New: Direct Detection of 6His-tagged Protein directly on Membranes

Time saving, sensitive detection without any additional detection reagent (ECL) required

Anti-6His mouse monoclonal antibody clone 13/45/31-2 detects N- and C-terminally His-tagged proteines as well as internal His-tags with a high affinity.

The new HisDirect 60nm Gold conjugates detect His-tagged Protein in Western Blotting and Dot-Blot on nitrocellulose and PVDF membranes and can also be used in microbial colony and plate tests.

The unique HisDirect protocol with an incubation time of only 10 – 60 minutes is a one step protocol were no additional washing steps, detection reagent such as ECL or substrate incubation is required, making it a time and cost saving alternative to conventional blotting detection methods.

Fig.1: Comparison of conventional detection methods and HisDirect ready-to-use ready-to-use reagent using lysates of two High Five® insect cell culture expression approaches. The arrow denotes the target protein. Left: Coomassie staining (3 h); Middle: HRP-anti-Penta-His antibody with ECL reagent (4 h); Right: HisDirect ready-to-use Pmol-Range (20 minutes)

HisDirect is available as ready-to-use reagent and also as useful pads

Ordering #QuantityDescription
HisDirect-PA-010 10 Pads HisDirect His-Tag WB Staining Pads (10x) pmol/fmol-Range (RUO)
HisDirect-PA-030 30 Pads HisDirect His-Tag WB Staining Pads (30x) pmol/fmol-Range (RUO)
HisDirect-LP-010 10x10ml HisDirect His-Tag WB Staining Solution pmol-Range ready-to-use (RUO)
HisDirect-LF-010 10x10ml HisDirect His-Tag WB Staining Solution fmol-Range ready-to-use (RUO)

Quantification of DotBlots with HisDirect:

Quantification of poly-histidine labelled protein using HisDirect Ready-to-use reagent. Left: Dot blot for the quantitative determination of His6-labelled 11 kDa proteins. The red frame marks the linear detection range. Left: On a nitrocellulose strip, 0,5 μL each of a protein dilution series were incubated for 90 min in 10 mL HisDirect Ready-to-use reagent, dried and photographed, the photo converted into greyscale, and the amount of protein applied noted; Middle: Quantification with the aid of a dilution series. Correlation between signal intensity and the amount of protein applied determined from the greyscale image using the quantification program Image Studio Lite 5.2. Right: The linear range lies between a protein amount of 6,35 pmol and 50 pmol, or 69 ng and 0,55 μg, for areas of approx. one squaremillimeter.

Additional anti 6His Antibody Formats and Resources: