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iFluor® 488-Concanavalin A Conjugate

Live HeLa cells were stained with iFluor® 488-Concanavalin A Conjugate Conjugate at 5 µg/mL for 30 minutes. Image was acquired using fluorescence microscopy using FITC filter set.
Live HeLa cells were stained with iFluor® 488-Concanavalin A Conjugate Conjugate at 5 µg/mL for 30 minutes. Image was acquired using fluorescence microscopy using FITC filter set.
Live HeLa cells were stained with iFluor® 488-Concanavalin A Conjugate Conjugate at 5 µg/mL for 30 minutes. Image was acquired using fluorescence microscopy using FITC filter set.
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Physical properties
Molecular weightN/A
SolventWater
Spectral properties
Correction Factor (260 nm)0.21
Correction Factor (280 nm)0.11
Extinction coefficient (cm -1 M -1)750001
Excitation (nm)491
Emission (nm)516
Quantum yield0.91
Storage, safety and handling
Certificate of OriginDownload PDF
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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Show More (277)

OverviewpdfSDSpdfProtocol


Molecular weight
N/A
Correction Factor (260 nm)
0.21
Correction Factor (280 nm)
0.11
Extinction coefficient (cm -1 M -1)
750001
Excitation (nm)
491
Emission (nm)
516
Quantum yield
0.91
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®488-labeled Concanavalin A selectively binds to a-mannopyranosyl and a-glucopyranosyl residues and might exhibit the brightest green fluorescence.

Platform


Fluorescence microscope

ExcitationFITC filter set
EmissionFITC 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™ 488-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™ 488-Concanavalin A Conjugate working solution (1X)
Add 5 µL of 200X 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™ 488-Concanavalin A working solution.
  3. Incubate cells with working solution for 10-30 minutes at 37 °C.
  4. Wash cells twice with HHBS buffer.
  5. Image cells on a fluorescence microscope using FITC 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™ 488-Concanavalin A working solution.
  3. Incubate cells with Concanavalin A conjugate working solution for 10-30 minutes at room temperature.
  4. Wash cells twice with HHBS buffer.
  5. Image cells on a fluorescence microscope using FITC filter set. 

Spectrum


Open in Advanced Spectrum Viewer
spectrum

Spectral properties

Correction Factor (260 nm)0.21
Correction Factor (280 nm)0.11
Extinction coefficient (cm -1 M -1)750001
Excitation (nm)491
Emission (nm)516
Quantum yield0.91

Product Family


NameExcitation (nm)Emission (nm)Extinction coefficient (cm -1 M -1)Quantum yieldCorrection Factor (260 nm)Correction Factor (280 nm)
iFluor® 555-Concanavalin A Conjugate55757010000010.6410.230.14
iFluor® 594-Concanavalin A Conjugate58760320000010.5310.050.04
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
Montelukast ameliorates Concanavalin A-induced autoimmune hepatitis in mice via inhibiting TNF-α/JNK signaling pathway.
Authors: El-Kashef, Dalia H and Abdelrahman, Rehab S
Journal: Toxicology and applied pharmacology (2020): 114931
Acupuncture stimulation attenuates TNF-α production via vagal modulation in the concanavalin A model of hepatitis.
Authors: Lim, Hee-Don and Kim, Ki-Joong and Jo, Byung Gon and Park, Ji-Yeun and Namgung, Uk
Journal: Acupuncture in medicine : journal of the British Medical Acupuncture Society (2020): 964528420907338
Deficiency of O-linked-glycosylation regulates activation of T cells and aggravates Concanavalin A-induced liver injury.
Authors: Hao, Xiaohua and Gao, Meixin and He, Lingling and Ye, Xiaohui and Yang, Junru and Zhang, Fuyang and Liu, Ran and Wei, Hongshan
Journal: Toxicology (2020): 152411
Nicotine attenuates concanavalin A-induced liver injury in mice by regulating the α7-nicotinic acetylcholine receptor in Kupffer cells.
Authors: Zhao, Jing and Park, Surim and Kim, Jong-Won and Qi, Jing and Zhou, Zixiong and Lim, Chae Woong and Kim, Bumseok
Journal: International immunopharmacology (2020): 106071
The high-resolution proteomic analysis of protein composition of rat spleen lymphocytes stimulated by Concanavalin A; a comparison with morphine-treated cells.
Authors: Ujcikova, Hana and Cechova, Kristina and Roubalova, Lenka and Brejchova, Jana and Kaufman, Jonas and Holan, Vladimir and Svoboda, Petr
Journal: Journal of neuroimmunology (2020): 577191
Surface-imprinted silica particles for Concanavalin A purification from Canavalia ensiformis.
Authors: Razym, Galiya and Bakhshpour, Monireh and Yavuz, Handan and Kip, Çiğdem and Tuncel, Ali and Denizli, Adil
Journal: Journal of chromatography. B, Analytical technologies in the biomedical and life sciences (2020): 121852
Pregabalin inhibits in vivo and in vitro cytokine secretion and attenuates spleen inflammation in Lipopolysaccharide/Concanavalin A -induced murine models of inflammation.
Authors: Abu-Rish, Eman Y and Mansour, Ahmad T and Mansour, Hebah T and Dahabiyeh, Lina A and Aleidi, Shereen M and Bustanji, Yasser
Journal: Scientific reports (2020): 4007
The Role of Serotonin in Concanavalin A-Induced Liver Injury in Mice.
Authors: Pang, Qing and Jin, Hao and Ke, Xiquan and Man, Zhongran and Wang, Yong and Tan, Yi and Lu, Zheng and Liu, Huichun
Journal: Oxidative medicine and cellular longevity (2020): 7504521
A Rabbit Model of Aqueous-Deficient Dry Eye Disease Induced by Concanavalin A Injection into the Lacrimal Glands: Application to Drug Efficacy Studies.
Authors: Honkanen, Robert A and Huang, Liqun and Rigas, Basil
Journal: Journal of visualized experiments : JoVE (2020)