Purified bovine ubiquitin conjugated to carrier protein.
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 4C, 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 4C. Immunofluorescence: 2-5ug/ml Optimal dilutions to be determined by the researcher. Storage and Stability: Store product at 4C if to be used immediately within two weeks. For long-term storage, aliquot to avoid repeated freezing and thawing and store at -20C. Aliquots are stable at -20C for 12 months after receipt. Dilute required amount only prior to immediate use. Further dilutions can be made in assay buffer. Caution: FITC conjugates are sensitive to light. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap. Note: Applications are based on unconjugated antibody.
