Technical Tips: Gel Shift Assays
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1. Can Oncogene Research Products' antibodies be used for gel shift assays? Top
Many people have used our antibodies successfully in gel shift assays (1-11). References:
1. Chiles, T.C. et al. (1991) J. Immunol. 146: 1730-1735
2. Jain, J. (1992) Nature 356: 801-804
3. Hughes, M. et al. (1992) Cell Growth Differ. 3: 889-897
4. Kastan, M.B. et al. (1992) Cell 71: 587-597
5. Hupp, T.R. et al. (1992) Cell 71: 875-886
6. Halazonetis, T.D. (1993) EMBO J. 12: 1021-1028
7. Zhang, W. et al. (1993) Oncogene 8: 2555-2559
8. Srivastava, S. et al. (1993) Cancer Res. 53: 4452-4455
9. Wu, Y. et al. (1994) EMBO J. 13: 4823-4830
10. Waterman, J. et al. (1995) EMBO J. 14: 512-519
11. Benson, B.A. et al. (1994) J. Immunol. 153: 3862-3873
2. How do I use Oncogene Research Products' in gel shift? Top
Use 1 µg of antibody in 1 µl per sample. The antibody stock solution should be 1 mg/ml; purchase lyophilized antibodies (the catalog number will end in an "L") and bring them up in 100 µl (rather than 1 ml as we usually recommend). If the antibody you need does not come standardly in a lyophilized form please inquire as to avaiability. There is usually a fee for special vialings. Please note that our antibodies sold in solution are too dilute (100 µg/ml) for use in gel shift assays.
3. Which antibodies gel shift? Top
Antibodies for which we have citations for use in gel shift assays include:
jun (Ab-1), Cat# PC06L (1-3); blocks the gel shift if it is bound before DNA is added
jun (Ab-2), Cat# PC07L (2); supershifts
p53 (Ab-1), Cat# OP03L (4-10)
p53 (Ab-2), Cat# OP09L (4)
p53 (Ab-3), Cat# OP29L (5)
p53 (Ab-5), Cat# OP33L (5)
p53 (Ab-6), Cat# OP43L (5)
phospho-tyrosine (Ab-1), Cat# PT01L (11)
References:
Chiles, T.C. et al. (1991) J. Immunol. 146: 1730-1735
Jain, J. (1992) Nature 356: 801-804
Hughes, M. et al. (1992) Cell Growth Differ. 3: 889-897
Kastan, M.B. et al. (1992) Cell 71: 587-597
Hupp, T.R. et al. (1992) Cell 71: 875-886
Halazonetis, T.D. (1993) EMBO J. 12: 1021-1028
Zhang, W. et al. (1993) Oncogene 8: 2555-2559
Srivastava, S. et al. (1993) Cancer Res. 53: 4452-4455
Wu, Y. et al. (1994) EMBO J. 13: 4823-4830
Waterman, J. et al. (1995) EMBO J. 14: 512-519
Benson, B.A. et al. (1994) J. Immunol. 153: 3862-3873
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