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> Chemicals & Drugs > Amino Acid > Cholera Toxin

Cholera Toxin

Cholera Toxin is a potent enterotoxin produced by the bacterium Vibrio cholerae, the causative agent of the severe diarrheal disease cholera.
This toxin is a key virulence factor that disrupts fluid and electrolyte homeostasis in the intestinal epithelium, leading to the profuse watery diarrhea characteristic of cholera.
Cholera Toxin consists of two main subunits: the enzymatically active A subunit and the binding B subunit.
The B subunit mediates cellular uptake, while the A subunit catalyzes the ADP-ribosylation of the Gs alpha subunit of adenylate cyclase, resulting in constitutive activation and increased cAMP production.
This cascade ultimately leads to the dysregulation of ion channels and pumps, causing the massive fluid efflux seen in cholera.
Cholera Toxin is an important tool in biomedical research, used to study signal transduction pathways, as well as a potent adjuvant in vaccine development due to its ability to enhance immune responses.
Understanding the molecular mechanisms of Cholera Toxin is crucial for improving prevention and treatment strategies agaisnt this devastating disease.

Most cited protocols related to «Cholera Toxin»

1
Fractionation of Breast Epithelial Cell Proteome
Cited 65 times
Human breast epithelial cells (MCF10A) were transfected with a pQCXIH vector and were cultured in DMEM/F12 (Invitrogen, Carlsbad, CA) supplemented with 5% horse serum (Invitrogen), 20 ng/mL EGF, 0.5 μg/mL hydro-cortisone, 100 ng/mL cholera toxin, 10 μg/mL insulin, and 50 μg/mL penicillin/streptomycin until 70–80% confluence was reached. The cells were then lysed in a buffer containing 8 M urea, 2.5 mM sodium pyrophosphate, 1 mM β-glycerophosphate, 50 mM ammonium bicarbonate, one-third tablet of protease inhibitor, and 2 mM sodium ortho-vanadate. Proteins were denatured, reduced, and alkylated after which tryptic digestions were performed at an enzyme/substrate ratio of 1:50. For method comparison between SCX and RP, digested peptides were cleaned by flowing through a 1 mL solid-phase extraction C18 column (Discovery DSC-18, SUPELCO, Bellefonte, PA). Samples were concentrated using a Speed-Vac SC 250 Express (Thermo Savant, Holbrook, NY) and stored at −80°C until time for analysis. A 300.0 μg desalted peptide sample was used for each SCX, low-pH RPLC, and high-pH RPLC fractionation. A 300.0 μg nondesalted protein digest was used to evaluate the potential of high-pH approach for desalting.
Wang Y., Yang F., Gritsenko M.A., Wang Y., Clauss T., Liu T., Shen Y., Monroe M.E., Lopez-Ferrer D., Reno T., Moore R.J., Klemke R.L., Camp DG I.I, & Smith R.D. (2011). Reversed-phase chromatography with multiple fraction concatenation strategy for proteome profiling of human MCF10A cells. Proteomics, 11(10), 2019-2026.
Publication 2011
ammonium bicarbonate beta-glycerol phosphate Breast Buffers Cells Cholera Toxin Cloning Vectors Cortisone Digestion Enzymes Epithelial Cells Equus caballus Fractionation, Chemical Homo sapiens Insulin Penicillins Peptides Protease Inhibitors Proteins Serum sodium pyrophosphate Sodium Vanadate Solid Phase Extraction Streptomycin Tablet Trypsin Urea
2
Mammary Epithelial Cell Isolation and Culture
Cited 64 times
Finite lifespan pre-stasis HMEC from specimens 184 (batch D), 48R (batch T), and 240L (batch B), and post-selection HMEC 184 (batch B, agonescence at ~passage 15), 48R (batch S, agonescence at ~passage 23), and 240L (agonescence at ~passage 18) were obtained from reduction mammoplasty tissue of women aged 21,16, and 19 respectively. Cells were initiated as organoids in primary culture in either serum-free MCDB170 medium (MEGM, Lonza, Walkersville, MD) plus supplements (2 (link)), or serum-containing media MM (1 (link)) or M85, and subjected to multiple partial trypsinizations, as described (18 ). Post-selection HMEC were cultured in MCDB170 as described (2 (link),18 ,19 ). M85 medium is composed of 50% MM medium and 50% bicarbonate-free supplemented MCDB170; M87 medium is composed of 50% MM4 (MM without the conditioned media)(1 (link)) and 50% supplemented MCDB170. Cholera toxin was added to M85 and M87 at a final concentration of 0.5 ng/ml and oxytocin (Bachem) at 0.1 nM. M85A and M87A media were supplemented with 0.1% AlbuMAX I (Invitrogen). Fibroblasts from specimens 184, 48, and 240L were obtained by growing primary reduction mammoplasty cells in DME/F12 with 10% FBS and 10 μg/ml insulin. 250MK cells were obtained from aspirated milk fluids1.
Total PD level for each culture was calculated beginning at passage 2 using the formula PD=log2(Nfinal/Ninitial) where Ninitial is the number of cells seeded in a dish at each passage and Nfinal is the number of cells recovered from the dish. No corrections were made for plating efficiency.
Garbe J.C., Bhattacharya S., Merchant B., Bassett E., Swisshelm K., Feiler H.S., Wyrobek A.J, & Stampfer M.R. (2009). Molecular distinctions between the stasis and telomere attrition senescence barriers demonstrated by long-term culture of normal human mammary epithelial cells. Cancer research, 69(19), 7557-7568.
Publication 2009
Cells Cholera Toxin Culture Media, Conditioned Fibroblasts Hyperostosis, Diffuse Idiopathic Skeletal Insulin Ion, Bicarbonate Mammaplasty Milk, Cow's Organoids Oxytocin Serum Tissues Woman
3
Establishment of Primary Breast Cancer Organoids
Cited 21 times

