Silencing of Fas in activated TCRP1A CD8+ T cells was achieved by using Accell siRNA (1 µM) (Dharmacon), with sequence 5ʹ(P)-GUGCAAGUGCAAACCAGAC(dTdT)-3ʹ. After 1 day of activation, TCRP1A CD8+ T cells were incubated with 1 µM Accell control or Fas siRNA in IMDM complete medium without serum for 24 h, then the medium was replaced with IMDM complete medium with serum. The silencing procedure was repeated on day 3. On day 4, 107 TCRP1A CD8+ T cells were adoptively transferred into mice bearing induced tumors. Silencing efficiency was monitored by Western blot using anti-Fas antibody (Santa Cruz, FL- 335, 1:1000). Blots were stripped and reprobed with a rabbit anti-mouse monoclonal antibody to GAPDH (Cell Signaling, 14C10, 1:5000).
Anti fas antibody
Manufactured by Santa Cruz Biotechnology
The Anti-Fas antibody is a laboratory reagent used for the detection and analysis of the Fas receptor, also known as CD95 or Apo-1. It binds specifically to the Fas receptor, which plays a role in the regulation of programmed cell death or apoptosis. The antibody can be utilized in various immunological techniques, such as flow cytometry, immunohistochemistry, and Western blotting, to study the expression and function of the Fas receptor in different biological systems.
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2 protocols using anti fas antibody
1
Silencing Fas in Activated TCRP1A CD8+ T Cells
2
Immunoprecipitation of Glutathionylated Proteins
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Tissue lysates were prepared from snap-frozen liver tissues according to the manufacturer's instructions (KeyGEN BioTECH). The protein concentration was determined by using the BCA method (P0010, Beyotime) and equalized for the immunoprecipitation assay [22] . The tissue lysate was then incubated with anti-glutathione antibody (Virogen) (2 µg/ml) overnight at 4 °C and immunoprecipitated by protein G agarose beads. The isolated immunoprecipitated proteins were then incubated with the anti-Fas antibody (Santa Cruz). For the control, dithiothreitol (DTT, 50 mM) was added to reduce S-glutathionylated proteins.
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