This is a knockout-validated antibody summary, based on the publication "Giantin-knockout models reveal a feedback loop between Golgi function and glycosyltransferase expression", as cited below [1]. Labome curates formal publications to compile a list of antibodies with unambiguous specificity within Validated Antibody Database (VAD).

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Figure 1. Western blot analysis of giantin in WT and giantin KO RPE-1 cell lines using antibodies aginst full-length (FL) protein. All immunoreactivity is lost in the KO cells. From [1].

Mouse monoclonal IgG1

Company: Abcam

Antibody: Giantin

Catalog number: ab37266

Summary: Mouse monoclonal IgG1 against full length protein of rat Giantin. Reacts with rat and human. Suitable for immunocytochemistry/immunofluorescence and immunohistochemistry (paraffin).

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Figure 2. Immunofluorescence (IF) staining of giantin in WT and giantin KO RPE-1 cell lines using antibodies aginst full-length (FL) protein. Antibodies against the C-terminus (C-term) were also used in the IF. All immunoreactivity is lost in the KO cells. From [1].

Western blot | Immunocytochemistry

WB: Cells were lysed in RIPA buffer (50 mM Tris-HCl, pH 7.5, 300 mM NaCl, 2% Triton X-100, 1% deoxycholate, 0.1% SDS, 1 mM EDTA).

WB: 5% milk in TBST (TBS with 0.05% Tween).

IC: 3% BSA-PBS for 30 min

WB: overnight.

IC: 1:500 dilution for 1 h.

WB: HRP-conjugated secondary antibodies (Jackson ImmunoResearch).

IC: for 1h.

WB: enhanced chemiluminescence (GE Healthcare).

IC: Olympus IX70 microscope with 60×1.42 NA oil-immersion lens, Exfo 120 metal halide illumination with excitation, dichroic and emission filters (Semrock, Rochester, NY), and a Photometrics Coolsnap HQ2 CCD, controlled by Volocity 5.4.1 (Perkin Elmer).