Purified bovine ubiquitin conjugated to carrier protein.
Applications: Suitable for use in ELISA, Western Blot, Immunohistochemistry and Immunoflourescence. Other applications not tested. Recommended Dilution: ELISA: 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. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap. Note: Applications are based on unconjugated antibody.
