Western blot abreview for Anti-DDDDK tag antibody [F-tag-01] - (Equivalent to FLAG antibodies from Sigma)

Average
Abreviews
Application
Western blot
Sample
Human Purified protein (FLAG tagged purified SPOCK2 protein)
Gel Running Conditions
Reduced Denaturing
Loading amount
0.001 µg
Specification
FLAG tagged purified SPOCK2 protein
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 23°C

Other product details

Incubation time
2 hour(s) and 0 minute(s) · Temperature: 23°C · Diluent: same as blocking buffer
Dilution
1/1000

Secondary antibody

Dilution
1/3000
Name
Non-Abcam antibody was used: ECL anti mouse IgG HRP linked while antibody
Host species: Sheep
Clonality: Polyclonal
Conjugation: Horse Radish Peroxidase

Detection

Detection method
ECL
Negative control
With FLAG tagged purified proteins, negative control is not needed. Only 1 band on the blot per lane is possible.
Exposure
30 second(s)
Bands
Specific: 50 kDa
Positive control
The exact same protein and the exact same amount was loaded, but probed with rabbit anti-FLAG from Sigma on a distinct nitrocellulose filter. 1:1000 dilution was used for both primaries.

Additional data

Additional Notes
This antibody clearly works for a western blot, although affinity appears not as strong as the rabbit anti-FLAG from Sigma. To make the uploaded photograph, the two lanes probed with distinct antibodies was spliced to be adjacent in Photoshop. It is possible the difference in band intensity is due to the unique secondary antibodies that had to be used for each primary, since the primaries were made in different species.

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Submitted Nov 19 2018

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