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U1000-04B-ML490 Ubiquitin (MaxLight 490)

Specifications
References
Brand
MaxLight™
Conjugate
MaxLight™490
Specificity
Recognizes bovine ubiquitin, both conjugated and unconjugated. Reacts with a single chain 8.5kD protein. The ubiquitin molecule appears to be present in all eukaryotic cells and has an identical primary structure in all animals. Ubiquitin is present in the nucleus, cytoplasm, and on cell surface membranes. Species Crossreactivity: human, chicken, mouse, C. elegans and Drosophila.
Source Antibody
bovine
Purity
Purified by Protein A affinity chromatography.
EU Commodity Code
30021010
Shipping Temp
Blue Ice
Storage Temp
4°C Do Not Freeze
Notes
Preservative Free
BSA Free

MaxLight™ 490 is a new Blue-Green photostable dye conjugate comparable to DyLight™488, Alexa Fluor™488 and offers better labeling efficiency, brighter imaging and increased immunodetection. Absorbance (491nm); Emission (515nm); Extinction Coefficient 73,000.

Applications
Suitable for use in FLISA, Western Blot, Immunohistochemistry and Immunoflourescence. Other applications not tested.
Recommended Dilution
FLISA: 0.1-1ug/ml Western Blot: 1-3ug/ml; Prepare lysis buffer in 10mM N-ethylmaleimide to inhibit ubiquitin-conjugating enzymes. N-ethylmaleimide inactivates certain enzymes by blocking free sulfhydryls. After electrophoresis and transfer, pre-incubate transferred membranes in denaturing buffer (6M guanidine-HCl 20mM Tris-HCl, pH 7.5, 5mM b-mercaptoethanol, 1mM PMSF) for 30-60 minutes at 4°C, followed by extensive PBS washing. Immunohistochemistry: Alcohol/paraformaldhyde-fixed paraffin-embedded or Frozen tissue sections. Requires heat induced epitope retrieval with citrate buffer (pH, 6.0) for formalin-fixed, paraffin-embedded sections. To stain, incubate for 30-60 minutes at RT or overnight at 4°C. Immunofluorescence: 2-5ug/ml Optimal dilutions to be determined by the researcher.
Storage and Stability
Store product at 4°C in the dark. DO NOT FREEZE! Stable at 4°C for 12 months after receipt as an undiluted liquid. Dilute required amount only prior to immediate use. Further dilutions can be made in assay buffer. Caution: MaxLight™490 conjugates are sensitive to light. For maximum recovery of product, centrifuge the original vial prior to removing the cap.
Note: Applications are based on unconjugated antibody.
References
1. Manetto, V., et al., PNAS 85: 4501-4505 (1988). 2. Shaw, G. & Chau, V., PNAS 85: 2854-2858 (1988). 3. Askanas, V., et al., Neurosci. Lett. 130: 73-76 (1991). 4. Schmidt, M.L., et al., Am. J. Pathol. 139: 53-65 (1991). 5. Mayer, R.J., et al., Ann. NY Acad. Sci. 674: 149 (1992). 6. Hirano, S., et al., Cell 70: 293-301 (1992). 7. Sternsdorf, T., et al., J. Cell Biol. 139(7): 1621-1634 (1997). 8. Kim, T-W., et al., J. Biol. Chem. 272: 11,006-11,010 (1997). 9. Cuervo, A.M., et al., Mol. Biol. Cell 9: 1995-2010 (1998). 10. Verdier, F., et al., J. Biol. Chem. 273(43): 28,185-28,190 (1998). 11. Laroia, G., et al., Science 284(5413): 499-502 (1999). 12. Marti, A., et al., Nature Cell Biol. 1(1): 14-19 (1999). 13. Sternsdorf, T., et al., Mol. Cell Biol. 19(7): 5170-5178 (1999). 14. Mullally, J.E., et al., J. Biol. Chem. 276(32):3 0,366-30,373 (2001). 15. Ma, Q. & Baldwin, K.T., J. Biol. Chem. 275(12): 8432-8438 (2000). 16. Fernandez-Funez, P., et al., Nature 408: 101-106 (2000). 17. Holbert, S., et al., PNAS 98(4): 1811-1816 (2001). 18. Ruffner, H., et al., PNAS 98(9): 5134-5139 (2001). 19. Roig, J., et al., PNAS 97(26): 14,346-14,351 (2000).
USBio References
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