Application
Western blot
Sample
Mouse Cell lysate - whole cell (Neuroblasome cells)
Gel Running Conditions
Non-reduced Non-Denaturing (Native)
Loading amount
100 µg
Specification
Neuroblasome cells
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 20°C
Other product details
Incubation time
20 hour(s) and 0 minute(s) · Temperature: 4°C · Diluent: BSA 5% in TBST
Dilution
1/2000
Secondary antibody
Dilution
1/2000
Name
Non-Abcam antibody was used: PrP (FL-253): sc-15312
Host species: Rabbit
Clonality: Polyclonal
Conjugation: Horse Radish Peroxidase
Host species: Rabbit
Clonality: Polyclonal
Conjugation: Horse Radish Peroxidase
Detection
Detection method
ECL
Exposure
1 hour(s), 0 minute(s) and 0 second(s)
Positive control
Lysate from N2a cells
Additional data
Additional Notes
In my experiment I tried to optimize the protocol with this antibody but without any results. Actually I tried to use different concentration of primary from 2500 to 1000 as well as for the secondary.
In my western blotting I'm using RIPA to lysate the cells, Mini-Protean Precast Gels to run the samples, PVDF membrane for the transfer.
Abcam response
Thank you for submitting this review. Your protocol looks good and I believe you should have seen the band around 30 kD given that your samples should express PrP and we have tested the application and species. There may have been a problem with the vial you received. I would be happy to provide a replacement, credit or refund for the product as per our Abpromise guarantee, provided the order was in the last 12 months.
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.
Imane Ghafir El Idrissi
Verified customer
投稿 Feb 12 2016