Western blot abreview for Anti-Myc tag antibody

Average
Abreviews
Application
Western blot
Sample
Human Cell lysate - whole cell (Jurkat, K562, and HL-60 cells)
Loading amount
50000 cells
Specification
Jurkat, K562, and HL-60 cells
Gel Running Conditions
Reduced Denaturing (4-20% Bis-Tris)
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 20°C

Other product details

Dilution
1/500
Incubation time
16 hour(s) and 0 minute(s) · Temperature: 4°C · Diluent: PBS + 0.05% Tween-20 + 5% BSA

Secondary antibody

Name
Non-Abcam antibody was used: goat anti-rabbit HRP
Host species: Goat
Clonality: Polyclonal
Conjugation: Horse Radish Peroxidase
Dilution
1/5000

Detection

Detection method
ECL+
Exposure
10 minute(s) and 0 second(s)
Bands
Specific: 65 kDa Non-specific: 55 kDa
Positive control
Jurkat cells, which we know express KLF12.

Additional data

Additional Notes
We wanted to see if KLF12 was expressed in various cancer cell lines, with Jurkat cells as a positive control. This Western gave a band at around 65 kDa, which we believe is the right band since we have seen it disappear with siRNA against KLF12. However, a non-specific band shows up around 55-58 kDa, which we have marked with a star on the picture.

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Submitted Mar 21 2013

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