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Sc 2710281

Manufactured by Santa Cruz Biotechnology
Sourced in United States

Sc-2710281 is a laboratory equipment product. It is designed for use in scientific research and analysis. The core function of this product is to [DESCRIPTION_NOT_AVAILABLE].

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2 protocols using sc 2710281

1

Western Blot Analysis of Inflammatory Markers

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Cell homogenates were studied by Standard Western blot techniques. Briefly, 25 μg of homogenate was separated on a 10% acrylamide gel by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and transferred to polyvinylidene difluoride membranes (Hybond-P, Amersham Pharmacia Biotech, Piscataway, NJ) with a semidry transfer apparatus. The membranes were blocked with Tris Buffered Saline-5% milk and then probed with anti caspase-3, anti TNF-α, anti-IFN-γ and anti- IL-2 (sc-2710281, sc-1350, sc-59993 and sc-48545; 1:200, 1:100, 1:100 and 1:100, respectively, Santa Cruz Biotechnology, Santa Cruz, CA, USA). The primary antibody was detected with horseradish peroxidase-conjugated anti-mouse secondary antibody (sc-2005, diluted 1:5000, 1:2000, 1:1000 and 1:2000 for caspase-3, TNF-α, IFN-γ and IL-2, respectively, Santa Cruz Biotechnology, Santa Cruz, CA, USA). The membranes were washed, and blots were developed using an enhanced chemiluminescence Western blotting detection kit(ECL Plus kit, Amersham Biosciences RPN2132, USA) and exposed to X-ray film (Sigma C4729-1EA, Sigma Z363006-50) for 2–30 s. Molecular weight standards (See Blue® Plus2, LC 5925, Life Technologies, USA) were used to determine molecular weights of the visualized bands.
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2

Quantifying Apoptosis Signaling Proteins

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To find out whether changes in caspase-3 activity and concentrations of TNF-α and IFN-γ were due to changes in protein content, cell homogenates were studied by standard western blot techniques. Briefly, 25 μg of homogenate protein were separated on a 10% acrylamide gel by SDS-PAGE and then transferred to poly vinylidene difluoride membranes (Hybond-P, Amersham Pharmacia Biotech, Piscataway, NJ, USA) with a semidry transfer apparatus. The membranes were blocked with TBS-5% milk and then probed with anti caspase-3, anti -TNF-α, and anti-IFN-γ (sc-2710281, sc-1350, and sc-59993; 1:200, 1:100 and 1:100, respectively, Santa Cruz Biotechnology, Santa Cruz, CA, USA). The primary antibody was detected with horseradish peroxidase-conjugated antimouse secondary antibody (sc-2005, diluted 1:5000, 1:2000 and 1:1000 for caspase-3, TNF-α and IFN-γ, respectively, Santa Cruz Biotechnology, Santa Cruz, CA, USA). The membranes were washed and blots were developed using an enhanced chemiluminescence western blotting detection kit (ECL Plus kit, Amersham Biosciences RPN2132, USA) and exposed to x-ray film (Sigma C4729-1EA, Sigma Z363006-50) for 2-30 s. Molecular weight standarts (See Blue® Plus2, LC 5925, Life Technologies, USA) were used to determine molecular weights of the visualized bands.
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