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 in the dark. DO NOT FREEZE! Stable at 4C 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: APC 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.
