Cy5 Dyes – VectorLabs

Cy5 Amine

Vector Laboratories R&D — Tue, 19 Sep 2023 22:34:54 +0000

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Description

Cy5 Amine is a carbonyl reactive building block used to modify carboxylic groups in the presence of activators (e.g. EDC, or DCC) or activated esters (e.g. NHS esters) through a stable amide bond. It also can be employed as a polar tracer and as a reactive dye for labeling proteins via a carboxylic acid moiety.

Cy5 Amine (also sold under the Sulfo-Cyanine5 Amine name) is a water-soluble, bright, far-red-fluorescent probe with excitation ideally suited for the 633 nm or 647 nm laser lines. A significant advantage to using long wavelength dyes such as Cy5 or AZDye 647 over other fluorophores is the low autofluorescence of biological specimens in this region of the spectrum.

Specifications

Unit Size	1 mg, 5 mg, 25 mg, 100 mg
Reactivity	Aldehyde, carboxylic acid, ketone
Abs/Em Maxima	648/671 nm
Extinction coefficient	250,000 cm-1M-1
Solubility	Water, DMSO, DMF
Spectrally similar dyes	Alexa Fluor® 647, DyLight® 649,
Molecular weight	698.98
Storage Conditions	-20°C.
Shipping Conditions	Ambient temperature

Abs/Em Spectra

Protein Labeling Calculator

Protein name

Protein molecular weight (daltons)

Protein concentration (mg/mL)

Volume of protein to be labeled (µL)

Enter only one — either volume or amount of protein to be labeled

Amount of protein to be labeled (mg)

Molecular weight of labeling reagent (Da)

Amount of labeling reagent used (mg)

Maximum concentration of DMSO (%) Leave blank or 0 if not used

Enter only one — either maximum concentration of DMSO or volume DMSO added to label

Volume of anhydrous DMSO added (µL)

Molar equivalents of labeling reagent used

Volume of DMSO required to make a stock solution (µL)

Volume of protein to be labeled (µL)

Amount of stock labeling reagent to be added (µL)

Amount of DMSO in labeling reaction (%)

Documents

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Cy5 Maleimide

Vector Laboratories R&D — Tue, 19 Sep 2023 22:34:29 +0000

Description
Specifications
Abs/Em Spectra
Protein Labeling Calculator
Documents

Description

Cy5 Maleimide (also sold under Sulfo-Cyanine5 Maleimide) is a water-soluble, bright, far-red-fluorescent dye with excitation ideally suited for the 633 nm or 647 nm laser lines. A significant advantage to using long wavelength dyes such as Cy5 or AF 647 dye over other fluorophores is the low autofluorescence of biological specimens in this region of the spectrum.

Maleimide is the most popular sulfhydryl-reactive reactive group for conjugating the dye to a thiol group on a protein, oligonucleotide thiophosphate, or low molecular weight ligand. We do not recommended using Cy5 Maleimide for labeling proteins at high molar ration due significant self-quenching. Cy5 conjugates are only recommended for detection of moderate-to-high abundance targets. For detection of low-abundance biological targets we recommend using AF 647 Maleimide (Alexa Fluor ® 647 analog) or IR 647 (IRDye® 647 analog), which can attached to proteins at high molar ratios without significant self-quenching, enabling brighter conjugates and more sensitive detection.

Specifications

Unit Size	1 mg, 5 mg, 25 mg, 100 mg
Reactivity	Thiol (Cys)
Abs/Em Maxima	648/671 nm
Extinction coefficient	250,000 cm-1M-1
Solubility	Water, DMSO, DMF
Spectrally similar dyes	Alexa Fluor® 647, DyLight® 649,
Molecular weight	764.91
Storage Conditions	-20°C.
Shipping Conditions	Ambient temperature

Abs/Em Spectra

Protein Labeling Calculator

Protein name

Protein molecular weight (daltons)

Protein concentration (mg/mL)

Volume of protein to be labeled (µL)

Enter only one — either volume or amount of protein to be labeled

Amount of protein to be labeled (mg)

Molecular weight of labeling reagent (Da)

Amount of labeling reagent used (mg)

Maximum concentration of DMSO (%) Leave blank or 0 if not used

Enter only one — either maximum concentration of DMSO or volume DMSO added to label

Volume of anhydrous DMSO added (µL)

