Application
Western blot
Sample
Human Cell lysate - whole cell (THP-1 human leukemic monocyte)
Gel Running Conditions
Reduced Denaturing (12%)
Loading amount
20 µg
Treatment
100 nM PMA for 24 hrs, 100 ng/ml LPS for 4 hrs
Specification
THP-1 human leukemic monocyte
Blocking step
Odyssey Blocking Buffer as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 50% · Temperature: 22°C
Other product details
Incubation time
16 hour(s) and 0 minute(s) · Temperature: 4°C
Dilution
1/1000
Secondary antibody
Dilution
1/5000
Name
Non-Abcam antibody was used: Donkey anti-Rabbit LI-COR
Host species: Donkey
Clonality: Monoclonal
Conjugation: IRDye® 800CW
Host species: Donkey
Clonality: Monoclonal
Conjugation: IRDye® 800CW
Detection
Detection method
IR imaging (Odyssey)
Negative control
no mature IL-1beta band(p17), only pro-IL-1beta band (p31) was detected in supernatant from THP-1 cells not stimulated with LPS
Exposure
30 second(s)
Bands
Specific: 17, 31 kDa
Positive control
both mature and pro-IL-1beta bands, p17 and p31, detected by the ab2105 antibody in supernatant from THP-1 cells stimulated with LPS
Additional data
Additional Notes
Lane Number:
1) ladder (top to bottom: p35, p25, p20, p17, p11)
2) THP-1 cell lysate, no LPS (p31 band)
3) THP-1 supernatant, no LPS (p31 band)
4) THP-1 cell lysate, LPS stimulation (p31 band)
5) THP-1 supernatant, LPS stimulation (p17 and p31 band)
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投稿 Aug 12 2013