Western blot abreview for Anti-DDDDK tag antibody [M2] (HRP)

Excellent
Abreviews
Application
Western blot
Loading amount
10 µg
Gel Running Conditions
Reduced Denaturing (10)
Sample
Saccharomyces cerevisiae Cell lysate - whole cell (S. cerevisiae cell lysate)
Specification
S. cerevisiae cell lysate
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5%

Other product details

Incubation time
2 hour(s) and 0 minute(s) · Temperature: 22°C
Dilution
1/3000

Secondary antibody

Secondary antibody
None used

Detection

Detection method
ECL
Bands
Specific: 78 kDa
Exposure
45 second(s)
Positive control
Lane 1, 2, 3, 4, 5, 6

Additional data

Additional Notes
Antibody works well with no significant non-specific background. In the experiment the protein of interest is tagged with a single FLAG tag and expressed with a constitutive promoter. Sample 1, 2, 3, 4, 5, 6 are the lysates from the strains expressing the tagged protein. The experiment was done with cells grown to 1.5 OD units and lysed using glass beads with the lysis buffer containing MOPS, SDS, Urea and EDTA. 0.4 OD equivalent of cells were analyzed on the SDS-PAGE and western blotting was preformed with the Abcam anti-Myc antibody which is tagged with HRP (ab49763). Nitrocellulose membrane was used for the blotting. Highly recommend this.

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Submitted Mar 04 2014

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