Application
Western blot
Sample
Mouse Cell lysate - whole cell (mouse embryonic fibroblasts)
Gel Running Conditions
Reduced Denaturing
Loading amount
10000 cells
Treatment
With or without 100uM zVAD
Specification
mouse embryonic fibroblasts
Blocking step
Li-Cor Odyssey Blocking Buffer (TBS) as blocking agent for 14 hour(s) and 0 minute(s) · Concentration: 100% · Temperature: 4°C
Other product details
Incubation time
1 hour(s) and 0 minute(s) · Temperature: 25°C · Diluent: Li-Cor Odyssey Blocking Buffer (TBS)
Dilution
1/1000
Secondary antibody
Dilution
1/20000
Name
Non-Abcam antibody was used: IRDye 680RD Goat anti-Rabbit IgG
Host species: Goat
Clonality: Polyclonal
Conjugation: IRDye® 680
Host species: Goat
Clonality: Polyclonal
Conjugation: IRDye® 680
Detection
Detection method
Near-Infrared Fluorescence Imaging (Li-C
Exposure
10 minute(s) and 0 second(s)
Bands
Specific: 26-28 kDa Non-specific: 42 (should be beta-a kDa
Positive control
1st lane: wild-type MEFs without 100uM zVAD for 5 hours
2nd lane: wild-type MEFs with 100uM zVAD for 5 hours
Additional data
Additional Notes
We used both anti-FADD and anti-beta actin (ab6276) antibodies for the western blot. The 42 kDa band is the protein band of beta actin un-specifically recognized by secondary antibody. The anti-FADD antibody itself is specific to mouse FADD protein (26-28 kDa 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. Yi-Chieh Perng
Verified customer
提交于 Nov 30 2015