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iFluor® 594-Concanavalin A Conjugate

Live HeLa cells were stained with iFluor® 594-Concanavalin A Conjugate Conjugate at 5 µg/mL for 30 minutes. Image was acquired using fluorescence microscopy using TexasRed filter set.
Live HeLa cells were stained with iFluor® 594-Concanavalin A Conjugate Conjugate at 5 µg/mL for 30 minutes. Image was acquired using fluorescence microscopy using TexasRed filter set.
Live HeLa cells were stained with iFluor® 594-Concanavalin A Conjugate Conjugate at 5 µg/mL for 30 minutes. Image was acquired using fluorescence microscopy using TexasRed filter set.
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Physical properties
Molecular weightN/A
SolventWater
Spectral properties
Absorbance (nm)587
Correction Factor (260 nm)0.05
Correction Factor (280 nm)0.04
Extinction coefficient (cm -1 M -1)2000001
Excitation (nm)587
Emission (nm)603
Quantum yield0.531
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
UNSPSC12171501
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OverviewpdfSDSpdfProtocol


Molecular weight
N/A
Absorbance (nm)
587
Correction Factor (260 nm)
0.05
Correction Factor (280 nm)
0.04
Extinction coefficient (cm -1 M -1)
2000001
Excitation (nm)
587
Emission (nm)
603
Quantum yield
0.531
Concanavalin A (ConA) is a lectin that binds specifically to certain structures found in various sugars, glycoproteins and glycolipids. Con A is a well-known T cell mitogen that can activate the immune system, recruit lymphocytes and elicit cytokine production. In addition to its mitogenic activity, ConA can induce programmed cell death via mitochondria-mediated apoptosis and autophagy. ConA has also been reported to activate NFAT (nuclear factor of activated T cells), a family of transcription factors that are important in the development and function of the immune system, including T cell receptor (TCR) engagement. ConA is widely used in biology and biochemistry to characterize glycoproteins and other sugar-containing entities on the surface of various cells. It is also used to purify glycosylated macromolecules in lectin affinity chromatography, as well as to study immune regulation by various immune cells. ConA binds specifically α-D-mannosyl and α-D-glucosyl residues (two hexoses differing only in the alcohol on carbon 2) in terminal position of ramified structures from B-Glycans. It has 4 binding sites, corresponding to the 4 subunits. Concanavalin A (Con A) is one of the most widely used lectins in cell biology. iFluor®594-labeled Concanavalin A selectively binds to a-mannopyranosyl and a-glucopyranosyl residues and exhibit the bright red fluorescence.

Platform


Fluorescence microscope

ExcitationCy3/TRITC filter set
EmissionCy3/TRITC filter set
Recommended plateBlack wall/clear bottom

Example protocol


PREPARATION OF STOCK SOLUTIONS

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.

iFluor™ 594-Concanavalin A Conjugate stock solution (200X)
Add 500 µL of ddH2O into the powder form to make 2 mg/mL stock solution.
Note     The reconstituted conjugate solution can be stored at 2-8 °C for short-term storage or at -20 °C for long-term storage.

PREPARATION OF WORKING SOLUTION

iFluor™ 594-Concanavalin A Conjugate working solution (1X)
Add 5 µL of 200X Concanavalin A conjugate solution to 1 mL HHBS Buffer.
Note     The optimized staining concentration may be different with different cell lines. The recommended starting concentration is 5-10 µg/mL for live cells.

SAMPLE EXPERIMENTAL PROTOCOL

Warm the vial to room temperature centrifuge briefly before opening. Staining protocols vary with applications. Appropriate dilution of conjugates should be determined experimentally.

Live Cells Stain
  1. Wash cells twice with a HHBS buffer.
  2. Add 100 µL iFluor™ 594-Concanavalin A working solution.
  3. Incubate cells with Concanavalin A working solution for 10-30 minutes at 37 °C.
  4. Wash cells twice with HHBS buffer.
  5. Image cells on a fluorescence microscope using Cy3/TRITC filter set. 

Fixed Cells Stain
Concanavalin A conjugates can be also used to stain fixed cells.
  1. Fix cells with 4% Formaldehyde in PBS.
    Note     For fixed cell membrane staining, it is recommended to stain without permeabilization step. Permeabilized step can after fixation will lead to intracellular compartments stain such as Golgi and Endoplasmic Reticulum (ER) structures.
  2. Add 100 µL iFluor™ 594-Concanavalin A working solution.
  3. Incubate cells with Concanavalin A working solution for 10-30 minutes at room temperature.
  4. Wash cells twice with HHBS buffer.
  5. Image cells on a fluorescence microscope using Cy3/TRITC filter set. 