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Publication 2013
Antibiotics Centrifugation Cholera Toxin Collagenase Collagen Type I Common Cold Deoxyribonucleases Hydrocortisone Insulin Malignant Neoplasm of Breast Mammary Epithelia, Human matrigel Neoplasms Organoids Trypsin
4
Mouse Models of Helicobacter Pylori Infection
Cited 16 times

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Publication 2010
Antibiotics Antibodies, Anti-Idiotypic Biological Factors bismuth subcitrate Body Weight Cells Cholera Toxin Diphtheria Toxin Helicobacter pylori IL2RA protein, human IL10 protein, human Infection Metronidazole Microscopy Mus Regulatory T-Lymphocytes Rivers Strains Tetracycline Hydrochloride Translocation, Chromosomal Vaccination
5
Establishment and Characterization of Ovarian Cancer Cell Lines
Cited 14 times
The OCI cell lines and OCMI medium are available from the Live Tumor Culture Core (LTCC) at Sylvester Comprehensive Cancer Center (SCCC), Miller School of Medicine, University of Miami http://sylvester.org/shared-resources/Live-Tumor-Culture-Core In order to establish the cell lines, fresh tumor tissue fragments are minced and plated on Primaria (BD Biosciences) plates before and after digestion with 1 mg/ml collagenase (Roche). The tumor cells are cultured in nutrient medium described previously 16 (link), supplemented with insulin, hydrocortisone, EGF, cholera toxin, and serum as described in Supplementary Methods. We refer to this version of the medium as Ovarian Carcinoma Modified Ince medium (OCMI). This formulation is supplemented with 17β-estradiol for endometrioid and mucinous tumors (OCMIe). The papillary serous, clear cell, dysgerminoma, and carcinosarcoma tumors are initially cultured in 5% CO2 and regular O2 at 37 C as monolayers attached to Primaria culture plates. The endometrioid and mucinous tumors are initially cultured in 5% CO2 and low O2 (5%) at 37 C as monolayers attached to Primaria culture plates. Adjustments in O2 and plates are made for individual cell lines as necessary (Supplementary Table 3). The tumor cells are passaged at a ~ 1:3 ratio once a week and plated into a new flask at approximately 1×104 cells/cm2. During the initial weeks of culture, (~1–5) the plates are treated with diluted trypsin first, in order to deplete stromal cells. The remaining cells that are still attached to the culture plate are treated with 0.25% trypsin for sub-culturing. In general, tumor cultures are free of stromal and normal cell types within 4–6 passages. The SOC cell lines were STR validated and cultured as per the instructions of the vendor. OCI lines will be available from the Ince laboratory upon publication. All study procedures were approved by the Institutional Review Boards of the University of Miami, Brigham & Women’s and Massachusetts General Hospitals to collect discarded tissues with written consent from all patients. See Supplementary Note 1 for further details of culture methods.
Ince T.A., Sousa A.D., Jones M.A., Harrell J.C., Agoston E.S., Krohn M., Selfors L.M., Liu W., Chen K., Yong M., Buchwald P., Wang B., Hale K.S., Cohick E., Sergent P., Witt A., Kozhekbaeva Z., Gao S., Agoston A.T., Merritt M.A., Foster R., Rueda B.R., Crum C.P., Brugge J.S, & Mills G.B. (2015). Characterization of twenty-five ovarian tumour cell lines that phenocopy primary tumours. Nature Communications, 6, 7419.
Publication 2015
Carcinosarcoma Cell Lines Cells Cholera Toxin Collagenase Culture Techniques Digestion Dysgerminoma Endometrium Estradiol Ethics Committees, Research Hydrocortisone Insulin Malignant Neoplasms Neoplasms Neoplasms, Mucinous Nutrients Ovarian Cancer Patients Serum Stromal Cells Tissues Trypsin Woman

Most recents protocols related to «Cholera Toxin»

