Biotin-X NTA [Biotin-X nitrilotriacetic acid, potassium salt] CAS 856661-92-2

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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

Overview SDS Protocol

Example protocol

PREPARATION OF WORKING SOLUTION

1. Biotin-X NTA working solution (1mg/mL):
Add 1 mL of DMSO or ddH₂O to Biotin-X NTA vial and mix well.

2. Staining Solution:
Prepare fresh Staining Solution (less than 30 minutes before use): prepare 20 mL of Staining Solution for each 8 cm ×10 cm blot by adding 20 μL of 10 mM NiCl₂ , 20 μL of 1 mg/mL biotin-X NTA working solution and 1-2 μL of 1 mg/mL streptavidin–alkaline phosphatase to 20 mL of Blocking Buffer. Mix well. Note: Once the solution is made, it is ready to use right away, and remains stable for at least 30 minutes.

SAMPLE EXPERIMENTAL PROTOCOL

Sample Protocol for detection of His-tagged Protein in PVDF

Prepare your PVDF blots as needed (block nonspecific binding sites and wash the blot).
Incubate the blot with 20 mL of Staining Solution at room temperature for 30 minutes.
Wash the blots, and pursuit for chemiluminescence detection.

Sample Protocol for Biotin-X NTA–Probed Blot Stripping

If desired, the blot can be stained using other detection methods.
For staining with antibodies or lectins, strip the biotin-X NTA complex off the blot by incubating it in 62.5 mM Tris, 0.2% SDS, 50 mM dithiothreitol (DTT), pH 6.8, at 50°C for 40 minutes with gentle agitation. Or incubated overnight in 200 mM acetate acid with 40 mM EDTA, pH 4.8.
Wash the blot in Wash Buffer at room temperature 2-4 times for 5 minutes each and proceed with antibody detection.

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
	/		=		x		=

Images

Figure 1. Chemical structure for Biotin-X NTA [Biotin-X nitrilotriacetic acid, potassium salt] *CAS 856661-92-2*

Citations

View all 4 citations: Citation Explorer

Malaria Detection with Cyclic Catch-and-Release Signal Amplification
Authors: Jacobs, Michael Zachary
Journal: (2017)

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

Instability of the biotin-protein bond in human plasma
Authors: Bogusiewicz A, Mock NI, Mock DM.
Journal: Anal Biochem (2004): 156

Design, synthesis, and evaluation of 5'-S-aminoethyl-N(6)- azidobenzyl-5'-thioadenosine biotin conjugate: a bifunctional photoaffinity probe for the es nucleoside transporter
Authors: Addo JK, Buolamwini JK.
Journal: Bioconjug Chem (2004): 536

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

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

Volume of RBCs, 24- and 48-hour posttransfusion survivals, and the lifespan of (51)Cr and biotin-X-N-hydroxysuccinimide (NHS)-labeled autologous baboon RBCs: effect of the anticoagulant and blood pH on (51)Cr and biotin-X-NHS elution in vivo
Authors: Valeri CR, Pivacek LE, Cassidy GP, Ragno G.
Journal: Transfusion (2002): 343

Endogenous biotin staining in a subset of spinal neuronal cell bodies: a potential confounding factor for neuroanatomical studies
Authors: Berkowitz A., undefined
Journal: Brain Res (2002): 98

End-labeling of long DNA fragments with biotin and detection of DNA immobilized on magnetic beads
Authors: Xu H, Zhang S, Liu D, Liang CC.
Journal: Mol Biotechnol (2001): 183