Concanavalin a (cona)

Manufactured by Merck Group

Sourced in United States, Germany, United Kingdom, China, Macao, Japan, Spain, Sao Tome and Principe, Denmark, France, Switzerland, Italy, Canada, Australia, India

Concanavalin A is a lectin protein derived from the jack bean plant. It is commonly used as a research tool in biochemistry and cell biology laboratories. Concanavalin A has the ability to agglutinate (bind and aggregate) certain types of cells, making it useful for studying cell-surface carbohydrates and their interactions.

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Lab products found in correlation

1 331 protocols using concanavalin a (cona)

Investigating Galectin-9 in Concanavalin A-Induced Liver Injury

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Animal experiments were performed according to the guidelines of the Animal Care and Use Committee for Kagawa University (Takamatsu, Japan). Ethical approval was obtained from the Animal Care and Use Committee for Kagawa University. A total of 56 male BALB/c mice, at 7 weeks of age, 21–26 g, were purchased from Japan SLC, Inc. (Hamamatsu, Japan). The mice were housed together in a temperature-controlled environment under a 12 h:12 h day:night cycle (light from 07:00 to 19:00) at 22±1°C with freely available food and water throughout the experiment. Mice were given a single intravenous injection of ConA (Merck KGaA, Darmstadt, Germany) at a dose of 35 mg/kg body weight, and the animals were sacrificed 24 h following ConA administration or were observed for 48 h while checking for survival every 6 h. In the Gal-9 treated group (n=17), 00 µg Gal-9 (Department of Immunology, Faculty of Medicine, Kagawa University, Takamatsu, Japan) per mouse was injected subcutaneously immediately following the administration of ConA. Controls were treated with only PBS (n=10), only Gal-9 (n=10) or PBS with ConA (n=19).

Tadokoro T., Morishita A., Sakamoto T., Fujihara S., Fujita K., Mimura S., Oura K., Nomura T., Tani J., Yoneyama H., Iwama H., Himoto T., Niki T., Hirashima M, & Masaki T. (2017). Galectin-9 ameliorates fulminant liver injury. Molecular Medicine Reports, 16(1), 36-42.

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Yeast Cell Imaging on Concanavalin A

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Yeast were grown at 30°C in SD with 3% raffinose or SD with 2% glucose to an OD₆₀₀ of 0.4–0.6. For galactose induction, galactose was added to a final concentration of 3%. For imaging, 100-µl cell suspension was added to a 96-well glass-bottom plate (MGB096-1-2-LG-L; Brooks Life Science Systems) previously coated with concanavalin A (Con A) and concentrated on the bottom of the well by centrifugation. Wells were coated by adding 100 µl of 1 mg/ml Con A (Sigma-Aldrich) for 10 min before unbound Con A is removed and the Con A activated by adding 100 µl of 50-mM CaCl₂/50-mM MnSO₄ for 10 min. The solution was then removed, washed once with 100 µl ddH₂O, and air dried.

Saroufim M.A., Bensidoun P., Raymond P., Rahman S., Krause M.R., Oeffinger M, & Zenklusen D. (2015). The nuclear basket mediates perinuclear mRNA scanning in budding yeast. The Journal of Cell Biology, 211(6), 1131-1140.

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Yeast Cells Immobilization for SIM Imaging

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Yeast cells were grown at 30°C in SD with 2% glucose to an OD6₀₀ ~ 0.4–0.6. For imaging, 100-μl cell suspension was added to a 96-well glass-bottom plate (MGB096-1-2-LG-L; Brooks Life Science Systems) previously coated with concanavalin A (Con A) and concentrated on the bottom of the well by centrifugation. Wells were coated by adding 100 μl of 1 mg/ml Con A (Sigma-Aldrich) for 10 min before unbound Con A is removed and the Con A activated by adding 100 μl of 50 mM CaCl₂/50 mM MnSO₄ for 10 min. The solution was then removed, washed once with 100 μl ddH₂O, and air-dried. To minimize cell motion for SIM imaging, cells were briefly fixed with 70% EtOH for 5 min chilled at −20C, wash with cold 1x PBS before being added to glass-bottom plates coated with ConA.

Bensidoun P., Reiter T., Montpetit B., Zenklusen D, & Oeffinger M. (2022). Nuclear mRNA metabolism drives selective basket assembly on a subset of nuclear pore complexes in budding yeast. Molecular cell, 82(20), 3856-3871.e6.

