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HTRF 6His Check kit Gold, 10000 Assay Points

The 6His check kit Gold enables the rapid detection of 6His tagged fusion proteins.

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 & disposal requirements under European REACH regulations (EC 1907/2006).

Product Variants
Part number: 64HISPEG
Unit Size: 500 Assay Points
List price: USD 1,039.00
Your price:
USD 0.00
USD 1,039.00 /each
Part number: 64HISPEH
Unit Size: 10,000 Assay Points
List price: USD 6,819.00
Your price:
USD 6,819.00
USD 6,819.00 /each

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.

Specifications

Assay Points
10000
Assay Target Type
Kit
Assay Technology
HTRF
Brand
HTRF
Quantity
1
Therapeutic Area
Cardiovascular
Infectious Diseases
Inflammation
Metabolism/Diabetes
NASH/Fibrosis
Neuroscience
Oncology & Inflammation
Rare Diseases
Unit Size
10,000 Assay Points

Video gallery

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.

6his-check-kit-gold

 

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.

6his-check-kit-gold

 

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.

6his-check-kit-gold

 

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

 

 

Resources

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HTRF solutions, guide to major applications

This guide provides you an overview of HTRF applications in several therapeutic areas.

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