Application
Western blot
Loading amount
30 µg
Gel Running Conditions
Reduced Denaturing (12%)
Sample
Human Cell lysate - whole cell (Caco-2, human colon epithelial cells)
Specification
Caco-2, human colon epithelial cells
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 22°C
Other product details
Concentration
1 µg/ml
Incubation time
16 hour(s) and 0 minute(s) · Temperature: 4°C · Diluent: 5% BSA in TBST
Secondary antibody
Name
Non-Abcam antibody was used: goat anti-mouse
Host species: Goat
Clonality: Polyclonal
Conjugation: Horse Radish Peroxidase
Host species: Goat
Clonality: Polyclonal
Conjugation: Horse Radish Peroxidase
Dilution
1/5000
Detection
Detection method
ECL
Bands
Specific: None kDa Non-specific: 50, 60 kDa
Exposure
10 minute(s) and 0 second(s)
Additional data
Additional Notes
•Buffer used for cell lysates ß Lysis buffer (TX-100): 20 mM Tris-Hcl, 150 mM NaCl, 10 mM NaF, 1 mM EDTA, 1% TX-100
•Protease/phosphatase inhibitors used ß Aprotonin, Leupeptin and Pepstatin (10 ´g/ml) ; 0.1 mM Sodium orthovanadate; 0.75 mM Pepstatin
•Samples boiled at 95C for 5 minutes
Abcam response
Thank you for your valuable feedback. We have been in contact with the customer and offered them a free of charge replacement or refund as stated by our Abpromise guarantee.
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.
MRS. Monika Schumacher
Verified customer
Submitted Feb 25 2015