Technical Data

U1000-04B-PE
Clone Type
Monoclonal
Host
Mouse
Source
Bovine
Conjugate
PE
Isotype
IgG1,k
Clone Number
6A209
Grade
Affinity Purified
Applications
FLISA IF IHC WB
Crossreactivity
Bo Ce Ch Dr Hu Mo
Shipping Temp
Blue Ice
Storage Temp
4°C Do Not Freeze
Notes
Preservative Free
BSA Free
Mouse Anti-Ubiquitin-PE
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: PE 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.
Immunogen
Purified bovine ubiquitin conjugated to carrier protein.
Form
Supplied as a liquid in PBS, pH 7.2. No preservatives added. Labeled with R-Phycoerythrin (PE).
Purity
Purified by Protein A affinity chromatography.
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.

Intended for research use only. Not for use in human, therapeutic, or diagnostic applications.

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
No references available
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