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HTRF 6His Detection Kit, 500 Assay Points
HTRF 6His Detection Kit, 500 Assay Points
Generic HTRF Anti tag XL665 image
| Feature | Specification |
|---|---|
| Application | Protein-Protein Interaction |
| Sample Volume | 10 µL |
Product variants
Unit Size: 500 assay points
Part #:
64HISPEG
Unit Size: 10,000 assay points
Part #:
64HISPEH
For research use only. Not for use in diagnostic procedures. All products to be used in accordance with applicable laws and regulations including without limitation, consumption, and disposal requirements under European REACH regulations (EC 1907/2006).
HTRF 6His Detection Kit, 500 Assay Points
Generic HTRF Anti tag XL665 image
HTRF 6His Detection Kit, 500 Assay Points
Product information
Overview
The kit enables the rapid detection of 6His tagged fusion proteins at each step of protein production and purification. It can either be used to ensure that the 6His motif of a fusion protein is accessible to the anti-6His antibody, or to approximate the concentration of a 6His-tagged protein.
Calibrated using the appropriate protein, the assay is very accurate and compatible with the SOPs, it can also be used to eliminate false positives and to predict the possibility of purifying the protein using a Nickel column: a non-detected Tag using our check kit will not be available for purification.
How it works
Assay principle
As shown in the graph, 6His peptide labeled with XL665 is detected by anti 6His Cryptate conjugate (anti-tag Cryptate). The 6His-tagged fusion protein (Tag fusion protein) to be ascertained competes with the 6His-peptide-XL665 conjugate (Tag-XL665) for binding to the anti-6His-Cryptate conjugate. The specific signal is inversely proportional to the concentration of 6His-tagged fusion protein.
Assay protocol
The example on the right describes the protocol using a 20 µL final assay volume. Dispense 10 µL of standard or of 6His-tagged protein of interest , with 5 µL of 6His Eu-Cryptate conjugate and 5 µL of 6His-XL665 conjugate into a well. Incubate for 2 hours at RT and read.
Assay validation
6His Check Kit Gold performance on various 6His-tagged proteins
The 6His check kit Gold enables the detection of 6His tagged proteins, whatever the molecular weight and the 6His tag insertion (N-terminal, C-terminal and internal 6His tags). The detection of the 6His tag varies for different fusion proteins.
For quantitative determination of 6His tagged protein in a sample, we recommended using an internal calibrator (6His fusion protein close to the 6His tagged protein of interest).
The 6His-MBP-GST-Flag and the THX-6His-1509V3 proteins were kindly provided by Hybrigenics.
| Tag insertion | 6His Tagged protein | MW(Da) | Quantification limit or IC10(nM) | IC50 (nM) |
|---|---|---|---|---|
| N-term | 6His-peptide | 1,300 | 4.2 | 32.8 |
| 6His BRD4(1) | 17,800 | 17.6 | 105.0 | |
| 6His-SNAP | 20,600 | 31.4 | 244.8 | |
| 6His-MBP-GST-Flag* |
64,000 | 3.2 | 28.0 | |
| Internal | THX-6His-1509V3* |
27,000 | 3.0 | 24.8 |
| C-term | 6His tagged Nanobody | 15,000 | 2.9 | 24.48 |
Specifications
| Application |
Protein-Protein Interaction
|
|---|---|
| Brand |
HTRF
|
| Detection Modality |
HTRF
|
| Product Group |
Kit
|
| Sample Volume |
10 µL
|
| Shipping Conditions |
Shipped in Dry Ice
|
| Target Class |
Binding Assay
|
| Technology |
TR-FRET
|
| Therapeutic Area |
Cardiovascular
Infectious Diseases
Inflammation
Metabolism/Diabetes
NASH/Fibrosis
Neuroscience
Oncology & Inflammation
Rare Diseases
|
| Unit Size |
500 assay points
|
Video gallery
HTRF 6His Detection Kit, 500 Assay Points
HTRF 6His Detection Kit, 500 Assay Points
Resources
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Brochure
HTRF assays and reagents product list
Discover the versatility and precision of Homogeneous Time-Resolved Fluorescence (HTRF) technology. Our HTRF portfolio offers a...
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