Spectrum


Open in Advanced Spectrum Viewer
spectrum

Spectral properties

Absorbance (nm)587
Correction Factor (260 nm)0.05
Correction Factor (280 nm)0.04
Extinction coefficient (cm -1 M -1)2000001
Excitation (nm)587
Emission (nm)603
Quantum yield0.531

Product Family


NameExcitation (nm)Emission (nm)Extinction coefficient (cm -1 M -1)Quantum yieldCorrection Factor (260 nm)Correction Factor (280 nm)
iFluor® 488-Concanavalin A Conjugate4915167500010.910.210.11
iFluor® 555-Concanavalin A Conjugate55757010000010.6410.230.14
iFluor® 647-Concanavalin A Conjugate65667025000010.2510.030.03
iFluor® 750-Concanavalin A Conjugate75777927500010.1210.0440.039

Images


References


View all 50 references: Citation Explorer
Short-term high-fat diet intake leads to exacerbation of concanavalin A-induced liver injury through the induction of procoagulation state.
Authors: Nanizawa, Eri and Tamaki, Yuki and Sono, Reika and Miyashita, Rintaro and Hayashi, Yumi and Kanbe, Ayumu and Ito, Hiroyasu and Ishikawa, Tetsuya
Journal: Biochemistry and biophysics reports (2020): 100736
Maresin 1 mitigates concanavalin A-induced acute liver injury in mice by inhibiting ROS-mediated activation of NF-κB signaling.
Authors: Zhang, Peng and Yin, Yuping and Wang, Tao and Li, Wei and Li, Chengguo and Zeng, Xiangyu and Yang, Wenchang and Zhang, Ruizhi and Tang, Yu and Shi, Liang and Li, Ruidong and Tao, Kaixiong
Journal: Free radical biology & medicine (2020): 23-36
Membrane-associated gamma-glutamyl transferase and alkaline phosphatase in the context of concanavalin A- and wheat germ agglutinin-reactive glycans mark seminal prostasome populations from normozoospermic and oligozoospermic men.
Authors: Janković, Tamara and Goč, Sanja and Mitić, Ninoslav and Danilović Luković, Jelena and Janković, Miroslava
Journal: Upsala journal of medical sciences (2020): 10-18
Antioxidant Capacity and Hepatoprotective Role of Chitosan-Stabilized Selenium Nanoparticles in Concanavalin A-Induced Liver Injury in Mice.
Authors: Bai, Kaikai and Hong, Bihong and He, Jianlin and Huang, Wenwen
Journal: Nutrients (2020)
The farnesyltransferase inhibitor tipifarnib protects against autoimmune hepatitis induced by Concanavalin A.
Authors: Guo, Jie and Shirozu, Kazuhiro and Akahoshi, Tomohiko and Mizuta, Yukie and Murata, Masaharu and Yamaura, Ken
Journal: International immunopharmacology (2020): 106462
Concanavalin A-Rose Bengal bioconjugate for targeted Gram-negative antimicrobial photodynamic therapy.
Authors: Cantelli, Andrea and Piro, Francesca and Pecchini, Pietro and Di Giosia, Matteo and Danielli, Alberto and Calvaresi, Matteo
Journal: Journal of photochemistry and photobiology. B, Biology (2020): 111852
Developmental exposure to low doses of dichlorodiphenyltrichloroethane impairs proliferative response of thymic lymphocytes to Concanavalin A in rats.
Authors: Yaglova, Nataliya V and Tsomartova, Elina S and Obernikhin, Sergey S and Ivanova, Marina Y and Chereshneva, Elizaveta V and Muhamedova, Svetlana G and Lomanovskaya, Tatiana A and Yaglov, Valentin V
Journal: Heliyon (2020): e03608
Concanavalin A Toxicity Towards Potato Psyllid and Apoptosis Induction in Midgut Cells.
Authors: Tang, Xiao-Tian and Ibanez, Freddy and Tamborindeguy, Cecilia
Journal: Insects (2020)
Concanavalin A Enhanced Proliferation and Osteogenic Differentiation of Dental Pulp Stem Cells.
Authors: Suardita, Ketut and Arundina, Ira and Tedjosasongko, Udijanto and Yuliati, Anita and Peeters, Harry Huiz and Wijaksana, I Komang Evan and Surboyo, Meircurius Dwi Condro
Journal: European journal of dentistry (2020): 123-127
Mycophenolate mofetil attenuates concanavalin A-induced acute liver injury through modulation of TLR4/NF-κB and Nrf2/HO-1 pathways.
Authors: Serrya, Marwa S and Zaghloul, Marwa S
Journal: Pharmacological reports : PR (2020)