Molar equivalents of labeling reagent used

Volume of DMSO required to make a stock solution (µL)

Volume of protein to be labeled (µL)

Amount of stock labeling reagent to be added (µL)

Amount of DMSO in labeling reaction (%)

Documents

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Sulfo Cy5 Maleimide

Vector Laboratories R&D — Tue, 19 Sep 2023 22:34:27 +0000

Description
Specifications
Abs/Em Spectra
Protein Labeling Calculator
Documents

Description

Sulfo Cy5 Maleimide is a water-soluble, bright, far-red-fluorescent dye with excitation ideally suited for the 633 nm or 647 nm laser lines. A significant advantage to using long wavelength dyes such as Sulfo Cy5, AZDye 647 or IR 650 dye over other fluorophores is the low autofluorescence of biological specimens in this region of the spectrum.

Maleimide is the most popular sulfhydryl-reactive reactive group for conjugating the dye to a thiol group on a protein, oligonucleotide thiophosphate, or low molecular weight ligand. Sulfo Cy5 conjugates are only recommended for detection of moderate-to-high abundance targets. For detection of low-abundance biological targets we recommend using AZDye 647 Maleimide or IR 647 Maleimide, which can attached to proteins at high molar ratios without significant self-quenching, enabling brighter conjugates and more sensitive detection.

Specifications

Unit Size	1 mg, 5 mg, 25 mg, 100 mg
Reactivity	Thiol (Cys)
Abs/Em Maxima	648/671 nm
Extinction coefficient	250,000 cm-1M-1
Solubility	Water, DMSO, DMF
Spectrally similar dyes	Alexa Fluor® 647, DyLight® 649,
Molecular weight	872.26
Storage Conditions	-20°C.
Shipping Conditions	Ambient temperature

Abs/Em Spectra

Protein Labeling Calculator

Protein name

Protein molecular weight (daltons)

Protein concentration (mg/mL)

Volume of protein to be labeled (µL)

Enter only one — either volume or amount of protein to be labeled

Amount of protein to be labeled (mg)

Molecular weight of labeling reagent (Da)

Amount of labeling reagent used (mg)

Maximum concentration of DMSO (%) Leave blank or 0 if not used

Enter only one — either maximum concentration of DMSO or volume DMSO added to label

Volume of anhydrous DMSO added (µL)

Molar equivalents of labeling reagent used

Volume of DMSO required to make a stock solution (µL)

Volume of protein to be labeled (µL)

Amount of stock labeling reagent to be added (µL)

Amount of DMSO in labeling reaction (%)

Documents

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Cy5 NHS ester

Vector Laboratories R&D — Tue, 19 Sep 2023 22:33:38 +0000

Description
Specifications
Abs/Em Spectra
Protein Labeling Calculator
Documents

Description

Cy5 dye (also sold under Sulfo-Cyanine5 name) is one of the most popular far-red fluorescent dyes. It is often a reagent of choice for protein and peptide labeling. Cy5 dye is a water-soluble, bright, far-red-fluorescent dye with excitation ideally suited for the 633 nm or 647 nm laser lines. Cy5 conjugates of antibodies, peptides, and proteins are pH insensitive from pH 4 to pH 10. A significant advantage to using long wavelength dyes such as Cy5 or AF 647 dye over other fluorophores is the low autofluorescence of biological specimens in this region of the spectrum.

The NHS ester (or succinimidyl ester) is the most popular amine reactive group for labeling with the primary amines of proteins (Lys), amine-modified oligonucleotides, and other amine-containing molecules. We do not recommend using Cy5 NHS ester for labeling proteins at high molar ration due significant self-quenching. Cy5 conjugates are recommended for detection of moderate-to-high abundance targets. For detection of low-abundance biological targets we recommend using AZDye 647 (Alexa Fluor® 647 analog), or IR 650 dye (IRDye® 650 analog), which can be attached to proteins at high molar ratios without significant self-quenching, enabling brighter conjugates and more sensitive detection.

