrated a significant reduction in mean fluorescence intensity in EGFP-expressing MCF-7 cells following siRNA/miRNA transfection. The high-throughput and high-sensitivity detection further confirmed the reliability and reproducibility of MCE siRNA/miRNA Transfection Reagent (HY-K2017).

MedChemExpress Validation
Using the siRNA/miRNA Transfection Reagent (HY-K2017), Control ASO (negative control) and Luc ASO (luciferase-targeting antisense oligonucleotide) were transfected into MCF-7, HCT116, and HeLa cells. At 48 h post-transfection, relative luciferase activity was measured to evaluate ASO-mediated gene silencing efficiency. In all three cell lines, the Luc ASO-treated groups exhibited significantly decreased relative luciferase activity, with knockdown efficiencies ranging from 70% to 84%. These results indicated that the MCE siRNA/miRNA Transfection Reagent (HY-K2017) was suitable not only for siRNA and miRNA delivery but also for efficient delivery of antisense oligonucleotides (ASOs), achieving specific target gene silencing.

MedChemExpress Validation
The siRNA/miRNA Transfection Reagent (HY-K2017) was used to deliver 20 pmol siRNA into seven different cell lines. At 48 h post-transfection, cell viability was assessed via CCK-8 assay, and cell survival rates were calculated relative to untransfected controls (set as 1). In all seven tested cell lines, cell viability remained above 90% after transfection. The optimized formulation of this transfection reagent enabled efficient gene silencing with minimal cytotoxicity, preserving normal cellular physiological status and ensuring more accurate and reliable experimental results.