Application
Western blot
Sample
Human Cell lysate - whole cell (Breast epithelial cancer cell lines)
Specification
Breast epithelial cancer cell lines
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5%
Other product details
Dilution
1/2000
Incubation time
2 hour(s) and 0 minute(s)
Secondary antibody
Name
Non-Abcam antibody was used: Amersham biosciences ECL Anti-Mouse IgG
Host species: Goat
Clonality: Monoclonal
Conjugation: Horse Radish Peroxidase
Host species: Goat
Clonality: Monoclonal
Conjugation: Horse Radish Peroxidase
Detection
Detection method
ECL
Exposure
1 minute(s) and 0 second(s)
Bands
Specific: 70 kDa
Additional data
Additional Notes
MCF10A were transfected with either (1) control, or PABP-specific (2, 3), siRNAs. The cells were lysed with NP40 buffer with protease inhibitor cocktail, 10 micrograms of protein were separated by SDS-PAGE and transferred to PVDF membrane, blocked and blotted for two hours with PABP antibody at 1:2000 dilution in TBST, washed three times, secondary antibody for 1 hr (goat anti-mouse, Amersham, 1:10000). Only one band at 70 kDa is observed with the anti-PABP antibody.
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.
Ms. Michelle Badura
Verified customer
Submitted Sep 02 2005