This is a knockout-validated antibody summary, based on the publication "Distinct SoxB1 networks are required for naïve and primed pluripotency", as cited below [1]. Labome curates formal publications to compile a list of antibodies with unambiguous specificity within Validated Antibody Database (VAD).
Rabbit monoclonal IgG
Company: Abcam
Antibody: SOX2
Catalog number: ab92494
Summary: Rabbit monoclonal IgG against a synthetic peptide within human SOX2. Reacts with mouse, human, Leucoraja erinacea. Suitable for western blot, immunohistochemistry (paraffin and whole mount), immunocytochemistry/immunofluorescence and ELISA. Unsuitable for flow cytometry or immunoprecipitation.
Western blot | Immunocytochemistry
E14Tg2a, Sox2fl/- and Sox2-/- epiblast stem cells (EpiSCs).
WB: Cells were lysed with lysis buffer comprising 50 mM Tris pH 8.0, 150 mM NaCl supplemented with fresh 0.5% NP-40, 0.5 mM DTT, 1 × protease inhibitors cocktail (Roche, Switzerland) and 1.3 μl of Benzonase (Novagen, Germany) (1 hr, 4°C).
IC: Cells were fixed with 4% PFA (10 mins, RT) and permeabilized using PBS/0.1% (v/v) Triton-X100 (PBSTr) for 10 mins, before quenching with 0.5M Glycine/PBSTr (15 mins).
WB: 5% (w/v) low-fat milk in 0.01% (v/v) Tween-20/PBS (PBSTw).
IC: 3% (v/v) donkey serum/1% (v/v) BSA/PBST for 1 hr at RT.
WB: 1:1,000 dilution in blocking solution.
IC: 1:400 dilution in blocking solution at 4°C overnight.
WB: 1:10,000 dilution donkey-raised secondary antibodies conjugated with IRDye 800CW (LI-COR 926–32213, Lincoln, USA).
IC: donkey-raised secondary antibodies conjugated with Alexa-488 in blocking solution for 1–2 hr at RT.
WB: Supersignal West Pico kit (Pierce, Cambridge, USA) and LI-COR Odyssey Fc imager.
IC: Olympus IX51 inverted fluorescent microscope.
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