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 an HRP-conjugated 6x-His Epitope Tag monoclonal antibody (Product # MA1-21315-HRP) at a dilution of 1:1000 overnight at 4°C on a rocking platform and washed in TBS-0.1% Tween-20. Chemiluminescent detection was performed using SuperSignal West Pico (Product # 34080).
|Tested species reactivity||Tag|
|Published species reactivity||Yeast|
|Host / Isotype||Mouse / IgG2b|
|Immunogen||6x His synthetic peptide.|
|Storage buffer||PBS, pH 7.2, with proprietary stabilizer|
|Storage Conditions||4° C|
|Tested Applications||Dilution *|
|Western Blot (WB)||1:500 - 1:2000|
* 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.
|Western Blot (WB)||See 1 publications below|
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.
Yeast hnRNP-related proteins contribute to the maintenance of telomeres.
MA1-21315-HRP was used in western blot to study the role of hnRNP-related proteins in telomere maintenance
|Lee-Soety JY,Jones J,MacGibeny MA,Remaly EC,Daniels L,Ito A,Jean J,Radecki H,Spencer S||Biochemical and biophysical research communications (426:12)||2012|