ANTI-HA-BIOTIN HIGH AFFINITY (3F10)

Quality

Function test: The Anti-HA-Biotin; High Affinity is function tested by Western blot analysis of a HA-tagged fusion protein.

Preparation Note

Sample Materials
Sample preparation: Prepare protein extracts containing the HA-tagged protein of interest using any of a variety of standard methods. The following lysis buffers have performed well and should be taken as guidelines:

• Bacterial extracts: 20 mM Tris, pH 8.0, 100 mM NaCl, cOmplete Protease Inhibitor Cocktail Tablets, followed by freeze-thaw.
• Mammalian extracts: 50 mM Tris, pH 7.5, 150 mM NaCl, 0.1% Nonidet P40, complete Protease Inhibitor Cocktail Tablets.
• Other cell lysis buffers may be more appropriate for individual applications. In general, to obtain optimal performance of the affinity matrix:
• Use protease inhibitors to reduce proteolytic activity. Use complete Protease Inhibitor Cocktail Tablets for most applications.
• Limit detergent to the lowest concentration levels necessary to obtain adequate cell lysis.

Working concentration: Working concentration of conjugate depends on application and substrate

The following concentrations should be taken as a guideline:

• Dot blot: 100 ng/ml
• ELISA: 100 ng/ml
• Western blot: 100 ng/ml

Reconstitution

Add 1 ml double-distilled water to a final concentration of 50 μg/ml.
Rehydrate for 10 minutes prior to use.

Other Notes

For life science research only. Not for use in diagnostic procedures.