Fluorescent TrueBlot^(R): Anti-IgG  

Immunofluorescence microscopy of ZO-1 in Caco-2 cells using FITC-conjugated Fluorescent TrueBlotR anti-rabbit IgG for detection. Caco-2 cells were fixed with 4% PFA, blocked (5% mouse serum/0.3% Triton X-100 in 1X PBS) for 1 hr, then incubated with 15 μg/mL of anti-ZO-1 primary antibody (Cat. No. 600-401-GU7) at 4°C overnight. Following 3 washes in 1X PBS for 5 min each, 5 μg/mL of FITC-conjugated Fluorescent TrueBlotR anti-rabbit IgG was added and allowed to incubate for 1 hr at room temperature. Nuclei were counterstained with DAPI present in mounting medium. Predicted cell localization is cell membrane and cell junctions. Image taken at 40X magnification. (right) Merged DAPI (blue)/ZO-1 (green), image shown (left) secondary antibody only.

Immunofluorescence microscopy of BCL3 in Caco-2 cells using FITC-conjugated Fluorescent TrueBlotR anti-rabbit IgG for detection. Caco-2 cells were fixed with 4% PFA, blocked (5% mouse serum/0.3% Triton X-100 in 1X PBS ) for 1 hr, then incubated with 15 μg/mL of anti-BCL3 primary antibody (Cat. No. 600-401-GU4) at 4°C overnight. Following 3 washes in 1X PBS for 5 min each, 5 μg/mL of FITC-conjugated Fluorescent TrueBlotR anti-rabbit IgG was added and allowed to incubate for 1 hr at room temperature. Nuclei were counterstained with DAPI present in mounting medium. The predicted main localization is nucleoplasm. Additional localization in some cell types includes vesicles and midbody. (a) BCL3 (b) DAPI (c) merged DAPI/BCL3 (d) secondary antibody only. Image taken at 40X magnification.

Rabbit TrueBlotR IP / Western Blot: Jurkat cell lysate (0.5 ml of 1x10e7 cells/ml) was incubated with rabbit anti-human Stat1 and immunoprecipitated using Protein G, Protein A and Anti-Rabbit Ig IP Beads. Precipitate from 5x10e5 cells was subjected to electrophoresis, transferred to a PVDF membrane, and Western blotted with anti-Stat1 using Rabbit TrueBlotR: Anti-Rabbit IgG HRP

Western Blot of Fluorescent TrueBlotR: Anti-Rabbit IgG Fluorescein. Lane 1: Rabbit IgG, Non-reduced. Lane 2: Rabbit IgG, Reduced. Load: 50 ng per lane. Primary antibody: none. Secondary antibody: Fluorescent TrueBlotR: Anti-Rabbit IgG Fluorescein at 1:1,000 for 60 min at RT. Block: MB-070 for 30 min at RT. Predicted/Observed size: 160 kDa for Rabbit IgG, Non-reduced. Other band(s): none.