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iFluor® 488-Concanavalin A Conjugate
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.
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.
Note The reconstituted conjugate solution can be stored at 2-8 °C for short-term storage or at -20 °C for long-term storage.
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
- Wash cells twice with a HHBS buffer.
- Add 100 µL iFluor™ 488-Concanavalin A working solution.
- Incubate cells with working solution for 10-30 minutes at 37 °C.
- Wash cells twice with HHBS buffer.
- Image cells on a fluorescence microscope using FITC filter set.
Fixed Cells Stain
Concanavalin A conjugates can be also used to stain fixed cells.- 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. - Add 100 µL iFluor™ 488-Concanavalin A working solution.
- Incubate cells with Concanavalin A conjugate working solution for 10-30 minutes at room temperature.
- Wash cells twice with HHBS buffer.
- Image cells on a fluorescence microscope using FITC filter set.
Spectrum
Product family
| Name | Excitation (nm) | Emission (nm) | Extinction coefficient (cm -1 M -1) | Quantum yield | Correction Factor (260 nm) | Correction Factor (280 nm) |
| iFluor® 555-Concanavalin A Conjugate | 557 | 570 | 1000001 | 0.641 | 0.23 | 0.14 |
| iFluor® 594-Concanavalin A Conjugate | 587 | 603 | 2000001 | 0.531 | 0.05 | 0.04 |
| iFluor® 647-Concanavalin A Conjugate | 656 | 670 | 2500001 | 0.251 | 0.03 | 0.03 |
| iFluor® 750-Concanavalin A Conjugate | 757 | 779 | 2750001 | 0.121 | 0.044 | 0.039 |
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
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
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
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
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
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
Page updated on October 22, 2024
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