1
Immortalized Breast Cell Culture
Human mammary immortalized cells (MCF10A), and the TNBC cell line MDA-MB-231 were purchased from American Type Culture Collection. MCF10A cells were grown in a 1:1 mixture of Dulbecco’s modified Eagle’s medium (DMEM) and Ham’s F12 medium with 20 ng mL−1 human epidermal growth factor, 100 ng mL−1 cholera toxin, 0.01 mg mL−1 bovine insulin, 500 ng mL−1 hydrocortisone, and 5% horse serum. MDA-MB-231 cells were grown in DMEM containing L-glutamine and sodium pyruvate, supplemented with 10% fetal bovine serum and 1% antibiotic and antimycotic solution in a humidified atmosphere of 5% CO2 at 37°C in an incubator. Human BCSC cells: ALDH+ and CD44+/CD24−, and BC cells: ALDH- were purchased from Celprogen (San Pedro, CA, United States) and maintained in human BCSC expansion and undifferentiation media. DAPT (γ-secretase), cycloheximide (CHX), chloroquine (CQ), and MG132 were purchased from Sigma (St. Louis, MO).
Saran U., Chandrasekaran B., Tyagi A., Shukla V., Singh A., Sharma A.K, & Damodaran C. (2023). A small molecule inhibitor of Notch1 modulates stemness and suppresses breast cancer cell growth. Frontiers in Pharmacology, 14, 1150774.
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Publication 2023
1,2-dilinolenoyl-3-(4-aminobutyryl)propane-1,2,3-triol Antibiotics Atmosphere Bos taurus Breast CD44 protein, human Cell Culture Techniques Cell Lines Cells Chloroquine Cholera Toxin Cycloheximide Eagle Epidermal growth factor Equus caballus Fetal Bovine Serum Glutamine Homo sapiens Hydrocortisone Insulin MDA-MB-231 Cells MG 132 Pyruvate Secretase Serum Sodium
2
Culturing Human Cell Lines for Research
Normal human mammary epithelial cells MCF-10A (cat. no. MZ-0695), and breast cancer cells MCF-7 (cat. no. MZ-0113), BT-549 (cat. no. MZ-0031) and MDA-MB-231 (cat. no. 228845) were obtained from Ningbo Mingzhou Biotechnology Co., Ltd. Human umbilical vein endothelial cells (HUVECs; cat. no. CL-0122) were provided by PromoCell GmbH and passaged in endothelial cell growth medium (PromoCell GmbH). MCF-10A cells were cultured in mammary epithelium growth medium (Shanghai Yaji Biological Technology Co., Ltd.) with 100 ng/ml cholera toxin and 1% Penicillin/Streptomycin. BT-549 cells were cultured in RPMI-1640 medium (Gibco; Thermo Fisher Scientific, Inc.) with 0.023 U/ml insulin and 10% fetal bovine serum (FBS; Gibco; Thermo Fisher Scientific, Inc.). MCF-7 and MDA-MB-231 cells were cultured in DMEM with 10% FBS, 50 IU/ml penicillin and 50 µg/ml streptomycin. Human monocytic leukemia THP-1 cells were purchased from the Type Culture Collection of the Chinese Academy of Sciences and cultured in RPMI-1640 medium supplemented with 10% FBS (Gibco; Thermo Fisher Scientific, Inc.). All cells were maintained at 37˚C in humidified air with 5% CO2. Two culture wells were employed for each experiment, including one for the experimental group and the other one for the control group.
