Application
Western blot
Sample
Human Cell lysate - whole cell (Human Foreskin Fibroblasts & Mouse embryonic fibro)
Gel Running Conditions
Reduced Denaturing (4-12% Bolt Bis-Tris Plus gradient gel)
Loading amount
200000 cells
Specification
Human Foreskin Fibroblasts & Mouse embryonic fibro
Blocking step
Li-Cor Odyssey Blocking Buffer (TBS) as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 100% · Temperature: 25°C
Other product details
Incubation time
16 hour(s) and 0 minute(s) · Temperature: 4°C · Diluent: Li-Cor Odyssey Blocking Buffer (TBS)
Dilution
1/1000
Secondary antibody
Dilution
1/5000
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
Exposure
5 minute(s) and 0 second(s)
Positive control
Lane 1: HUMAN foreskin fibroblast cells
Lane 2: MOUSE embryonic fibroblasts (positive control of FADD detection)
Additional data
Additional Notes
We used both anti-FADD and anti-beta actin (ab6276) antibodies for the western blot. The 42 kDa green band is the beta actin band. The FADD antibody itself detected a 26 kDa red band in the mouse cell lysates but not the human one. This data clearly shows that the antibody only detect mouse FADD protein.
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.
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投稿 Feb 05 2016