Suitable for use in ELISA, Western Blot, Immunohistochemistry and Immunoflourescence. Other applications not tested.

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 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.

May be stored at 4°C for short-term only. Aliquot to avoid repeated freezing and thawing. Store at -20°C. Aliquots are stable for 12 months. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap.

Purified bovine ubiquitin conjugated to carrier protein.

Supplied as a liquid in PBS, pH 7.4, 0.09% sodium azide.

Purified by Protein A affinity chromatography.

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.