Specifications

Unit Size	1 mg, 5 mg, 25 mg, 100 mg
Reactivity	Primary amine
Abs/Em Maxima	648/671 nm
Extinction coefficient	250,000 cm-1M-1
Solubility	Water, DMSO, DMF
Spectrally similar dyes	Alexa Fluor® 647, DyLight® 649,
Molecular weight	739.86
Storage Conditions	-20°C.
Shipping Conditions	Ambient temperature

Abs/Em Spectra

Protein Labeling Calculator

Protein name

Protein molecular weight (daltons)

Protein concentration (mg/mL)

Volume of protein to be labeled (µL)

Enter only one — either volume or amount of protein to be labeled

Amount of protein to be labeled (mg)

Molecular weight of labeling reagent (Da)

Amount of labeling reagent used (mg)

Maximum concentration of DMSO (%) Leave blank or 0 if not used

Enter only one — either maximum concentration of DMSO or volume DMSO added to label

Volume of anhydrous DMSO added (µL)

Molar equivalents of labeling reagent used

Volume of DMSO required to make a stock solution (µL)

Volume of protein to be labeled (µL)

Amount of stock labeling reagent to be added (µL)

Amount of DMSO in labeling reaction (%)

Documents

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Sulfo-Cy5 NHS ester

Vector Laboratories R&D — Tue, 19 Sep 2023 22:33:36 +0000

Description
Specifications
Abs/Em Spectra
Protein Labeling Calculator
Documents

Description

Sulfo-Cy5 dye is one of the most popular far-red fluorescent dyes. It is often a reagent of choice for protein and peptide labeling. Sulfo-Cy5 dye is a water-soluble, bright, far-red-fluorescent dye with excitation ideally suited for the 633 nm or 647 nm laser lines. Sulfo-Cy5 conjugates of antibodies, peptides, and proteins are pH insensitive from pH 4 to pH 10. A significant advantage to using long wavelength dyes such as Cy5 or AF 647 dye over other fluorophores is the low autofluorescence of biological specimens in this region of the spectrum.

The NHS ester (or succinimidyl ester) is the most popular amine reactive group for labeling with the primary amines of proteins (Lys), amine-modified oligonucleotides, and other amine-containing molecules. Sulfo-Cy5 conjugates are recommended for detection of moderate-to-high abundance targets. For detection of low-abundance biological targets we recommend using AZDye 647 NHS Ester analog, or IR 650 NHS Ester, which can be attached to proteins at high molar ratios without significant self-quenching, enabling brighter conjugates and more sensitive detection.

Specifications

Unit Size	1 mg, 5 mg, 25 mg, 100 mg
Reactivity	Primary amine
Abs/Em Maxima	648/671 nm
Extinction coefficient	250,000 cm-1M-1
Solubility	Water, DMSO, DMF
Spectrally similar dyes	Alexa Fluor® 647, DyLight® 649,
Molecular weight	847.97
Storage Conditions	-20°C.
Shipping Conditions	Ambient temperature

Abs/Em Spectra

Protein Labeling Calculator

Protein name

Protein molecular weight (daltons)

Protein concentration (mg/mL)

Volume of protein to be labeled (µL)

Enter only one — either volume or amount of protein to be labeled

Amount of protein to be labeled (mg)

Molecular weight of labeling reagent (Da)

Amount of labeling reagent used (mg)

Maximum concentration of DMSO (%) Leave blank or 0 if not used

Enter only one — either maximum concentration of DMSO or volume DMSO added to label

Volume of anhydrous DMSO added (µL)

Molar equivalents of labeling reagent used

Volume of DMSO required to make a stock solution (µL)

Volume of protein to be labeled (µL)

Amount of stock labeling reagent to be added (µL)

Amount of DMSO in labeling reaction (%)

Documents

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Sulfo-Cy5 Acid

Vector Laboratories R&D — Tue, 19 Sep 2023 22:33:10 +0000

Description
Specifications
Abs/Em Spectra
Documents

Description

The carboxylic acid of Sulfo Cy5 dye is a reagent of choice for the preparation of custom activated esters that often are not commercially available. Examples of such activated esters include sulfo-NHS, TFP (2,3,5,6-Tetrafluorophenol), STP (4-Sulfo-2,3,5,6-Tetrafluorophenol, Sodium Salt). Another common application for non-activated carboxylic acid is peptide modification during solid phase synthesis, which usually requires in-situ activation with peptide coupling regents, for example HATU. Cy5 Acid is also often used for control experiments, and for calibration.

