SOX2 antibody | knockout validation | eBioscience 14-9811-80

This is a knockout-validated antibody summary, based on the publication "One-step generation of conditional and reversible gene knockouts", as cited below [1]. Labome curates formal publications to compile a list of antibodies with unambiguous specificity within Validated Antibody Database (VAD).

Antibody information

Rat monoclonal IgG2a, kappa

Company: eBioscience

Antibody: SOX2

Catalog number: 14-9811-80

Summary: Rat monoclonal IgG2a, kappa against SOX2. Reacts with human and mouse. Suitable for western blot, immunocytochemistry/immunofluorescence, immunohistochemistry (paraffin), flow cytometry and ELISA.

Validation Method

Western blot | Immunocytochemistry

Sample

Control and mouse embryonic stem cells (mESCs). Cells were lysed in buffer containing complete protease-inhibitor cocktail tablets (11697498001, Roche).

Blocking agent

IC: 5% donkey serum (D9663, Sigma) and 0.1% Triton X-100 for 1 h at RT.

Primary incubation

WB: 1:500 dilution.

IC: 1:500 dilution in blocking buffer overnight at 4 °C.

Secondary incubation

WB: 1:5,000 dilution goat anti-rat HRP conjugated (SC2032, Santa Cruz).

IC: 1:1,000 dilution anti-rat Alexa Flour 594-conjugated antibody (A21209, Abcam) for 1 h at RT.

Detection

WB: ECL prime western blotting detection system (RPN2133, GE Healthcare).

Figure

Loss of protein following Cre recombination was confirmed by immunofluorescence (h) and western blot (i). Please see Supplementary Figure 6h, i in the article [1].

References
  1. Andersson Rolf A, Mustata R, Merenda A, Kim J, Perera S, Grego T, et al. One-step generation of conditional and reversible gene knockouts. Nat Methods. 2017;14:287-289 pubmed publisher