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mFluor™ Violet 530 SE

Ordering information
Price ()
Catalog Number1616
Unit Size
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Additional ordering information
InternationalSee distributors
ShippingStandard overnight for United States, inquire for international
Physical properties
Molecular weight1174.17
Spectral properties
Absorbance (nm)398
Extinction coefficient (cm -1 M -1)200001
Excitation (nm)393
Emission (nm)543
Storage, safety and handling
H-phraseH303, H313, H333
Hazard symbolXN
Intended useResearch Use Only (RUO)
R-phraseR20, R21, R22
StorageFreeze (< -15 °C); Minimize light exposure


Molecular weight
Absorbance (nm)
Extinction coefficient (cm -1 M -1)
Excitation (nm)
Emission (nm)
mFluor™ Violet 530 dyes have fluorescence excitation and emission maxima of ~405 nm and ~530 nm respectively. These spectral characteristics make them a unique color for flow cytometry application. mFluor™ Violet 530 SE is reasonably stable and shows good reactivity and selectivity with protein amino groups. It provides a convenient tool to label monoclonal, polyclonal antibodies or other proteins (>10 kDa) for flow cytometric applications with the violet laser excitation, in particular suitable for spectral flow cytometric applications. mFluor™ dyes are developed for multicolor flow cytometry-focused applications. These dyes have large Stokes Shifts, and can be well excited by the laser lines of flow cytometers (e.g., 350 nm, 405 nm, 488 nm and 633 nm). mFluor™ Violet dyes are optimized to be excited with a Violet laser at 405 nm. AAT Bioquest offers the largest collection of fluorescent dyes that are excited by Violet laser at 405 nm.

Example protocol


Unless otherwise noted, all unused stock solutions should be divided into single-use aliquots and stored at -20 °C after preparation. Avoid repeated freeze-thaw cycles.

1. Protein stock solution (Solution A)
Mix 100 µL of a reaction buffer (e.g., 1 M  sodium carbonate solution or 1 M phosphate buffer with pH ~9.0) with 900 µL of the target protein solution (e.g. antibody, protein concentration >2 mg/mL if possible) to give 1 mL protein labeling stock solution. Note: The pH of the protein solution (Solution A) should be 8.5 ± 0.5. If the pH of the protein solution is lower than 8.0, adjust the pH to the range of 8.0-9.0 using 1 M  sodium bicarbonate solution or 1 M pH 9.0 phosphate buffer. Note: The protein should be dissolved in 1X phosphate buffered saline (PBS), pH 7.2-7.4. If the protein is dissolved in Tris or glycine buffer, it must be dialyzed against 1X PBS, pH 7.2-7.4, to remove free amines or ammonium salts (such as ammonium sulfate and ammonium acetate) that are widely used for protein precipitation. Note: Impure antibodies or antibodies stabilized with bovine serum albumin (BSA) or gelatin will not be labeled well. The presence of sodium azide or thimerosal might also interfere with the conjugation reaction. Sodium azide or thimerosal can be removed by dialysis or spin column for optimal labeling results. Note: The conjugation efficiency is significantly reduced if the protein concentration is less than 2 mg/mL. For optimal labeling efficiency the final protein concentration range of 2-10 mg/mL is recommended.

2. mFluor™ Violet 530 SE stock solution (Solution B)
Add anhydrous DMSO into the vial of mFluor™ Violet 530 SE to make a 10 mM stock solution. Mix well by pipetting or vortex. Note: Prepare the dye stock solution (Solution B) before starting the conjugation. Use promptly. Extended storage of the dye stock solution may reduce the dye activity. Solution B can be stored in freezer for two weeks when kept from light and moisture. Avoid freeze-thaw cycles.


This labeling protocol was developed for the conjugate of Goat anti-mouse IgG with mFluor™ Violet 530 SE. You might need further optimization for your particular proteins. Note: Each protein requires distinct dye/protein ratio, which also depends on the properties of dyes. Over labeling of a protein could detrimentally affects its binding affinity while the protein conjugates of low dye/protein ratio gives reduced sensitivity.

Run conjugation reaction
  1. Use 10:1 molar ratio of Solution B (dye)/Solution A (protein) as the starting point:  Add 5 µL of the dye stock solution (Solution B, assuming the dye stock solution is 10 mM) into the vial of the protein solution (95 µL of Solution A) with effective shaking. The concentration of the protein is ~0.05 mM assuming the protein concentration is 10 mg/mL and the molecular weight of the protein is ~200KD. Note: We recommend to use 10:1 molar ratio of Solution B (dye)/Solution A (protein). If it is too less or too high, determine the optimal dye/protein ratio at 5:1, 15:1 and 20:1 respectively.
  2. Continue to rotate or shake the reaction mixture at room temperature for 30-60 minutes. 

Purify the conjugation
The following protocol is an example of dye-protein conjugate purification by using a Sephadex G-25 column.
  1. Prepare Sephadex G-25 column according to the manufacture instruction.
  2. Load the reaction mixture (From "Run conjugation reaction") to the top of the Sephadex G-25 column.
  3. Add PBS (pH 7.2-7.4) as soon as the sample runs just below the top resin surface.
  4. Add more PBS (pH 7.2-7.4) to the desired sample to complete the column purification. Combine the fractions that contain the desired dye-protein conjugate. Note: For immediate use, the dye-protein conjugate need be diluted with staining buffer, and aliquoted for multiple uses. Note: For longer term storage, dye-protein conjugate solution need be concentrated or freeze dried. 


Common stock solution preparation

Table 1. Volume of DMSO needed to reconstitute specific mass of mFluor™ Violet 530 SE to given concentration. Note that volume is only for preparing stock solution. Refer to sample experimental protocol for appropriate experimental/physiological buffers.

0.1 mg0.5 mg1 mg5 mg10 mg
1 mM85.167 µL425.833 µL851.665 µL4.258 mL8.517 mL
5 mM17.033 µL85.167 µL170.333 µL851.665 µL1.703 mL
10 mM8.517 µL42.583 µL85.167 µL425.833 µL851.665 µL

Molarity calculator

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Spectral properties

Absorbance (nm)398
Extinction coefficient (cm -1 M -1)200001
Excitation (nm)393
Emission (nm)543

Product family

NameExcitation (nm)Emission (nm)Extinction coefficient (cm -1 M -1)Quantum yieldCorrection Factor (260 nm)Correction Factor (280 nm)
mFluor™ Violet 450 SE4064453500010.8110.3380.078
mFluor™ Violet 510 SE4125052500010.8610.4640.366
mFluor™ Violet 540 SE4025351800010.2111.3260.543
mFluor™ Violet 500 SE4105012500010.8110.7690.365
mFluor™ Violet 610 SE4216133500010.310.5320.66
mFluor™ Violet 550 SE5275509000010.3110.4740.306
mFluor™ Violet 505 SE3935044000010.4510.8880.403
mFluor™ Violet 590 SE5645919000010.2210.6320.329
mFluor™ Violet 545 SE3935432000010.1511.080.496
mFluor™ Violet 530 maleimide393543200001---
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