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NIR Fluorochrome Labeling Kit
Caution: For Laboratory Use. A product for research purposes only.

VivoTag™ 680XL Protein Labeling Kit
Product Number: NEV11118
DESCRIPTION
PerkinElmer’s VivoTag 680XL Protein Labeling Kit is designed for preparing fluorescently labeled antibodies, proteins or peptides for in vivo
imaging applications in small animals. Each kit contains our superior in vivo optimized VivoTag 680XL fluorophore and everything else youneed for carrying out the labeling reaction and purifying the labeled product.

The reactive fluorophore has a succinimidyl ester group, which reacts
with an amine group on the protein (i.e., lysine side-chain amine) toform a stable amide linkage. VivoTag 680XL is supplied in two
separate vials, each containing adequate material for labeling 0.5-5
mg of an antibody. Following the labeling reaction, the unconjugatedfluorophore is conveniently and rapidly removed by purification
columns with a molecular weight cutoff of 7 kDaCONTENTS

2 X 0.25 mg of VivoTag 680XL
1 X 1 mL of 1M solution of sodium bicarbonate (pH8.3).
2 X Purification column
4 X 15 mL conical collection tube
1 XPBS (50 mL)

PHYSICAL AND SPECTRAL PROPERTIES
Property
MW of VivoTag 680XLFluorescence Emission max1
Absorbance max1
Extinction
Purity2Appearance
1.
2.

Specification
1856 g mol-1
688 nm
668 nm210,000 M-1cm-1>95 %Blue solid

Absorbance and fluorescence maxima in 1x PBS.
As determined by SE-HPLC; measuring absorbance at 670 nm.

STORAGE & HANDLING Upon receipt, VivoTag 680XL Protein Labeling Kit should be
IMMEDIATELY STORED AT 2-8 °C AND PROTECTED FROMLIGHT.
 When stored and handled properly, contents of the labeling kits arestable for up to six months.
 Allow VivoTag 680XLto equilibrate to room temperature before
opening the vial.

Absorbance and fluorescence emission spectra in 1x PBS.

NOTES VivoTag 680XL Protein Labeling Kit is intended for research purposes only and is not for human use. It must be used by or directly under
the supervision of a technically qualified individual experienced in handling potentially hazardous materials. Please read the Material
Safety Data Sheet (MSDS) provided for this product.
 Several of PerkinElmer’s products and product applications are covered by U.S and foreign patents and patents pending. Our products
are not available for resale or other commercial uses without a specific agreement from PerkinElmer.

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LABELING PROTOCOL
1.

Prepare protein (>7 kDa) solution to 1-10 mg/mL in PBS. The protein must be free of ammonium ions or primary amines to reduce
competition for reaction with the reactive dye.

2.

Dissolve 0.25 mg of VivoTag 680XLin 10 µL of dry DMSO. Once reconstituted, VivoTag 680XL is stable for up to 7 days whenstored at 2-8 °C and protected from light.

3.

In an Eppendorf tube, add 0.5 mL of protein (0.5-5 mg), 50 µL sodium bicarbonate, 2 µL of VivoTag 680XL for each mg of protein.
Incubate in dark for 2 hours at room temperature with shaking.

4.

Separate protein conjugate from free dye. Twist off the column’s bottom closure and loosen cap. Place the column onto a 15 mL
conical collection tube and centrifuge the column at 1,000xg for 2 min. Add 2 mL of PBS to the column and centrifuge the column
at 1,000xg for 2 min. Repeat the wash two more times.

5.

Place the column to a fresh 15 mL conical collection tube. Load all the protein samples (200-700 µL) to the column and centrifuge
at 1,000xg for 2 min. Collect the flow through protein sample.

6.

The collected labeled antibody sample can be analyzed for the degree of labeling (DOL). Determine the absorbance of the purified
conjugate at 280 nm and 668 nm.

7.

Adjust the absorbance at 280 nm of the purified protein by subtracting the 280 nm absorbance of VivoTag 680XL, which is 16% ofabsorbance at 668 nm.

8.

Absorbance analysis can be done with either a UV Spectrophotometer or a Nanodrop Spectrophotometer. To use the latter,samples need to be diluted to 0.5-2 mg/mL range before measurement. As the light path is 1 mm, the reading should benormalized with a factor of 10.

DEGREE OF LABELING CALCULATIONS
Protein concentration: (M) = A280-(0.16xA668 of dye)/ ɛ (molar extinction coefficient). For antibody, ɛ is 210,000 M-1cm-1,Dye concentration (M) = A668/ ɛ (molar extinction coefficient). ɛ is 210,000 M-1cm-1 for VivoTag 680XL.
DOL (Moles dye per mole protein) = Dye concentration (M)/Protein concentration (M).
Protein recovery should be close to 100%, DOL should be between 2 and 3.

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