Anti-Sox2 Mouse mAb (245610)

Analysis Note

Positive ControlNTERA-2 cells

Application

Immunoblotting (1-2 µg/ml)r>Immunocytochemistry (10 µg/ml; see comments)r>Flow Cytometry (see comments)

General description

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.

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.

Immunogen

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

Human

Legal Information

Triton is a trademark of The Dow Chemical Company or an affiliated company of Dow

CALBIOCHEM is a registered trademark of Merck KGaA, Darmstadt, Germany

Other Notes

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.

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.

Packaging

100 µg in Plastic ampoule

Physical form

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

Reconstitution

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.

Warning

Toxicity: Standard Handling (A)