Anti-Sox2 Mouse mAb (245610)

Protein G purified mouse monoclonal antibody derived by immunizing mice with the specified immunogen and fusing splenocytes with mouse myeloma cells. Recognizes the ~34 kDa Sox2 protein.

Recognizes the ~34 kDa Sox2 protein in NTERA-2 cells.

This Anti-Sox2 Mouse mAb (245610) is validated for use in Immunoblotting, Immunocytochemistry, Flow Cytometry for the detection of Sox2.

Human

a recombinant protein consisting of amino acids 135-317 of human Sox2, expressed in E. coli

Immunoblotting (1-2 µg/ml)

Immunocytochemistry (10 µg/ml; see comments)

Flow Cytometry (see comments)

Toxicity: Standard Handling (A)

Lyophilized from 0.2 µm-filter sterilized PBS, 5% trehalose.

Reconstitute with 200 µl sterile PBS for a final stock concentration of 500 µg/ml. Following reconstitution, aliquot and freeze (-20°C or -70°C) for long-term storage. Avoid freeze/thaw cycles of solutions. Thawed aliquots are stable for up to 1 month at 4°C.

Positive Control
NTERA-2 cells

Avilion, A.A., et al. 2003.Genes Dev.17, 126.
Graham, V., et al. 2003.Neuron39, 749.
Stevanovic, M. 2003.Mol. Biol. Rep.30, 127.
Kishi, M., et al. 2000.Development127, 791.
Uwanogho, D., et al. 1995.Mech. Dev.49, 23.
Yuan, H., et al. 1995.Genes Dev.9, 2635.

For immunocytochemistry, cells should be fixed in PBS containing 4% paraformaldehyde for 20 min, followed by blocking in PBS/10% normal donkey serum/1% BSA/0.1% Triton^™ X-100 detergent for 45 min and staining overnight at 4°C with appropriately diluted primary antibody. For immunoblotting, the best results may be obtained using whole cell or nuclear extracts; Sox2 may be difficult to detect in cytoplasmic extracts. The detection limit for recombinant Sox2 is ~5 ng/lane under reducing and non-reducing conditions. For intracellular staining by flow cytometry, fix cells in 4% paraformaldehyde and permeabilize with 0.1% saponin. Following fixation and permeabilization, dilute the antibody to 50 µg/ml and add 10 µl diluted antibody to 1-2.5 x 10⁵ cells in a total volume of ≤200 µl. Following incubation with appropriate detection antibody, the cells should be washed a final time with 0.1% saponin prior to analysis. Antibody should be titrated for optimal results in individual systems.

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