Application
Western blot
Sample
Human Cell lysate - whole cell (ARPE-19)
Gel Running Conditions
Reduced Denaturing
Loading amount
15 µg
Treatment
125 ug/ml Cigarette Smoke Extract
Specification
ARPE-19
Blocking step
BSA 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
Dilution
1/400
Secondary antibody
Dilution
1/5000
Name
Non-Abcam antibody was used: Amersham ECL Rabbit IgG, HRP-linked whole Ab (from
Host species: Donkey
Clonality: Polyclonal
Conjugation: Horse Radish Peroxidase
Host species: Donkey
Clonality: Polyclonal
Conjugation: Horse Radish Peroxidase
Detection
Detection method
ECL+
Exposure
3 minute(s) and 0 second(s)
Bands
Specific: 51 kDa Non-specific: 45, 38 kDa
Positive control
cigarette smoke treatment
Additional data
Additional Notes
1 – DMSO 24h
2 – CSE 24h
3 – DMSO 12h
4 – CSE 12h
5 – DMSO 6h
6 – CSE 6h
7 – DMSO 3h
8 – CSE 3h
9 – DMSO 1h
10 – CSE 1h
11 – DMSO 0.5h
12 – CSE 0.5h
Multiple bands were observed, among which, the 51kDa band might be pGSK3alpha, we also observed 45 kDa and 38 kDa non specific bands.
Using the same samples we examined pGSK3beta using #ab75745, which gave a clear single 47 kDa band. Thus, we conclude the #ab4797 is good in detecting neither pGSK3alpha nor pGSK3beta.
Abcam response
Our Scientific Support team is currently investigating this negative Abreview, and we will resolve the issue in accordance with our Abpromise.
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. Lei Wang
Verified customer
投稿 Jul 25 2017