Western blot abreview for Anti-VDAC/Porin antibody

Average
Abreviews
Application
Western blot
Sample
Human Cell lysate - whole cell (cancer-associated fibroblasts of prostate carcinom)
Loading amount
20 µg
Specification
cancer-associated fibroblasts of prostate carcinom
Gel Running Conditions
Reduced Denaturing (Invitrogen's NuPAGE 4-12%)
Blocking step
Milk as blocking agent for 16 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 4°C

Other product details

Dilution
1/400
Incubation time
16 hour(s) and 0 minute(s) · Temperature: 4°C · Diluent: PBS + 0.075% Tween20 + 5% milk powder

Secondary antibody

Name
Non-Abcam antibody was used: Goat-anti-rabbit IgG IRDye 800CW (Licor Bioscience
Host species: Goat
Clonality: Polyclonal
Conjugation: IRDye® 800CW
Dilution
1/10000

Detection

Detection method
fluorescence via Licor Odyssey
Exposure
4 minute(s) and 0 second(s)
Bands
Specific: 32 kDa Non-specific: ~70, ~64 kDa
Positive control
own sample and simultaneous detection of Vimentin in other colour channel

Additional data

Additional Notes
I tested the VDAC AB with two different lysate loading amounts: 20 ug in lanes 1, 3, & 5 and 30 ug lysate in lanes 2, 4 & 6. The AB was tested at three different concentrations and in two different blocking buffers.
In my setting it worked best in milk powder at the concentration of1/400. Unfortunately several bands came up and therefore suggesting that multiplexing is very difficult.
In summary, it's a solid, average quality antibody doing it's job but not as good as some other abcam antibodies.

Dr. Alexander Henke

Verified customer

Submitted Aug 03 2011

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