Application
Western blot
Sample
Human Cell lysate - whole cell (HEK293T cells)
Gel Running Conditions
Reduced Denaturing (5% stacking and 10% separating Bis-Tris gel)
Loading amount
270000 cells
Specification
HEK293T cells
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 22°C
Other product details
Incubation time
16 hour(s) and 0 minute(s) · Temperature: 4°C · Diluent: 5% milk 0.1% Tween in TBS buffer
Dilution
1/10000
Secondary antibody
Dilution
1/10000
Name
Non-Abcam antibody was used: IRDye® 800CW Goat anti-Rabbit IgG (H + L), 0.5 mg
Host species: Goat
Clonality: Polyclonal
Conjugation: IRDye® 800CW
Host species: Goat
Clonality: Polyclonal
Conjugation: IRDye® 800CW
Detection
Detection method
Near Infrared Fluorescence Imaging
Negative control
HEK293T PEX19 knockout cells
Exposure
1 second(s)
Bands
Specific: 33 kDa Non-specific: 10, 40, 45, 110 kDa
Positive control
WT HEK293T cells
Additional data
Additional Notes
We use Odyssey Near Infrared Fluorescence Imaging System. In this system the signal intensity is detected by scanning, and the scan intensity setting was 5. That is why, we do not have an exposure time.
Line 1) contains whole cell lysate of HEK293T; Line 2 and 3) contain whole cell lysate of HEK293T PEX19 CRISPR knockout 1 and 2, respectively. The predicted band size of PEX19 is 33 kDa.
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. Tamara Somborac
Verified customer
投稿 Feb 16 2018