Description
The engineering of a short epitope tag to a recombinant protein provides a very convenient method for detecting and isolating the protein of interest. For example, the placement of a series of 6 consecutive histidine residues that are incorporated at either end of the recombinant fusion protein facilitates the purification of that protein. Purification can be easily accomplished by metal chelate affinity chromatography using a metal resin column. The 6xHis monoclonal antibody can be used to detect the epitope-tagged protein of interest.
An E. coli produced recombinant 6xHis control lysate is provided as a positive control. Store lysates at -20°C. Note: The size of the band observed will vary according to the size of the recombinant protein. In the example shown, the lysate contained a recombinant protein of ~31 kDa and the 6xHis tag does not appreciably change the observed molecular weight.
Store undiluted at 4°C. The monoclonal antibody was purified from tissue culture supernatant or ascites by affinity chromatography.
1. Since applications vary, each investigator should titrate the reagent to obtain optimal results.
2. Caution: Sodium azide yields highly toxic hydrazoic acid under acidic conditions. Dilute azide compounds in running water before discarding to avoid accumulation of potentially explosive deposits in plumbing.
3. Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
4. Sodium azide is a reversible inhibitor of oxidative metabolism; therefore, antibody preparations containing this preservative agent must not be used in cell cultures nor injected into animals. Sodium azide may be removed by washing stained cells or plate-bound antibody or dialyzing soluble antibody in sodium azide-free buffer. Since endotoxin may also affect the results of functional studies, we recommend the NA/LE (No Azide/Low Endotoxin) antibody format, if available, for in vitro and in vivo use.
5. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
