IHC-plus(tm) Ubiquitin Antibody

LifeSpan BioSciences

IHC-plus(tm) Ubiquitin Antibody

23

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. Species cross-reactivity:

Immunohistochemistry: LS-B3328 was validated for use in immunohistochemistry on a panel of 21 formalin-fixed, paraffin-embedded (FFPE) human tissues after heat induced antigen retrieval in pH 6.0 citrate buffer. After incubation with the primary antibody, slides were incubated with biotinylated secondary antibody, followed by alkaline phosphatase-streptavidin and chromogen. The stained slides were evaluated by a pathologist to confirm staining specificity. The optimal working concentration for LS-B3328 was determined to be 10 ug/ml. Western Blot: 1-3 ug/ml; Prepare lysis buffer in 10 mM 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 20 mM Tris-HCl, pH 7.5, 5 mM b-mercaptoethanol, 1 mM PMSF) for 30-60 minutes at 4uC, followed by extensive PBS washing.. ELISA (0.1 - 1 µg/ml), IF, IHC, IHC-Fr, IHC-P (10 µg/ml), WB Immunohistochemistry: LS-B3328 was validated for use in immunohistochemistry on a panel of 21 formalin-fixed, paraffin-embedded (FFPE) human tissues after heat induced antigen retrieval in pH 6.0 citrate buffer. After incubation with the primary antibody, slides were incubated with biotinylated secondary antibody, followed by alkaline phosphatase-streptavidin and chromogen. The stained slides were evaluated by a pathologist to confirm staining specificity. The optimal working concentration for LS-B3328 was determined to be 10 ug/ml. Western Blot: 1-3 ug/ml; Prepare lysis buffer in 10 mM 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 20 mM Tris-HCl, pH 7.5, 5 mM b-mercaptoethanol, 1 mM PMSF) for 30-60 minutes at 4uC, followed by extensive PBS washing.

ImmunoFluorescence, Western Blot, ELISA, ImmunoHistoChemistry, ImmunoHistoChemistry Frozen, ImmunoHistoChemistry Paraffin

Research Use Only

Monoclonal

0.5 mg/ml

Unconjugated

...

Mouse

IgG1

Bovine, Human, Mouse

-20°C,2°C to 8°C

12352203