Application
Western blot
Sample
Human Cell lysate - whole cell (Human primary airway epithelial and fibroblast cel)
Gel Running Conditions
Reduced Denaturing (12% gel)
Loading amount
30 µg
Specification
Human primary airway epithelial and fibroblast cel
Blocking step
Milk as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: RT°C
Other product details
Incubation time
24 hour(s) and 0 minute(s) · Temperature: 4°C · Diluent: 5% milk in TBST (0.05%)
Dilution
1/1000
Secondary antibody
Dilution
1/5000
Name
Non-Abcam antibody was used: Thermo FIsher 31460
Host species: Goat
Clonality: Polyclonal
Conjugation: Horse Radish Peroxidase
Host species: Goat
Clonality: Polyclonal
Conjugation: Horse Radish Peroxidase
Detection
Detection method
ECL+
Negative control
Cultured primary human airway fibroblast cells.
Exposure
5 minute(s) and 0 second(s)
Bands
Specific: ~70-75 kDa
Positive control
Cultured primary human airway epithelial cells.
Additional data
Additional Notes
Cells scraped in ice cold PBS, pelleted and resuspended in Roche cOmplete, EDTA-free protease inhibitor solution.
Samples were lysed by sonication, protein quantified by Pierce BCA protein assay.
Samples denatured at 100°C in presence of beta-mercaptoethanol for 5 minutes before loading.
Wet transferred to PVDF membrane for probing.
Lane 1 - Cultured primary human airway epithelial cells.
Lane 2 - Cultured primary human airway fibroblast cells.
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. Aaron Gardner
Verified customer
投稿 Oct 24 2016