Zheng Y., Wang S., Zhong Y., Huang C, & Wu X. (2023). A20 affects macrophage polarization through the NLRP3 inflammasome signaling pathway and promotes breast cancer progression. Experimental and Therapeutic Medicine, 25(4), 147.
Publication 2023
Biopharmaceuticals Breast Breast Carcinoma Cell Culture Techniques Cells Chinese Cholera Toxin Culture Media Endothelial Cells Epithelial Cells Epithelium Homo sapiens Human Umbilical Vein Endothelial Cells Insulin Leukemia MDA-MB-231 Cells Monocytes Penicillins Streptomycin THP-1 Cells
3
Immortalization of Primary Cells

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Publication 2023
Adenine Cell Culture Techniques Cells Cholera Toxin Eagle Embryo Feeder Cell Layers Fibroblasts Hydrocortisone Insulin Mus Nutrients Y 27632
4
Preparation and Preservation of Cultured Epidermis

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Publication 2023
Aftercare Antibiotics, Antitubercular Atmosphere Bath Cholera Toxin Cryoprotective Agents dispase II Epidermal growth factor Feeder Layer Cells Fetal Bovine Serum Homo sapiens Humidity Hydrocortisone Insulin Keratinocyte Liothyronine Normal Saline Physical Processes Skin Sulfoxide, Dimethyl
5
Culturing Metastatic and Non-Tumorigenic Breast Cells
Metastatic MDA-MB-231 and non-tumorigenic MCF-10A human breast cell lines were purchased from American Type Cell Culture Collection (ATCC) and cultured using standardized media and conditions as previously described31 (link). Briefly, MDA-MB-231 cells were cultured in DMEM (Corning, No. 10013CV), supplemented with 10% fetal bovine serum (ATCC, No. 302020). MCF-10A cells were cultured in DMEM/F-12 (ThermoFisher, No. 11330032) supplemented with 5% horse serum (Invitrogen, No. 16050122), 20 ng/ml EGF (Peprotech, AF10015; ThermoFisher, No. 10605HNAE), 0.5 mg/ml hydrocortisone (Sigma-Aldrich, No. H0888), 100 ng/ml choleratoxin (Sigma-Aldrich, C8052), 10 mg/ml insulin (Sigma-Aldrich, No. I1882). Both media recipes contained 1% penicillin/streptomycin (ATCC, No. 302300; ThermoFisher, No. 15140122). Cells were maintained at 37 °C and 5% CO2 in a cell culture incubator.
Karrobi K., Tank A., Fuzail M.A., Kalidoss M., Tilbury K., Zaman M., Ferruzzi J, & Roblyer D. (2023). Fluorescence Lifetime Imaging Microscopy (FLIM) reveals spatial-metabolic changes in 3D breast cancer spheroids. Scientific Reports, 13, 3624.
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Publication 2023
Breast Cell Culture Techniques Cell Lines Cells Cholera Toxin Equus caballus Fetal Bovine Serum Homo sapiens Hydrocortisone Insulin MDA-MB-231 Cells Neoplastic Cell Transformation Penicillins Serum Streptomycin

