Product Information

Contents: eFluor^TM 490^NC (Carboxyl)
Catalog Number: 90-6364
Sizes: 200 ul
Formulation: 10 μM aqueous solution
Storage Conditions: Store at 4°C.
DO NOT FREEZE.
LIGHT-SENSITIVE MATERIAL.

Available Formats of This Product
Cat. No.	Format	Excite (nm)	Emit (nm)	Reported Applications
90-6364	eFluor^TM 490^NC (Carboxyl)	350-500	490	Biomolecule Conjugation
91-6364	eFluor^TM 525^NC (Carboxyl)	350-500	525	Biomolecule Conjugation
93-6364	eFluor^TM 605^NC (Carboxyl)	350-500	605	Biomolecule Conjugation
94-6364	eFluor^TM 625^NC (Carboxyl)	350-500	625	Biomolecule Conjugation
95-6364	eFluor^TM 650^NC (Carboxyl)	350-500	650	Biomolecule Conjugation
96-6364	eFluor^TM 700^NC (Carboxyl)	350-500	700	Biomolecule Conjugation Cell Labeling In vivo Imaging

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Description

Reactive eFluor™ Nanocrystals (Carboxyl), composed of either CdSe or cadmium-free InGaP, are functionalized nanocrystals that can be easily conjugated to proteins, oligonucleotides and other biomolecules of interest using simple binding chemistries and easy-to-complete protocols. The straightforward procedures are designed to allow the user to apply eFluor™ Nanocrystal technology in a wide variety of assay types, such as immunoassays, protein interaction assays, and other fluorescence-based detection assays.

Applications Reported

For research use only, not for diagnostic or therapeutic use. This product has been reported for use in conjugation to biomolecules.

Related Products

Cat. 91-6364  eFluor™ 525NC (Carboxyl)
Cat. 93-6364  eFluor™ 605NC (Carboxyl)
Cat. 94-6364  eFluor™ 625NC (Carboxyl)
Cat. 96-6364  eFluor™ 700NC (Carboxyl)
Cat. 90-6366  eFluor™ 490NC (Amine)
Cat. 91-6366  eFluor™ 525NC (Amine)
Cat. 93-6366  eFluor™ 605NC (Amine)
Cat. 94-6366  eFluor™ 625NC (Amine)
Cat. 96-6366  eFluor™ 700NC (Amine)

Reactive eFluor™ Nanocrystal (Carboxyl) Conjugation Protocol

Materials

2nmol eFluor™ Nanocrystals (Carboxyl) (8-10 uM)
20X Borate Buffer (Pierce, cat. 28341): prepare 1X Borate Buffer in dH2O (50 mM Borate, pH8.5)
50 mM Borate Buffer, pH7.2
EDC (1-ethyl-3-[3-dimethylaminopropyl]carbodiimide hydrochloride) (Pierce, cat. 22980)
- Prepare 50 mM EDC solution in dH2O immediately before adding to eFluor™ Nanocrystals (Carboxyl).
Biomolecule of interest
Note: Biomolecule of interest must be free of preservatives, such as BSA and azide. Biomolecule solution must be amine free (50 mM Borate buffer, pH7.2 is recommended).
Glass vials
Ultrafiltration units with 100 kDa molecular weight cutoff (Millipore, Amicon Ultra-15, cat. UFC910096)
Note: If biomolecule is <60 kDa use ultrafiltration to separate un-reacted biomolecules from eFluor™ Nanocrystals (Carboxyl) conjugates
Superdex 200 (GE Healthcare, cat. 17-1043-10 or cat. 17-5175-01)
Note: If biomolecule is >60 kDa use HPLC or FPLC for optimal separation of un-reacted biomolecules from eFluor™ NC conjugates

Method

eFluor™ Nanocrystal (Carboxyl) Activation
1. Transfer 2.0 nmoles of eFluor™ Nanocrystals (Carboxyl) to a glass vial and activate by adding 50 uL of 50mM EDC dissolved in dH2O. Mix well. There is no need to incubate at this point.
2. Add appropriate amount of biomolecule to the reaction mixture. Optimal coupling ratio may vary depending on biomolecule and must be determined by the end user.
3. Incubate the reaction mixture for 2 hours at room temperature while gently mixing.
Conjugate Purification
1. Biomolecules < 60kDa:
  Add the conjugate to an Ultrafiltration unit with 100 kDa molecular weight cutoff and wash 5-6 times with 1X Borate buffer (pH8.5). Concentrate conjugate to <500µl.
  
  Biomolecules >60kDa:
  Purify the conjugate by FPLC or HPLC size exclusion chromatograpy using a Superdex 200 matrix equilibrated with 1X Borate buffer (pH8.5). Pool the fluorescent fraction (use a black light to identify fluorescent fractions), and concentrate to < 500ul using an Ultrafiltration unit with 100 kDa molecular weight cutoff. Pool the fractions containing free biomolecule and quantify the amount of un-reacted material to determine the coupling efficiency.