His-tag is one of the most widely applied tags for recombinant protein expression and
purification. It is also used for binding assay to detect protein-protein interactions by a pull-down assay
or in conjunction with anti-His antibody as IP/Co-IP assay. The three new high quality mAb based assay
products will provide convenient, easy, and robust tools for different purposes, including purification and
detection of His-tagged fusion-protein for downstream assays, enrichment and screening of His-tagged
fusion-protein mutants, and Co-IP & screening of interacting proteins.
Applications
Detect His-tagged proteins
Screen optimal expression conditions
Semi-quantitation with known His-tagged protein as standards
His Tag ELISA Detection Kit
GenScript His Tag Protein Detection Kit is developed for the detection of His-tagged protein
expression and
its relative quantification using His-tagged protein standards of known molecular weight (M.W.). This kit is
based on competitive ELISA, suitable for most systems, such as bacterial, yeast, mammalian cell lysates and
cell culture supernatant. This kit also offers a His-tagged protein stock for customer recovery verification
assay.
Figure 1. His-tagged protein standard
curve In accordance with the evaluation method outlined in the , the
Quantitation Limit for the His tag standard in this kit is 1 ng/mL, with a linear
range of 1 to 243 ng/mL.
Table 1. A typical sample data for His-tagged protein standard
curve
His Tag Standard (ng/mL)
Absorbance (OD 450 nm)
Measured His Tag Standard (ng/mL)
CV
Recovery Rate
Duplicate 1
Duplicate 2
Average
0
2.6672
2.7283
2.6978
/
/
/
1
2.3134
2.2391
2.2763
1.16
15%
116%
3
1.8494
1.9124
1.8809
2.69
8%
90%
9
1.1055
0.9926
1.0491
9.64
12%
107%
27
0.5196
0.5107
0.5152
26.88
2%
100%
81
0.2486
0.2621
0.2554
70.41
5%
87%
243
0.1241
0.1184
0.1213
272.77
8%
112%
729
0.0863
0.0823
0.0843
/
/
/
*Note: The concentration of 729 ng/mL
His-tag standard is the AP (anchor point) used to enhance the fit of the standard curve,
and it is not within the quantitative range of the kit.
Table 2. Intra-assay and inter-assay precision of the kit
Quality Control
His-tagged protein (ng/mL)
Intra-assay (n=10)
Inter-assay (n=30)
Average Measured His-tagged protein (ng/mL)
CV %
Recovery Rate
Average Measured His-tagged protein (ng/mL)
CV %
Recovery Rate
Precision 1
3
2.83
7.3%
94%
3.02
11.5%
101%
Precision 2
27
27.79
7.4%
103%
27.56
3.6%
102%
Precision 3
100
104.50
9.5%
105%
105.04
5.0%
105%
The intra-assay precision of the L01010 detection kit is less than 10%, and the
inter-assay precision is less than 15%.
His Tag Antibody Plate
His Tag Antibody Plate is a microtiter plate coated with THE™ Anti His Tag Antibody. The
plate is
designed for capturing His tagged protein with His tag located at N-terminus, C-terminus, or internal of the
protein. It can be applied for analysis of different length of His tag including Hexa-His (6 His), Penta-His
(5-His) and Tetra-His (4-His).The plate demonstrate high sensitivity and high binding capacity. The capacity
and sensitivity varies, depending on protein size, structure and solution buffer. Generally, proteins with
lower molecular weight (M.W) can be more sensitive to the His antibody plate than ones with higher M.W.
Compared with Nickel Coated Plate, the affinity plate can bind target proteins with high specificity, and
tolerable with several common used reagents such as EDTA, imidazole and β-ME.
L00436 is designed for a 96-well plate, suitable for direct transfer testing of 96-well
samples, but it is not suitable for the detection of rat and mouse samples.
L01010 is designed for a 384-well plate, suitable for large-scale sample testing or direct
transfer testing of 384-well samples, and it is suitable for the detection of mouse and rat
samples without matrix interference issues.
L01010 is a semi-quantitative kit. If precise quantitative analysis is required, it is
recommended to use your own standards and establish the standard curve according to the
guidelines provided with the kit.
For the detection and quantification of His-tagged proteins
For capturing and detecting His-tagged proteins
Steps
Involving multiple steps and the use of reagents
Simplify the steps for capturing His-tagged proteins
Product List
L00436_His Tag ELISA Detection Kit (L00436)
L01010_His Tag ELISA Detection Kit (384-Well Plate)
L00440B_His Tag Antibody Plate (black, 96 wells, nonremovable)
L00440C_His Tag Antibody Plate (clear, 8x12 strips)
L00440W_His Tag Antibody Plate (white, 96 wells, nonremovable)
Other Tagged Protein Detection & Purification
DYKDDDDK(Flag) Tag Antibody Plate (96-well)
DYKDDDDK (Flag) Tag Antibody Plate is a microtiter plate coated with THE™ Anti DYKDDDDK
(Flag) Tag
Antibody. The plate is designed for the rapid capturing of DYKDDDDK (Flag) tagged protein expressed in
different systems, including, E. coli, yeast and mammalian extracts and cell culture supernatants. It can be
applied for the detection of DYKDDDDK (Flag) tag located at N-terminus, C-terminus, or internal of the
protein. The plate demonstrates high sensitivity and high binding capacity. The capacity and sensitivity
varies, depending on protein size, structure and solution buffer. Generally, proteins with lower molecular
weight (M.W) can be more sensitive to the DYKDDDDK (Flag) antibody plate than ones with higher M.W. The
plate
is tolerable with several common used reagents. Avoid using detection reagents that can recognize mouse IgG,
such as anti mouse IgG antibodies and protein A.