Top products related to «Cholera Toxin»

1
Cholera toxin by Merck Group
Sourced in United States, Germany, United Kingdom, Italy, France, China, Canada, Macao, Sao Tome and Principe, Australia, Israel, Switzerland, Spain, Japan
Cholera toxin is a bacterial protein produced by the bacterium Vibrio cholerae. It has a well-documented function as a potent activator of the adenylate cyclase enzyme, leading to increased levels of cyclic AMP (cAMP) in target cells. This property makes cholera toxin a valuable tool in various areas of biological research, such as cell signaling studies and the investigation of cellular regulatory mechanisms.
2
Hydrocortisone by Merck Group
Sourced in United States, Germany, United Kingdom, France, China, Italy, Canada, Macao, Japan, Israel, Switzerland, Australia, Sao Tome and Principe, Spain, Austria, Portugal, Belgium, Denmark, Sweden, Argentina, Brazil, Poland, New Zealand
Hydrocortisone is a laboratory-grade reagent used in various research and analytical applications. It is a synthetic corticosteroid compound with anti-inflammatory and immunosuppressant properties. Hydrocortisone is commonly utilized as a standard or reference material in analytical procedures, such as assays and chromatographic techniques, to quantify and identify related compounds.
3
Insulin by Merck Group
Sourced in United States, Germany, United Kingdom, China, France, Canada, Italy, Sao Tome and Principe, Japan, Switzerland, Macao, Israel, Australia, Spain, Austria, Sweden, Poland, Denmark, New Zealand, Belgium, Portugal, Ireland, Netherlands, Brazil, Colombia, India, Morocco, Argentina
Insulin is a lab equipment product designed to measure and analyze insulin levels. It provides accurate and reliable results for research and diagnostic purposes.
4
Horse serum by Thermo Fisher Scientific
Sourced in United States, United Kingdom, Germany, New Zealand, Japan, China, France, Australia, Italy, Spain, Switzerland, Canada, Netherlands, Denmark, Austria, Belgium, Ireland, Israel, Brazil
Horse serum is a biological fluid derived from the blood of horses. It contains a complex mixture of proteins, including immunoglobulins, hormones, and other biomolecules. Horse serum is commonly used as a supplement in cell culture media to support the growth and maintenance of various cell types.
5
Fetal bovine serum (fbs) by Thermo Fisher Scientific
Sourced in United States, China, United Kingdom, Germany, Australia, Japan, Canada, Italy, France, Switzerland, New Zealand, Brazil, Belgium, India, Spain, Israel, Austria, Poland, Ireland, Sweden, Macao, Netherlands, Denmark, Cameroon, Singapore, Portugal, Argentina, Holy See (Vatican City State), Morocco, Uruguay, Mexico, Thailand, Sao Tome and Principe, Hungary, Panama, Hong Kong, Norway, United Arab Emirates, Czechia, Russian Federation, Chile, Moldova, Republic of, Gabon, Palestine, State of, Saudi Arabia, Senegal
Fetal Bovine Serum (FBS) is a cell culture supplement derived from the blood of bovine fetuses. FBS provides a source of proteins, growth factors, and other components that support the growth and maintenance of various cell types in in vitro cell culture applications.
6
Dmem f12 by Thermo Fisher Scientific
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DMEM/F12 is a cell culture medium developed by Thermo Fisher Scientific. It is a balanced salt solution that provides nutrients and growth factors essential for the cultivation of a variety of cell types, including adherent and suspension cells. The medium is formulated to support the proliferation and maintenance of cells in vitro.
7
Epidermal growth factor (egf) by Thermo Fisher Scientific
Sourced in United States, United Kingdom, Germany, China, Canada, France, Italy, Japan, Israel, Switzerland, Australia, Macao, Belgium, Spain, Denmark, Jersey
EGF is a lab equipment product from Thermo Fisher Scientific. It is a recombinant human Epidermal Growth Factor (EGF) protein. EGF is a growth factor that plays a role in cell proliferation and differentiation.
8
Penicillin streptomycin by Thermo Fisher Scientific
Sourced in United States, Germany, United Kingdom, China, Canada, France, Japan, Australia, Switzerland, Israel, Italy, Belgium, Austria, Spain, Gabon, Ireland, New Zealand, Sweden, Netherlands, Denmark, Brazil, Macao, India, Singapore, Poland, Argentina, Cameroon, Uruguay, Morocco, Panama, Colombia, Holy See (Vatican City State), Hungary, Norway, Portugal, Mexico, Thailand, Palestine, State of, Finland, Moldova, Republic of, Jamaica, Czechia
Penicillin/streptomycin is a commonly used antibiotic solution for cell culture applications. It contains a combination of penicillin and streptomycin, which are broad-spectrum antibiotics that inhibit the growth of both Gram-positive and Gram-negative bacteria.
9
Mcf 10a by American Type Culture Collection
Sourced in United States, China, United Kingdom, Germany, France, Canada, Switzerland
MCF-10A is a non-tumorigenic human breast epithelial cell line. It is a well-characterized model for studying normal breast cell biology.
10
Epidermal growth factor (egf) by Merck Group
Sourced in United States, Germany, United Kingdom, China, Italy, Canada, Japan, Spain, France, Macao, Israel, Australia, Denmark, Sao Tome and Principe, Morocco, Austria, Portugal, Belgium, Poland, Switzerland
EGF is a laboratory equipment product manufactured by Merck Group. It is a protein that plays a key role in cell growth and differentiation processes. EGF primarily functions as a growth factor, stimulating cell division and proliferation in various cell types.

