||GenScript His Tag ELISA Detection Kit, a 1.5 hour competition ELISA, is developed for rapid and high throughput detection of His-tagged
proteins. There are several potential usages:
- Quickly identify the presence of His-tagged proteins in samples.
- Optimize protein expression by monitoring the His-tagged proteins level.
- High throughput screening of stable cell lines expressing His-tagged proteins.
This kit is based on competitive ELISA method. His Tag Plate in the kit is a 96-well microtiter
plate coated with a His-tagged protein with the molecular weight of 12.7 kDa. The
plate, which comprises of 8 wells x 12 strips, is demountable. When Anti-His Monoclonal
Antibody and His-tagged proteins are added to the well , the coated His-tagged
proteins compete with His Tag Standard (His-tagged protein with the molecular weight of 11.3
kDa) in solution or His-tagged
protein in sample to interact with Anti-His Monoclonal Antibody. The higher the
concentration of the His-tagged protein in solution is, the less the antibody
bound to the plate will be. The Anti-His Monoclonal Antibody used in the kit is
a mouse anti His tag monoclonal antibody (GenScript A00186). Antibody Tracer, which is a horseradish peroxidase (HRP) conjugated Goat anti mouse IgG, is used for enzyme reaction. His-tagged protein, Anti-His Monoclonal Antibody and antibody tracer form a complex. Other unbound molecules can be removed by washing
solution. The antibody tracer reacts with TMB substrate to develop blue
product that turns yellow immediately when the Stop Solution is added, which
can be measured by microplate reader at 450 nM.
test condition, each absorbance value is indicated to the individual His-tagged
proteins amount in solution. His-tagged
protein standards of known concentration and the corresponding absorbance
values are used to construct a standard curve. With the standard curve, His-tagged
protein amount present in the unknown sample can be calculated by transforming
its absorbance value.
ng/ml His-tagged proteins|
His/5 x His/6 x His-tagged proteins.
yeast and bacteria cell lysates or cell supernatant.
|Conveniency||Provide all reagents required for test.|
Complete test within 1.5 hours.
|Reagent Compatibility||Tolerable with various reagents at certain
concentration (see Table 3)|
plate (8 wells x 12 strips)||436-80|
per each standard||436-11,12,13,14,15,16,17,18|
|His Tag Standard Stock (10 μg/ml)||500 μl||436-10|