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Concanavalin A-Induced Liver Injury Model

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Eight-week-old mice were intravenously (tail vein) injected with Concanavalin A (ConA) (Sigma, St. Louis MO), 15 mg/kg of body weight, diluted in sterile pyrogen-free PBS, as previously described ^{25 (link)}. 24h post injection, blood was collected by retro-orbital capillary plexus and hemolysis-free serum was generated by centrifugation of blood using serum separator tubes (Becton Dickinson, Franklin Lakes, NJ). Mice were then euthanized, liver weight was measured and collected for downstream analysis. Serum Alanine transaminase (ALT) and Aspartate Transaminase (AST) were analyzed at Missouri State University metabolic services core. A liver damage score from 1 to 4 was attributed macroscopically according to abnormal liver surface and to the extent of lesions. For chronic exposure to ConA, 8-week-old mice were intravenously (tail vein) injected with ConA (Sigma, 5 mg/kg of body weight, diluted in sterile pyrogen-free PBS) every week for 3 weeks, for a total of 3 injections. 48h post after the last injection, serum and tissues were collected and analyzed, as described above.

Etienne-Mesmin L., Vijay-Kumar M., Gewirtz A.T, & Chassaing B. (2016). Hepatocyte Toll-Like Receptor 5 Promotes Bacterial Clearance and Protects Mice Against High-Fat Diet–Induced Liver Disease. Cellular and Molecular Gastroenterology and Hepatology, 2(5), 584-604.

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Selection of Concanavalin A-Resistant Parasites

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PF 29-13 were grown in SDM-79 as described [13 (link),14 (link)]. To select for parasites that were spontaneously resistant to conA binding, lyophilized conA (Sigma, St. Louis, MO) was hydrated in cytomix [15 (link)] supplemented with 1 mM MnCl₂ (cytoM) and incubated at a final concentration of 2 μg/mL overnight with PF parasites. Agglutinated parasites were removed by slow speed centrifugation (40 x g, 10 min) and survivors allowed to recover 24 hr. This process was repeated three times, and then the concentration of conA increased to 5 μg/mL and then 10 μg/mL. Survivors were cloned by dilution and characterized.

Hackler A.L., Qiu Y., Patrick S.L., Hee Lee S., Acosta-Serrano A, & Morris J.C. (2015). Characterization of an African trypanosome mutant refractory to lectin-induced death. Biochemistry and Biophysics Reports, 4, 33-38.

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Lymphocyte Proliferation Assay using MTT

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We used a previously reported MTT assay procedure for lymphocyte proliferation assay (Babu et al., 2008[1 (link)]). Briefly, splenocytes were cultured in 96-well cell culture plates (Nunc, Denmark) and the aliquots of the antigens were then dispensed into the wells, at a final concentration of 10 μg/ml of antigen. Wells with splenocytes from mock-immunized mice were included as negative control. As well as ConA-stimulated cell suspensions were also included as positive control (ConA: Sigma-Aldrich, St. Louis, MO, USA). Cultures were incubated at 37 ºC in 5 % CO₂ humidified incubator for 1-3 days. The MTT proliferation assay was performed using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide. After incubation, cells were treated with MTT solvent for 15 minutes at RT. Absorbance was measured at OD = 570 nm.

Fahimi H., Sadeghizadeh M., Hassan Z.M., Auerswald H, & Schreiber M. (2018). Immunogenicity of a novel tetravalent dengue envelope protein domain III-based antigen in mice. EXCLI Journal, 17, 1054-1068.

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Isolation and Stimulation of Rat PBMCs

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Peripheral blood mononuclear cells were isolated from whole blood samples of the experimental rats by Percoll (Sigma-Aldrich, St Louis, MO, USA) gradient centrifugation at 400g for 25 min. The PBMCs were seeded in 96-well plates at 10⁶ cells/well and were cultured in 1640 RPMI medium containing 10% fetal bovine serum (FBS, Pan, Germany), 100 U/ml penicillin and 100 μg/ml streptomycin (Beyotime Biotechnology, China) at 37 °C. The cells were divided into five groups. In the stimulation group, the cells were stimulated with 100 μg/ml Concanavalin A (Con A, Sigma-Aldrich, St. Louis, MO, USA) for 24 h. In the treatment groups, the cells were incubated with different concentrations of ImKTx88 (100 nM, 1, and 10 μM) 60 min before Con A stimulation. The control cells were given PBS treatment without any stimulation. Those cells were cultured for 24 h before harvesting.

