Application
Western blot
Sample
Mouse Tissue lysate - other (brain)
Loading amount
15 µg
Specification
brain
Gel Running Conditions
Reduced Denaturing
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Other product details
Concentration
1 µg/ml
Incubation time
14 hour(s) and 0 minute(s) · Temperature: 4°C
Secondary antibody
Name
Non-Abcam antibody was used: anti - rabbit
Host species: Goat
Clonality: Polyclonal
Conjugation: Horse Radish Peroxidase
Host species: Goat
Clonality: Polyclonal
Conjugation: Horse Radish Peroxidase
Dilution
1/20000
Detection
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 data
Additional Notes
Blocking peptide is used. Nothing is blocked. Peptide usage 5 times - 1ug of antibody and 5ug of blocking peptide.
Have you tried reducing conditions?
I have tried Reducing condition only, by mistake i chosen non - reducing condition
Would it be possible to send image of your results?
Yes, I will upload the image as soon as possible
Have you tried any optimization of your protocol?
I have tried different protein and antibody concentration
Could you confirm that the secondary antibody was used at 1/20 000 dilution?
Yes, secondary antibody dilution is 1: 20,000
Is it possible to try human samples like Jurkat cells?
sorry, I am not working in human sample
Have you tried reducing conditions?
I have tried Reducing condition only, by mistake i chosen non - reducing condition
Would it be possible to send image of your results?
Yes, I will upload the image as soon as possible
Have you tried any optimization of your protocol?
I have tried different protein and antibody concentration
Could you confirm that the secondary antibody was used at 1/20 000 dilution?
Yes, secondary antibody dilution is 1: 20,000
Is it possible to try human samples like Jurkat cells?
sorry, I am not working in human sample
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.
We have not had similar complaints for Aromatase peptide (ab51924).
More blocking peptide might be needed in customer's case.
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.
Abcam user community
Verified customer
提交于 Apr 28 2011