Western Blot: Mouse TLR13 NF-kB/SEAP - (SEAPorter™) Stable Reporter Cell Line [NBP2-26290] - Analysis of TLR13 expression in the TLR13 reporter cell line using an HA antibody (20 ug total protein/lane). Legend. Cont: Control, NF-kB/SEAPorterTM HEK 293 cell line; TLR13: TLR13 reporter cell line.
Flow Cytometry: Mouse TLR13 NF-kB/SEAP - (SEAPorter™) Stable Reporter Cell Line [NBP2-26290] - Expression of TLR13 in the TLR13 reporter cell line was analyzed by flow cytometry using a combination of TLR13 antibody and APC-conjugated Goat Anti-Rabbit IgG (F(ab')2-specific). This cell line was compared with the NF-kB/SEAPorter HEK 293 cell line. Flow samples were prepared using IMGENEX's Intracellular TLR Staining Flow Kit.
Ligand Activation: Mouse TLR13 NF-kB/SEAP - (SEAPorter™) Stable Reporter Cell Line [NBP2-26290] - TLR13 reporter cells were plated in a 96-well plate at 5 x 10^4 cells per well. Cells were stimulated with various concentrations of phorbol 12-myristate 13-acetate (PMA), which is a positive control for NF-kB induction, for 24 h. SEAP was analyzed using the SEAPorter Assay Kit. Data Summary: PMA is a positive control for NF-kB induction. PMA stimulation confirmed that NF-kB/SEAP reporter is functional in TLR13 SEAP.
Ligand Activation: Mouse TLR13 NF-kB/SEAP - (SEAPorter™) Stable Reporter Cell Line [NBP2-26290] - TLR13/NF-kB SEAPorter (TM) HEK 293 cells were plated in a 96-well plate at 5 x 10^4 cells per well. Cells were stimulated with various amounts of total bacterial RNA packed with Lipofectamine 2000 (A), phenol extracted total bacterial RNA packed with Lipofectamine 2000 (B), bacterial total RNA alone (C) and poly(I:C) packed with Lipofectamine 2000 (D) for 24 h. SEAP was then analyzed using the SEAPorter Assay Kit. Data Summary: TLR13 in the TLR13/NF-kB SEAPorter (TM) HEK 293 cell line was only activated by the bacterial RNA packed with Lipofectamine 2000 (A & B) but not by the unpacked bacterial RNA (C) or poly(I:C) packed with Lipofectamine 2000 (D).