Western blot analysis of 6x-His Epitope Tag was performed by loading various amounts of E. coli lysate containing a multi-epitope tagged protein per well onto a 4-20% Tris-HCl polyacrylamide gel. Proteins were transferred to a PVDF membrane and blocked with 5% BSA/TBST for at least 1 hour. The membrane was probed with a biotinylated 6x-His Epitope Tag monoclonal antibody (Product # MA1-21315-BTIN) at a dilution of 1:1000 for 1 hour at room temperature, followed by Streptavidin-HRP (Product # 21126) at a dilution of 1:20,000 for 30 minutes at room temperature. Chemiluminescent detection was performed using SuperSignal West Pico (Product # 34078).
|Tested species reactivity||Tag|
|Host / Isotype||Mouse / IgG2b|
|Immunogen||6x His synthetic peptide.|
|Storage buffer||PBS with proprietary stabilizer|
|Contains||0.02% sodium azide|
|Storage Conditions||4° C, do not freeze|
|Tested Applications||Dilution *|
|ELISA (ELISA)||Assay Dependent|
|Immunoprecipitation (IP)||Assay Dependent|
|Western Blot (WB)||1:500 - 1000|
* Suggested working dilutions are given as a guide only. It is recommended that the user titrate the product for use in their own experiment using appropriate negative and positive controls.
MA1-21315-BTIN has been successfully used for Western blot, ELISA, IF and IP
Epitope tags provide a method to localize gene products in a variety of cell types, study the topology of proteins and protein complexes, identify associated proteins, and characterize newly identified, low abundance or poorly immunogenic proteins when protein specific antibodies are not available. Expression of recombinant proteins in E. coli as a fusion protein with neighboring histidine residues is one of the most popular methods of epitope tags. The affinity of the histidine-tag motif to Ni2+ by chelation is strong and selective enough to enable purification of the protein to homogeneity by affinity chromatography on a Ni2+-NTA adsorbent.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.