Application
Western blot
Sample
Dog Cell lysate - whole cell (thyroid cancer cell line)
Gel Running Conditions
Reduced Denaturing (NuPage 4-12% Bis-Tris Gel)
Loading amount
12.6 µg
Treatment
0.05-2uM of lapatinib and neratinib, 1% DMSO for 6hrs
Specification
thyroid cancer cell line
Blocking step
SuperBlock T20 (TBS) Blocking Buffer - **4mL** as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 4% · Temperature: 20°C
Other product details
Incubation time
18 hour(s) and 0 minute(s) · Temperature: 4°C · Diluent: SuperBlock T20 ITBS) Blocking Buffer
Dilution
1/1000
Secondary antibody
Dilution
1/40000
Name
Non-Abcam antibody was used: Goat anti-Rabbit IgG (H+L) Secondary Antibody, HRP
Host species: Goat
Clonality: Polyclonal
Conjugation: HRP polymer
Host species: Goat
Clonality: Polyclonal
Conjugation: HRP polymer
Detection
Detection method
ECL+
Exposure
5 minute(s) and 0 second(s)
Bands
Specific: 180 kDa Non-specific: 120 kDa
Positive control
SkBr3 Human Breast Carcinoma cell line, treated with 1% DMSO 6hrs.
Additional data
Additional Notes
There is nonspecific binding (~120kDa) that does not go away with suppression of HER2, while the known observed band (180kDa) does.
WB specifics: lysis buffer cocktail (M-PER, SDS, cOmplete mini tablet, NaOVA, PMSF), Immobilon transfer membrane PVDF.
Abcam response
We would like to thank this researcher for submitting their review. This antibody had not been tested for cross reactivity with dog and we were unsure of what the results would be. The reviewer received an AbTrial discount code for using the antibody in an untested species.
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.
Abcam user community
Verified customer
投稿 Feb 05 2021