Review text: How long did you incubate the protein and antibody mix before adding it to the blot?
First Time: Incubation time of antibody and protein (peptide): two hour at room temperature
Second Time:Incubation time of antibody and protein (peptide): One hour at 37 degree C
How much antibody/protein mix was added to the blot?
Antibody used for the reaction is 1ug and protein (peptide) 5ug (1:5 ratio)
Sample: Mouse Tissue lysate - other (BRAIN)
Loading amount: 15 µg
Blocking step: Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Other product details
Concentration: 5 µg/ml
Incubation time: 1 hour(s) and 0 minute(s) · Temperature: 30°C
Primary antibody (in addition to 'Aromatase peptide')
Concentration: 1 µg/ml
Name: Non-Abcam antibody was used: anti - rabbit
Host species: Goat
Conjugation: Horse Radish Peroxidase
Detection method: ECL
Exposure: 2 minute(s) and 0 second(s)
Bands: Specific: 54 kDa Non-specific: 104, 77, 71, 67, 46, 36, kDa
Additional Notes: AROMATASE PEPTIDE DON'T BLOCK ANY BANDS.
Peptide usage 5 times - 1ug of antibody and 5ug of blocking peptide.
Abcam response: The blocking peptide was the same as the immunogen. At the moment we have not found a reason why blocking is not taking place,
We have not had similar complaints for Aromatase peptide (ab51924).
More blocking peptide might be needed in customer's case.
Abcam user community
Submitted Apr 28 2011