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VRDye 490 Secondary Antibodies

Products

VRDye 490 secondary antibodies can be used for a variety of applications, including flow cytometry, immunofluorescence microscopy, and others.

These secondary antibodies are suitable for multiplex experiments when combined with other secondary antibodies labeled with proper fluorescent dyes and using instrumentation with appropriate excitation and detection capabilities.

Visible Fluorescence Reagents Overview

VRDye 490 Forms Available

  • Secondary antibodies
  • Labeling kits

Formulation and Recommended Dilutions

VRDye 490 secondary antibodies are supplied as purified immunoglobulin conjugates, lyophilized in phosphate-buffered saline, pH 7.4. Protect from light. Store at 4 °C prior to reconstitution.

Each vial contains 10 mg/mL BSA (free of IgG and protease) as a stabilizer and 0.01% sodium azide as a preservative, after reconstitution. Concentration is 1.0 mg/mL when reconstituted as directed. Refer to the pack insert for details on reconstitution.

Application Recommended Suggested Range
Immunofluorescence Microscopy 1:400 1:100 - 1:1,000
Flow Cytometry 1:1,000 1:200 - 1:2,000
Other User optimized

Suggested working dilutions are given as a guide only. Optimum dilutions will vary and should be determined empirically.

Absorption and Emission Spectra in 1X PBS

Properties of Visible Fluorescence NHS Ester Dyes

Dye Absmax Emmax ε(M-1cm-1) MW (g/mole) CF*
VRDye 490 491 515 73,000 1,011 0.11

*CF is a correction factor for the absorbance of the dye at Absmax to the absorbance of the protein at 280 nm.

VRDye 490 Used for Immunofluorescence

Immunofluorescence staining of tubulin protein in HeLa cells. Cells were cultured on cover slips. After fixation and permeabilization, cells were incubated with rabbit anti-tubulin mAb (CST), followed by VRDye 490 Goat anti-Rabbit IgG (P/N 926-49020). Nuclei were stained with DAPI. Image acquired with Olympus IX81 microscope.
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