Application
Western blot
Sample
Human Cell lysate - whole cell (U2OS and 293T cell line)
Loading amount
30 µg
Specification
U2OS and 293T cell line
Gel Running Conditions
Reduced Denaturing
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 21°C
Other product details
Concentration
1 µg/ml
Incubation time
12 hour(s) and 0 minute(s) · Temperature: 4°C · Diluent: 5% Milk 0.1% Tween-20 in PBS
Secondary antibody
Name
Non-Abcam antibody was used: Goat anti-mouse HRP
Host species: Goat
Clonality: Polyclonal
Conjugation: Horse Radish Peroxidase
Host species: Goat
Clonality: Polyclonal
Conjugation: Horse Radish Peroxidase
Dilution
1/5000
Detection
Detection method
Supersignal West Dura
Exposure
4 minute(s) and 0 second(s)
Bands
Specific: 60 kDa Non-specific: 40 (in several cell kDa
Additional data
Additional Notes
I did not see a species of the predicted mobility (72 kDa) in any of the 6 cell lines tested. Note: Cox-1 is constitutively expressed in nearly all tissues/ cell lines, including PC12 and 3T3 - which are known to express immunodetectable levels of Cox-1, and is consistent with the 60 kDa band being correctly assigned.
I have given this antibody 5 stars on the basis of the ease of use/ signal strength. Further work, e.g., using siRNA to Cox-1, would be helpful in formally assigning the 60 kDa band as Cox-1
Lane Order
1: U2OS (Hu) 2: 293T (Hu) 3: Cos-7 (AGMk) 4: CHO (Hm) 5: PC12 (Rat) 6: 3T3 (Ms)
I have given this antibody 5 stars on the basis of the ease of use/ signal strength. Further work, e.g., using siRNA to Cox-1, would be helpful in formally assigning the 60 kDa band as Cox-1
Lane Order
1: U2OS (Hu) 2: 293T (Hu) 3: Cos-7 (AGMk) 4: CHO (Hm) 5: PC12 (Rat) 6: 3T3 (Ms)
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 Nov 07 2007