Specifications

Unit Size	5 mg, 25 mg, 100 mg
Reactivity	Primary amine
Abs/Em Maxima	648/671 nm
Extinction coefficient	250,000 cm-1M-1
Solubility	Water, DMSO, DMF
Spectrally similar dyes	Alexa Fluor® 647, DyLight® 649,
Molecular weight	750.89
Storage Conditions	-20°C.
Shipping Conditions	Ambient temperature

Abs/Em Spectra

Documents

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Cy5 Azide NHS Ester

Vector Laboratories R&D — Tue, 19 Sep 2023 20:22:52 +0000

Description
Specifications
Abs/Em Spectra
Documents

Description

Cy5 Azide NHS Ester is a bright, far red fluorescent dual-label click chemistry probe that allows for simultaneous attachment of azide group and far red fluorescent Cy5 label. Conjugates prepared with Cy5 Azide NHS Ester can be reacted with terminal alkynes via a copper-catalyzed click reaction (CuAAC). It also reacts with strained cyclooctyne via a copper-free “click chemistry” reaction to form a stable triazole and does not require Cu-catalyst or elevated temperatures.

Cy5 dye is a water-soluble, pH-insensitive from pH 4 to pH 10, far-red-fluorescent probe with excitation ideally suited for the 633 nm or 647 nm laser lines. Its absorption and emission spactra are almost identical to those of Alexa Fluor® 647, CF® 647 Dye, or any other Cyanine5 based fluorescent dyes.

Specifications

Unit Size	1 mg, 5 mg
Abs/Em Maxima	649/671 nm
Extinction Coefficient	250,000
Flow Cytometry Laser Line	633 or 635 nm
Microscopy Laser Line	633 or 635 nm
Spectrally Similar Dyes	Alexa Fluor® 647, CF™ 647 Dye, DyLight™649
Molecular weight	922.08
CAS	1564286-24-3
Solubility	Water, DMSO, DMF
Appearance	Blue solid
Storage Conditions	-20°C. Desiccate
Shipping Conditions	Ambient temperature

Abs/Em Spectra

Documents

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Cy5 DBCO NHS Ester

Vector Laboratories R&D — Tue, 19 Sep 2023 20:18:00 +0000

Description
Specifications
Abs/Em Spectra
Documents
Selected References

Description

Cy5 DBCO NHS Ester is a bright, far red fluorescent dual-label click chemistry probe that allows for simultaneous attachment of DBCO moiety and far red fluorescent Cy5 label. Conjugates prepared with Cy5 DBCO NHS Ester reacts with azides via a copper-free “click chemistry” reaction to form a stable triazole and does not require Cu-catalyst or elevated temperatures. In application where the presence of copper is a concern Cy5 DBCO is an ideal alternative to copper requiring fluorescent alkynes.

Specifications

Unit Size	1 mg, 5 mg
Abs/Em Maxima	649/671 nm
Extinction Coefficient	250,000
Flow Cytometry Laser Line	633 or 635 nm
Microscopy Laser Line	633 or 635 nm
Spectrally Similar Dyes	Alexa Fluor® 647, CF™ 647 Dye, DyLight™649
Molecular weight	1098.30
CAS	N/A
Solubility	Water, DMSO, DMF
Appearance	Blue solid
Storage Conditions	-20°C. Desiccate
Shipping Conditions	Ambient temperature

Abs/Em Spectra

Documents

Selected References

Bazrafshan, A. et al. (2021). DNA Gold Nanoparticle Motors Demonstrate Processive Motion with Bursts of Speed Up to 50 nm Per Second. ACS Publications, Online ahead of print. [PubMed]
Wiener, J., et al. (2020). Preparation of single- and double-oligonucleotide antibody conjugates and their application for protein analytics. Sci Rep., 10 (1), 1457. [PubMed]
Yong, K.W., et al. (2020). Engineering the Orientation, Density, and Flexibility of Single-Domain Antibodies on Nanoparticles To Improve Cell Targeting. ACS Appl. Mater. Interfaces, 12(5), 5593-600. [PubMed]
Song, S., et al. (2020). In Situ One-Step Fluorescence Labeling Strategy of Exosomes via Bioorthogonal Click Chemistry for Real-Time Exosome Tracking In Vitro and In Vivo. Bioconjugate Chem., 31(5), 1562-74. [PubMed]
Valentini, T.V., et al. (2020). Bioorthogonal non-canonical amino acid tagging reveals translationally active subpopulations of the cystic fibrosis lung microbiota. Nat Commun., 11, 2287-74. [PMC]
Kim, S. H., et al. (2014). Cell labeling and tracking method without distorted signals by phagocytosis of macrophages . Theranostics, 4 (4), 420-31. [PubMed]

