Name: Biotin-X NTA [Biotin-X nitrilotriacetic acid, potassium salt] *CAS 856661-92-2*
Brand: AAT Bioquest
SKU: 3009
Price: 115 USD
Availability: InStock

Biotin-X NTA

Biotin-X nitrilotriacetic acid, potassium salt; CAS 856661-92-2

Biotin-X nitrilotriacetic acid (biotin-X NTA) is widely used to detect histidine-tagged proteins immobilized on nitrocellulose membranes. The NTA moiety of biotin-X NTA chelates Ni ion that is also chelated with histidine tags. The NTA-polyHis-complex can be detected using standard enzyme-linked streptavidin methods. Biotin-X NTA can be used to detect less than 0.1 pmol of histidine-tagged protein using a streptavidin'horseradish peroxidase conjugate and chemiluminescence techniques. Biotin-X NTA can be removed from the histidine-tagged protein at pH 4.8, allowing the blot to be reanalyzed with another probe. In combination with fluorescent avidin conjugates, this NTA biotin derivative can be used for detecting polyhistidine-containing biomolecules such as fusion proteins. This NTA Biotin derivative is a bifunctional reagent that is used to detect histidine-tagged proteins immobilized. The nitrilotriacetic acid is used to chelate a Ni(II) ion at four of its six coordination sites. The remaining two sites are available for binding to a histidine tag. The biotin functional group can then be detected using a streptavidin-horseradish peroxidase conjugate and chemiluminescence. Using this biotinylated nitrilotriacetic acid, it is possible to detect less than 0.11 pmol of histidine-tagged Escherichia coli RNA polymerase sigma70 subunit. This reagent is also able to specifically detect His-tagged sigma70 from a whole cell lysate following SDS-PAGE and transfer to nitrocellulose. The reagent can be dissociated from the His-tagged protein at pH 4.8 and the blot can be reprobed with a monoclonal antibody for detection of different proteins on the same blot.

Protocol

SDS

COA

Catalog	Size	Price	Quantity
3009	1 mg	Price

Calculators

Common stock solution preparation

Table 1. Volume of DMSO needed to reconstitute specific mass of Biotin-X NTA [Biotin-X nitrilotriacetic acid, potassium salt] *CAS 856661-92-2* to given concentration. Note that volume is only for preparing stock solution. Refer to sample experimental protocol for appropriate experimental/physiological buffers.

	0.1 mg	0.5 mg	1 mg	5 mg	10 mg
1 mM	139.669 µL	698.344 µL	1.397 mL	6.983 mL	13.967 mL
5 mM	27.934 µL	139.669 µL	279.337 µL	1.397 mL	2.793 mL
10 mM	13.967 µL	69.834 µL	139.669 µL	698.344 µL	1.397 mL

Molarity calculator

Enter any two values (mass, volume, concentration) to calculate the third.

Mass (Calculate)		Molecular weight		Volume (Calculate)		Concentration (Calculate)		Moles
mg	÷	715.98 g/mol	=	mL	x	mM	=	mmol

Citations

View all 4 citations: Citation Explorer

Malaria Detection with Cyclic Catch-and-Release Signal Amplification

Authors:

Jacobs, Michael Zachary

A low-cost, high-performance system for fluorescence lateral flow assays

Authors:

Lee, Linda G and Nordman, Eric S and Johnson, Martin D and Oldham, Mark F

Journal:

Biosensors (2013): 360--373

Implementation of P22 viral capsids as nanoplatforms

Authors:

Kang, Sebyung and Uchida, Masaki and O'Neil, Alison and Li, Rui and Prevelige, Peter E and Douglas, Trevor

Journal:

Biomacromolecules (2010): 2804--2809

Calcium-dependent activation of transglutaminase 2 by nanosecond pulsed electric fields

Authors:

Morotomi-Yano, Keiko and Yano, Ken-ichi

Journal:

FEBS Open Bio

References

View all 78 references: Citation Explorer

Evaluation of a new biotin-DOTA conjugate for pretargeted antibody-guided radioimmunotherapy (PAGRIT((R)))

Authors:

Urbano N, Papi S, Ginanneschi M, De Santis R, Pace S, Lindstedt R, Ferrari L, Choi S, Paganelli G, Chinol M.

Journal:

Eur J Nucl Med Mol Imaging. (2006)

A biotin-protein bond with stability in plasma

Authors:

Bogusiewicz A, Mock NI, Mock DM.

Journal:

Anal Biochem (2005): 98

Probing the interaction of the biotin-avidin complex with the relaxivity of biotinylated Gd-DTPA

Authors:

Langereis S, Kooistra HA, van Genderen MH, Meijer EW.

Journal:

Org Biomol Chem (2004): 1271

Immunoassay of total prostate-specific antigen using europium(III) nanoparticle labels and streptavidin-biotin technology

Authors:

Huhtinen P, Soukka T, Lovgren T, Harma H.

Journal:

J Immunol Methods (2004): 111

Biotin is endogenously expressed in select regions of the rat central nervous system

Authors:

McKay BE, Molineux ML, Turner RW.

Journal:

J Comp Neurol (2004): 86

Physical properties

Molecular weight	715.98
Solvent	DMSO

Storage, safety and handling

H-phrase	H303, H313, H333
Hazard symbol	XN
Intended use	Research Use Only (RUO)
R-phrase	R20, R21, R22
Storage	Freeze (< -15 °C); Desiccated
UNSPSC	12352200
CAS	856661-92-2

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