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Recognizes the ~34 kDa Sox2 protein in NTERA-2 cells.
Catalogue Number
SC1002
Brand Family
Calbiochem®
References
References
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.
Product Information
Form
Lyophilized
Formulation
Lyophilized from 0.2 µm-filter sterilized PBS, 5% trehalose.
Immunoblotting (1-2 µg/ml) Immunocytochemistry (10 µg/ml; see comments) Flow Cytometry (see comments)
Application Comments
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.
Biological Information
Immunogen
a recombinant protein consisting of amino acids 135-317 of human Sox2, expressed in E. coli
Immunogen
Human
Clone
245610
Host
Mouse
Isotype
IgG2a
Species Reactivity
Human
Mouse
Antibody Type
Monoclonal Antibody
Physicochemical Information
Dimensions
Materials Information
Toxicological Information
Safety Information according to GHS
Safety Information
Product Usage Statements
Storage and Shipping Information
Ship Code
Blue Ice Only
Toxicity
Standard Handling
Storage
-20°C
Do not freeze
Ok to freeze
Special Instructions
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.
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.
数据表
Note that this data sheet is not lot-specific and is representative of the current specifications for this product. Please consult the vial label and the certificate of analysis for information on specific lots. Also note that shipping conditions may differ from storage conditions.
Revision
17-January-2008 JSW
Application
Immunoblotting (1-2 µg/ml) Immunocytochemistry (10 µg/ml; see comments) Flow Cytometry (see comments)
Description
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.
Background
Sox2 is a member of the Sox (SRY-like HMG box) family of transcription factors, which exhibit diverse roles in development. Sox2 plays a role in specifying the first three lineages that are present at egg implantation. In addition, Sox2 functions in regulating proliferation and differentation in the developing peripheral nervous system. As a result of its role in development, Sox2, together with Oct4 and Nanog, is also essential for the propagation of undifferentiated stem cells.
Host
Mouse
Immunogen species
Human
Immunogen
a recombinant protein consisting of amino acids 135-317 of human Sox2, expressed in E. coli
Clone
245610
Isotype
IgG2a
Species
human, mouse
Positive control
NTERA-2 cells
Form
Lyophilized
Formulation
Lyophilized from 0.2 µm-filter sterilized PBS, 5% trehalose.
Comments
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.
Storage
-20°C
Do Not Freeze
Ok to freeze
Special Instructions
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.
Toxicity
Standard Handling
References
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.
