Application
Western blot
Sample
Human Recombinant protein (di-SUMO recombinant protein in cleavage assay)
Loading amount
0.11 µg
Specification
di-SUMO recombinant protein in cleavage assay
Treatment
50nM of specific protease or 5ug HEK293T cell lysate as a positive control (0.5-15 h)
Gel Running Conditions
Reduced Denaturing (4-13% Bis-Tris)
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Other product details
Dilution
1/1000
Incubation time
1 hour(s) and 0 minute(s) · Temperature: 25°C · Diluent: phosphate buffered saline, 0.1% Tween-20
Secondary antibody
Name
Non-Abcam antibody was used: Goat anti-rabbit IgG-HRP (P0449)
Host species: Goat
Clonality: Polyclonal
Conjugation: Horse Radish Peroxidase
Host species: Goat
Clonality: Polyclonal
Conjugation: Horse Radish Peroxidase
Dilution
1/20000
Detection
Detection method
ECL+
Exposure
30 second(s)
Bands
Specific: 20, 12 kDa Non-specific: none kDa
Positive control
Di-SUMO2 recombinant protein, BostonBiochem (ULC-200)
Negative control
n/a
Additional data
PubMed ID
18954305: View
Additional Notes
Time course (4 time points, hours) of a di-SUMO2 deconjugation assay in order to test specificity of a ubiquitin deconjugating enzyme (protease OTUB2) towards SUMO2-conjugates. Positive cotrol for cleavage is HEK293T lysate.
Di-SUMO2 protein was purchased from BostonBiochem (ULC-200).
Di-SUMO2 protein was purchased from BostonBiochem (ULC-200).
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.
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提交于 Jul 28 2011