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His Tag ELISA Detection Kit

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Overview

Description

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.

In optimized 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.

Key Features

Features	Specifications
Sensitivity	1 ng/ml His-tagged proteins
Detection Range	1 ng/ml~729 ng/ml
Test Samples	N-terminal/C-terminal/internal His-tagged proteins. 4 x His/5 x His/6 x His-tagged proteins. Mammalian, 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)

Product Information

Component	Quantity	Part. No
His Tag Plate	1 plate (8 wells x 12 strips)	436-80
Anti-His Monoclonal Antibody	6 ml	436-20
Antibody Tracer	12 ml	436-30
His Tag Standards (0,1,3,9,27,81,243,729 ng/ml)	1 ml per each standard	436-11,12,13,14,15,16,17,18
Assay Diluent	60 ml	436-60
20 × Wash Solution	40 ml	436-70
TMB Substrate	12 ml	436-40
Stop Solution	6 ml	436-50
His Tag Standard Stock (10 μg/ml)	500 μl	436-10
Plate Sealer	2	N/A
User Manual	1	N/A

Properties

Storage	The unopened kit is stable for at least 12 months if stored at 2-8 °C and the opened kit may be stable for up to 1 month at 2-8 °C. Do not freeze the kit.
Note	For Research Use Only. Not for Use in Diagnostic Procedures. The operator should read technical manual carefully before using this product.

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