Application
Western blot
Sample
Mouse Tissue lysate - whole (Retinal)
Loading amount
10 µg
Specification
Retinal
Gel Running Conditions
Reduced Denaturing (10% Tris Glycine Gel)
Blocking step
1:1 PBS: Licor Blocking buffer as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 50% · Temperature: 22°C
Other product details
Dilution
1/1000
Incubation time
14 hour(s) and 0 minute(s) · Temperature: 4°C · Diluent: 1:1 PBS: Licor Blocking buffer
Secondary antibody
Name
Non-Abcam antibody was used: IRDye 800CW Goat anti-Rabbit IgG (H + L)
Host species: Rabbit
Clonality: Polyclonal
Conjugation: IRDye® 800CW
Host species: Rabbit
Clonality: Polyclonal
Conjugation: IRDye® 800CW
Dilution
1/10000
Detection
Detection method
Licor Odyssey Scanner
Exposure
3 minute(s) and 0 second(s)
Bands
Specific: 40 kDa Non-specific: 52, 37, 30 kDa
Positive control
WT Mouse Retinal Cells (lane 1)
Additional data
Additional Notes
Lane 1 is MW ladder. From the top: 120, 88, 50, 34, 26. Lane 2,3, and 4 Retinal Lysate from 1 month old Mouse eyes of WT, KO, and heterozygote for PhLP gene, which should cause a decrease in levels of Galpha. Bands seen at 52 kDa, 40 kDa (in boxes). Other minor bands at ~37, ~30 kDa. The quantification shows that the 52 kDa bands are roughly the same but the KO 40 kDa band is significantly reduced. I suggest that this is the actual GNAT2 mouse band, which also matches the approximate MW of 40 kDa. The 52 kDa band is a non-specific band.
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.
Dr. Chris Tracy
Verified customer
投稿 Aug 24 2012