Western blot abreview for Anti-IKK alpha (phospho S176 + S180) antibody

Below Average
Abreviews
Application
Western blot
Sample
Pig Tissue lysate - whole (Intestine (colon, jejunum and Ileum))
Loading amount
50 µg
Specification
Intestine (colon, jejunum and Ileum)
Gel Running Conditions
Reduced Denaturing (10%)
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 22°C

Other product details

Dilution
1/1000
Incubation time
2 hour(s) and 0 minute(s) · Temperature: 22°C

Secondary antibody

Name
Non-Abcam antibody was used: Anti-rabbit IgG (Cell Signaling - cod: #7064)
Host species: Goat
Clonality: Polyclonal
Conjugation: Horse Radish Peroxidase
Dilution
1/5000

Detection

Detection method
ECL+
Exposure
30 second(s)
Bands
Specific: 85 kDa Non-specific: many kDa

Additional data

Additional Notes
I made a test running different amount of protein (10, 20, 30, 40 and 50 µg of protein) in the same gel.
I had a lot non-specific binding and I tried different dilutions with the secondary antibody and did not work better.

It was used for the WB: PVDF menbrane, the Complete Protease Inhibitor Cocktail from Roche (cat no: 04693/24001), Lysis buffer from thermo Scientfic (Tissue protein extractio, cat no: 78510), ECL from Pierce (cat n°: PI32106) and Film Hyblot from Denville Scientific (cat no: E3012).
Abcam response
This customer received a testing discount to try this antibody in pig, an untested species. The preliminary data suggest that this antibody may not be suitable for detection of the pig protein.

Ms. Ananda Rodrigues

Verified customer

Submitted Aug 02 2010

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