Application
Western blot
Sample
Human Cell lysate - whole cell (HEK293 and hTERT-RPE1)
Gel Running Conditions
Reduced Denaturing (4-12% Bis-Tris, MES buffer)
Loading amount
10 µg
Specification
HEK293 and hTERT-RPE1
Blocking step
BSA as blocking agent for 30 minute(s) · Concentration: 3% · Temperature: 22°C
Other product details
Incubation time
16 hour(s) and 0 minute(s) · Temperature: 4°C · Diluent: 3% BSA in PBS with 0.1% Tween 20
Dilution
1/250
Secondary antibody
Name
Non-Abcam antibody was used: ECL donkey anti-rabbit secondary
Host species: Donkey
Clonality: Monoclonal
Conjugation: Horse Radish Peroxidase
Host species: Donkey
Clonality: Monoclonal
Conjugation: Horse Radish Peroxidase
Detection
Detection method
ECL
Negative control
Whole cell lysates of HEK293 depleted of HOIP
Exposure
5 minute(s) and 0 second(s)
Bands
Specific: over 98 and 60 kDa Non-specific: 25 and others kDa
Positive control
HEK293 and hTERT-RPE1 whole cell lysates.
Additional data
Additional Notes
Samples were transferred by iBlot to a nitrocellulose membrane.
The antibody is very sensitive and detects very well the full length HOIP (over 98 kDa). Also, it detects the short isoform (band around 60 kDa).
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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投稿 Mar 28 2014