Application
Western blot
Sample
Human Cell lysate - whole cell (Human embryonic stem and HeLa cell lines)
Gel Running Conditions
Reduced Denaturing (10% SDS-PAGE gel)
Loading amount
15 µg
Specification
Human embryonic stem and HeLa cell lines
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 2% · Temperature: 25°C
Other product details
Incubation time
16 hour(s) and 0 minute(s) · Temperature: 4°C · Diluent: Tris-buffered saline + 0.1%Tween20
Dilution
1/1000
Secondary antibody
Dilution
1/4000
Name
Non-Abcam antibody was used: Goat anti-Rabbit IgG
Host species: Goat
Clonality: Polyclonal
Conjugation: Horse Radish Peroxidase
Host species: Goat
Clonality: Polyclonal
Conjugation: Horse Radish Peroxidase
Detection
Detection method
ECL
Exposure
30 second(s)
Bands
Specific: 54 kDa
Positive control
H7.s14 human embryonic stem cell (hESC) lysate was used in comparison to HeLa cell lysate.
Additional data
Additional Notes
I included a loading control using the same incubation conditions. The loading control primary antibody was Þ-Tubulin (9F3) Rabbit mAb from a different company.
Laemlli buffer was used for cell lysis and a PVDF membrane for western blotting.
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.
Joanne Lacey
Verified customer
Submitted May 22 2017