His tagged protein detection & purification

His tagged protein detection & purification

Catalog Products » Antibodies » Protein Detection and Immunoassay » His Tagged Protein Detection & Purification

Overview

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.

L00436

His Tag ELISA Detection Kit

$328.00 1 Kit

L01010

His Tag ELISA Detection Kit (384-Well Plate)

$1000.00 384 Tests

Selection Guide

Assay Principal

Competitive ELISA

Working Range

1-243 ng/mL

1 -729 ng/mL

Sensitivity

1 ng/mL

Throughput

384 tests

96 tests

Detection time

Complete assay in less than 1 hours and 15 minutes.(Incubation for 55 minutes)

Complete assay in less than 1 hour and 35 minutes.(Incubation for 1 hours and 15 minutes)

R2

R2≥0.98

Application Data

Figure 1. His-tagged protein standard curveIn 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

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

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

Precision 1

2.83

7.3%

94%

3.02

11.5%

101%

Precision 2

27.79

7.4%

103%

27.56

3.6%

102%

Precision 3

100

104.50

9.5%

105%

105.04

5.0%

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.

L00440B

His Tag Antibody Plate (black, 96 wells, non-removable)

$179.00 5 plates

L00440C

His Tag Antibody Plate (clear, 8 wells × 12 strips)

L00440W

His Tag Antibody Plate (white, 96 wells, non-removable)

His Tag Antibody Plate Selection Guide

Difference between Plates

Selection Guide

The key differences between L00440B, L00440W, and L00440C （See the table below for details）

Optical Principle

High light transmission (>90%)

Reflects light to enhance weak signals

Absorbs stray light to minimize background

Optimal Use

Absorbance (OD) detection

Chemiluminescence, low-intensity fluorescence, or reflectance-based colorimetric assays

High-sensitivity fluorescence (e.g., TRF, Cy5), low-background applications

Avoid

Fluorescence/chemiluminescence

High-background fluorescence assays

Absorbance detection

Choose different ELISA plates according to different application scenarios (see the table below for selection guide)

Colorimetric assays(e.g., HRP-TMB OD450/620)

L00440C (Clear)

Maximizes light transmission for accurate absorbance readings.

Chemiluminescence(e.g., Luciferase reporter)

L00440W (White)

Reflects emitted light to amplify weak signals (e.g., low-abundance targets).

Low-intensity fluorescence(e.g., FITC/TRITC)

Reflective background enhances signal-to-noise ratio for weak fluorescence.

High-sensitivity fluorescence(e.g., Cy5, TRF, FRET)

L00440B (Black)

Absorbs ambient and cross-well light, reducing background interference.

Flexible small-scale experiments

8x12 removable strips enable cost-effective testing.

FAQ

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.

Coated Materials

His tag protein

Anti-His antibody

Component

Antibody, substrate, enzyme

Pre-coated anti-His antibody plate

Application

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)

Incompatible plates compromise data integrity:

1. Clear plates for fluorescence/chemiluminescence: Light leakage between wells → Increased crosstalk and background noise (SNR drops >50%).False positives/negatives due to signal contamination.

2. Black plates for absorbance assays: Light absorption by black walls → Underestimated OD values (e.g., TMB OD450 may drop by 70%).

3. White plates for absorbance assays: Suboptimal light transmission → Reduced sensitivity.

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.

L00455B

DYKDDDDK Tag Antibody Plate (black, 96 wells, non-removable)

$229.00 5 plates

L00455C

DYKDDDDK Tag Antibody Plate (clear, 8 wells × 12 strips)

L00455W

DYKDDDDK Tag Antibody Plate (white, 96 wells, non-removable)

DYKDDDDK Tag Antibody Plate Selection Guide

The key differences between L00455B, L00455W, and L00455C（See the table below for details）

L00455C (Clear)

L00455W (White)

L00455B (Black)

Related products

A00174

His-tag Antibody, pAb, Rabbit

$102.00 40 μg

A00186

THE™ His Tag Antibody, mAb, Mouse

$385.00 100 μg

A00205

His-tag antibody [Biotin], pAb, Rabbit

A00612

THE™ His Tag Antibody [HRP], mAb, Mouse

$425.00 100 μg

A00613

THE™ His Tag Antibody [Biotin], mAb, Mouse

A01620

THE™ His Tag Antibody [FITC], mAb, Mouse

L00223

High Affinity Ni-Charged Resin

$83.00 10 ml Resin (Total 20 ml) 25 ml Resin (Total 50 ml) 500 ml Resin (Total 1000 ml)

L00295

Ni-charged MagBeads

$101.00 8 ml (2 ml Settled Beads)

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