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Cy5 Tyramide

Vector Laboratories R&D — Tue, 19 Sep 2023 20:11:03 +0000

Description
Specifications
Abs/Em Spectra
Documents
FAQs

Description

Far-red fluorescent reagent widely used for tyramide signal amplification (TSA) in IHC, ICC, FISH and multicolor FISH. Tyramide signal amplification (TSA) has proven to be a particularly versatile and powerful enzyme amplification technique with improved assay sensitivity. TSA is based on the ability of HRP, in the presence of low concentrations of hydrogen peroxide, to convert labeled tyramine-containing substrate into an oxidized, highly reactive free radical that can covalently bind to tyrosine residues at or near the HRP.

Cy5 Tyramide is a component of many tyramide signal amplification kits. This reagent can be used with any antibody conjugated to HRP for tyramide signal amplification for detection of low abundance targets in multiplexable fluorescent immunocytochemistry (ICC), immunohistochemistry (IHC ), and in situ hybridization (ISH).

Specifications

Unit Size	150 slides, 1500 slides
Abs/Em Maxima	648/671 nm
Extinction Coefficient	270,000
Flow Cytometry Laser Line	633 nm or 647 nm
Microscopy Laser Line	633 nm or 647 nm
Spectrally Similar Dyes	Alexa Fluor® 647, CF® 647, DyLight® 649
Molecular weight	761.28 (protonated)
CAS	N/A
Solubility	Water, DMSO, DMF
Appearance	Blue solid
Storage Conditions	-20°C. Desiccate
Shipping Conditions	Ambient temperature

Abs/Em Spectra

Documents

FAQs

What is the quantity of tyramide reagent provided in the 150 slides and 1500 slides unit sizes?

150 slides has 0.24 µmol (depending on molecular weight this is ~200 µg)

1500 slides has 2.4 µmol (depending on molecular weight this is ~2 mg)

What is the protocol for using a Fluorophore Tyramide product in tyramide signal amplification (TSA)?

Here is a general protocol for the 150 slide unit size. To prepare 100x stock solution, we recommend reconstituting with 150 ul of DMSO. Storage once in solution: 100X stock solution in DMSO can be stored for 6 months at 2-8 °C protected from light. Avoid moisture.

Working solutions can be made fresh by adding 1 part 100X stock solution, 1 part 0.15% H202, and 100 part buffer.

For example, you can add 5 µL of 100X Tyramide stock solution and 5 µL 0.15% H2O2 to 500 µL of buffer. The final concentration of H2O2 is 0.0015%.

Typical reaction time following the HRP reagent is 2-10 minutes but should be optimized for each workflow. Following the TSA working solution incubation, wash thoroughly.

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Cy5 Azide Plus

Vector Laboratories R&D — Tue, 19 Sep 2023 19:59:30 +0000

Description
Specifications
Abs/Em Spectra
Documents
Click Reaction Protocol

Description

Azide Plus reagents is the most recent step in improving CuAAC reaction in complex media developed by scientists at Click Chemistry Tools. Azide Plus reagents contain a complete copper-chelating system in their structure, allowing for the formation of strong, active copper complexes that act simultaneously as both reactant and catalyst in the CuAAC reaction. This azide-copper complex reacts almost instantaneously with alkynes under diluted conditions. This unprecedented reactivity in the CuAAC reaction is of special value for the detection of low abundance targets, improving biocompatibility, and is also valuable for any other application where greatly improved S/N ratio is highly desired.