Huang J., Han S., Sun Q., Zhao Y., Liu J., Yuan X., Mao W., Peng B., Liu W., Yin J, & He X. (2017). Kv1.3 channel blocker (ImKTx88) maintains blood–brain barrier in experimental autoimmune encephalomyelitis. Cell & Bioscience, 7, 31.

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Stimulation of PBMCs for RNA Extraction

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PBMCs were thawed, washed twice, and incubated in complete medium (RPMI 1640 with 10% FBS) at 37 °C and 5% CO₂. The cells were then washed, counted, suspended in complete medium, and seeded at 1 × 10⁶ cells/well on 24 well plates and incubated 1 h at 37 °C and 5% CO₂ prior to stimulation. Stimulation was performed with Concanavalin A (Sigma-Aldrich) (ConA) (2.5 µg/ml), Map sonicate (SM) (10 µg/ml) or left untreated (PBS). Cultures were incubated for 24 h at 37 °C °C and 5% CO₂ and centrifuged at 300 × g for 8 min for RNA extraction.

Ladero-Auñon I., Molina E., Oyanguren M., Barriales D., Fuertes M., Sevilla I.A., Luo L., Arrazuria R., De Buck J., Anguita J, & Elguezabal N. (2021). Oral vaccination stimulates neutrophil functionality and exerts protection in a Mycobacterium avium subsp. paratuberculosis infection model. NPJ Vaccines, 6, 102.

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Spleen Cell Proliferation Assay for Immunology

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Briefly, the spleens of five mice from each group were aseptically removed 2 weeks after the last immunization. Spleen cells were harvested by pushing the spleen through a nylon sieve and then red blood cells were removed using red blood cell lysis buffer (Sigma, St. Louis, MO, USA). The purified spleen cells were resuspended in Dulbecco's modified Eagle's medium supplemented with 10% fetal bovine serum. Proliferation of spleen lymphocytes was detected using a CCK-8 kit (Dojindo, Kumamoto, Japan) according to the manufacturer's instructions. Purified spleen lymphocytes were added to 96-well-plates and when their density reached 2 ^* 10⁵, lymphocytes were stimulated with rGRA24 (10 μg/ml), Concanavalin A as a positive control (ConA; 5 μg/ml; Sigma) or Dulbecco's modified Eagle's medium alone as a negative control. Four days later, CCK-8 (Dojindo) was added to each well and incubated for 4 h to stimulate lymphocyte proliferation. The stimulation index (SI) was calculated using the following formula: stimulation index (SI) = (OD₅₇₀ rGRA24/OD₅₇₀ Control):(OD₅₇₀ ConA/OD₅₇₀ Control).

Zheng B., Lou D., Ding J., Zhuo X., Ding H., Kong Q, & Lu S. (2019). GRA24-Based DNA Vaccine Prolongs Survival in Mice Challenged With a Virulent Toxoplasma gondii Strain. Frontiers in Immunology, 10, 418.

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Gasdermin D Knockout Mice in Autoimmune Hepatitis

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Gsdmd^−/− mice were purchased from the Model Animal Research Center of Nanjing University and bred in our facility. Mice aged 6 to 8 weeks were used in this experiment. Littermate WT mice were used as a control for genetic knockout mice. Mice were divided into four groups: WT+NS, WT+ConA, Gsdmd^−/−+NS, and Gsdmd^−/−+ConA. Mice in the ConA group were challenged with 15 mg/kg ConA (Sigma-Aldrich) through tail vein injection to establish AIH. Mice in the NS group received an equal amount of 0.9% normal saline (NS). Mice were sacrificed 8 h after treatment, and feces, blood, liver, and colon tissues were collected. All experiments were approved by the Animal Care and Use Committee of the First Affiliated Hospital, School of Medicine, Zhejiang University.

Wang K., Wu W., Jiang X., Xia J., Lv L., Li S., Zhuge A., Wu Z., Wang Q., Wang S, & Li L. (2022). Multi-Omics Analysis Reveals the Protection of Gasdermin D in Concanavalin A-Induced Autoimmune Hepatitis. Microbiology Spectrum, 10(5), e01717-22.

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