More about "Cholera Toxin"

Cholera Toxin (CT) is a potent enterotoxin produced by the bacterium Vibrio cholerae, the causative agent of the severe diarrheal disease cholera.
This key virulence factor is a critical component in the pathogenesis of cholera, disrupting fluid and electrolyte homeostasis in the intestinal epithelium and leading to the characteristic profuse watery diarrhea.
CT consists of two main subunits: the enzymatically active A subunit and the binding B subunit.
The B subunit mediates cellular uptake, while the A subunit catalyzes the ADP-ribosylation of the Gs alpha subunit of adenylate cyclase, resulting in constitutive activation and increased cAMP production.
This cascade ultimately leads to the dysregulation of ion channels and pumps, causing the massive fluid efflux seen in cholera.
Beyond its role in cholera, CT is an important tool in biomedical research, used to study signal transduction pathways and as a potent adjuvant in vaccine development due to its ability to enhance immune responses.
Understanding the molecular mechanisms of CT is crucial for improving prevention and treatment strategies against this devastating disease.
In related research, Hydrocortisone, Insulin, Horse serum, FBS, DMEM/F12, EGF, and Penicillin/streptomycin are commonly used in cell culture experiments involving the MCF-10A cell line, which is a non-tumorigenic human breast epithelial cell line.
These components are essential for maintaining the growth and differentiation of this cell line, which is often used as a model for studying breast cancer and related pathways.