Specifications

Unit Size	1 mg, 5 mg, 25 mg
Abs/Em Maxima	647/663 nm
Extinction Coefficient	250,000
Flow Cytometry Laser Line	633 nm or 647 nm
Microscopy Laser Line	633 nm or 647 nm
Spectrally Similar Dyes	Alexa Fluor® 647, CF® 647, DyLight® 649
Molecular weight	971.18 (protonated)
CAS	N/A
Solubility	Water, DMSO, DMF
Purity	>95% (HPLC)
Appearance	Blue solid
Storage Conditions	-20°C. Desiccate
Shipping Conditions	Ambient temperature

Abs/Em Spectra

Documents

Click Reaction Protocol for Staining Fixed/Permeabilized Cell

This is a general protocol for fixed/permeabilized cell imaging through a copper-catalyzed click reaction using the fluorescent Azide Plus reagent. We recommend using this protocol as a starting point for optimization of particular click chemistry procedures. We have found that a 1.5-3.0 μM concentration of Azide Plus reagent was optimal for most applications, including imaging of EdU incorporated into newly synthesized DNA and imaging of OPP labeled proteins without causing a high background signal. The optimal final concentration of the Azide Plus reagent is sample dependent and may range from 0.5 μM to 10 μM. Final concentrations below or above this range are also possible, and should be optimized per the specific application.

Prepare the following click solutions:
— 50 mM copper sulfate in water
— 300 mM sodium ascorbate in water (dissolve 60 mg of sodium ascorbate in 1 mL of water)
— Subtext

1 mM Azide Plus reagent in DMSO or water

Table 1

Reaction Component	Number of coverslipsor wells of a 96-well plate
Reaction Component	1 coverslip or 10 wells	5 coverslips or 50 wells	10 coverslips or 100 wells	20 coverslips or 200 wells
Reaction Buffer (Tris)	888 µL	4.4 mL	8.9 mL	17.8 mL
50 mM Copper Sulfate	10 µL	50 µL	100 µL	200 µL
AZDye Azide Plus Solution (2 µM final concentration)	2 µL	10 µL	20 µL	40 µL
Sodium ascorbate	100 µL	500 µL	1 mL	2 mL
Total Volume	1 mL	5 mL	10 mL	20 mL

Remove the permeabilization buffer (if used). Wash the cells in each well twice with 1 ml of PBS. Remove PBS.
Immediately add 1 mL of the Reaction Cocktail to the sample. Evenly distribute the reaction cocktail over the sample.
Protect from light, and incubate the plate for 30 minutes at room temperature.
6Remove the reaction cocktail. Wash each well once with 1 ml of Wash Buffer. Remove the Wash Buffer.
Wash each well with 1 mL of PBS. Remove PBS.

Click Reaction Protocol for Cell Lysates Labeling

This is a general protocol for labeling proteins in cell lysate through a copper-catalyzed click reaction using the fluorescent Azide Plus reagent. We recommend using this protocol as a starting point for optimization of particular click chemistry procedures. We have found that a 20 μM concentration of Azide Plus reagent was sufficient to label all alkyne-tagged proteins in the cell lysate without causing a high background signal. The optimal final concentration of the Azide Plus reagent is sample dependent and may range from 5 μM to 50 μM. Final concentrations below or above this range are also possible, and should be optimized per the specific application.

Prepare the following click solutions:
— 100 mM THPTA ligand in water (100 mg of THPTA in 2.3 mL of water)
— 20 mM copper sulfate in water (dissolve 11.6 mg of copper II sulfate pentahydrate in 2.3 mL of water)
— 300 mM sodium ascorbate in water (dissolve 60 mg of sodium ascorbate in 1 mL of water)
— 1 mM Azide Plus reagent in DMSO or water
For each protein lysate sample, add the following to a 1.5 mL microfuge tube, then vortex briefly to mix.
— 50 µL of protein lysate (1-5 mg/mL) in protein extraction buffer
— 120 µL of Tris buffer
— 4 µL of Azide Plus reagent stock solution (5 μM final concentration)
Add 10 µL of 100 mM THPTA solution, vortex briefly to mix.
Add 10 µL of 20 mM CuSO₄ solution, vortex briefly to mix.
Add 10 µL of 300 mM sodium ascorbate solution to initiate click reaction, vortex briefly to mix.
Vortex continuously or rotate end-over-end for 30 minutes at room temperature.
Add the labeling reaction to 3 mL of cold (–20°C) methanol, 0.75 ml of Chloroform and 2.1 mL of water. Cool it to –20°C for 1 hour.
Centrifuge for 10 minutes at 13,000-20,000 x g, then carefully remove upper aqueous layer without disturbing the interface layer containing proteins.
Add 450 µL of methanol, vortex briefly.
Centrifuge for 5 minutes at 13,000-20,000 × g to pellet protein. Carefully remove and discard supernatant.
Open the lid to microfuge tube and allow protein pellet to air dry. Do not over dry the pellet!

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