A human siRNA against FXYD3 (consisting of pools of three to five target-specific 19–25-nt siRNAs designed to knock down gene expression, sc-60665) and the non-silencing control (control siRNA-A, sc-37007) were purchased from Santa Cruz Biotechnology (Dallas, TX, USA). Cells were transfected using the siRNA Transfection Reagent/Medium (Santa Cruz Biotechnology) according to the manufacturer’s instructions. The FXYD3 mRNA expression level was quantified by real-time polymerase chain reaction (RT-PCR) as described previously (9 (link)). FXYD3 protein abundance was measured by Western blotting (9 (link)).
Sc 60665
Manufactured by Santa Cruz Biotechnology
Sourced in United States
Sc-60665 is a laboratory instrument designed for the purification and analysis of biomolecules. It utilizes centrifugal force to separate and concentrate samples based on their density and sedimentation properties. The core function of this equipment is to facilitate the isolation and purification of various biological materials, such as proteins, nucleic acids, and cellular components, from complex mixtures.
Automatically generated - may contain errors
2 protocols using sc 60665
1
FXYD3 Gene Silencing Protocol
2
Investigating FXYD3 Knockdown Effects
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
A human siRNA against FXYD3 (consisting of pools of three to ve target-speci c 19-25nt siRNAs designed to knockdown gene expression, sc-60665), and the non-silencing control (control siRNA-A, sc-37007) were purchased from Santa Cruz Biotechnology. Cells were transfected using the siRNA Transfection Reagent/Medium (Santa Cruz Biotechnology) according to the manufacturer's instructions. FXYD3 mRNA expression level was quanti ed by real-time RT-PCR after exposure to FXYD3 siRNA as described previously [5] (link). FXYD3 protein abundance was measured by western blotting using GADPH as a loading control [5] (link).
Cell viability and apoptosis. Cell metabolic activity was assayed as described [13] (link) by estimating reduction of XTT (2,3-bis(2-methoxy-4-nitro-5-sulfophenyl)-2H-tetrazolium-5 carboxyanilide), using a kit (Cell Signaling Technology). The metabolic activity was used as a surrogate for viability. For each set of experimental groups, we performed 5 experiments with 4 replicates. Caspase-3-like activity is increased through a protease cascade during the early stage of apoptosis [14] (link) and we measured activities of Caspase 3/7 (DEVDase) using the Caspase uorogenic substrate (Calbiochem) as described [5] (link).
Cell viability and apoptosis. Cell metabolic activity was assayed as described [13] (link) by estimating reduction of XTT (2,3-bis(2-methoxy-4-nitro-5-sulfophenyl)-2H-tetrazolium-5 carboxyanilide), using a kit (Cell Signaling Technology). The metabolic activity was used as a surrogate for viability. For each set of experimental groups, we performed 5 experiments with 4 replicates. Caspase-3-like activity is increased through a protease cascade during the early stage of apoptosis [14] (link) and we measured activities of Caspase 3/7 (DEVDase) using the Caspase uorogenic substrate (Calbiochem) as described [5] (link).
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!