Application
Western blot
Sample
Human Tissue lysate - whole (whole human acute phase plasma)
Loading amount
14 µg
Specification
whole human acute phase plasma
Treatment
heated to 95C in 10-fold excess of reducing SDS sample buffer for 10min
Gel Running Conditions
Reduced Denaturing (4-12% Bis-Tris)
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 20°C
Other product details
Dilution
1/4000
Incubation time
1 hour(s) and 0 minute(s) · Temperature: 20°C · Diluent: PBS, Tween-20 (0.1%)
Secondary antibody
Name
Non-Abcam antibody was used: Goat anti-mouse IgG-HRP (Dako P0447)
Host species: Goat
Clonality: Polyclonal
Conjugation: Horse Radish Peroxidase
Host species: Goat
Clonality: Polyclonal
Conjugation: Horse Radish Peroxidase
Dilution
1/10000
Detection
Detection method
ECL+
Exposure
5 second(s)
Bands
Specific: 12 kDa Non-specific: none kDa
Positive control
recombinant A-SAA (MBL International, JM-4324)
Negative control
Non-acute phase plasma C-SAA protein (recombinant)
Additional data
PubMed ID
20463814: View
Additional Notes
Used PVDF membrane (Immobilon P, Millipore, #IPVH0010) and Kodak Biomax XAR film (Sigma-Aldrich, # F5513) for detection.
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 21 2011