Ca2 ionomycin

Manufactured by Merck Group

Sourced in United States

Ca2+ ionomycin is a chemical compound used as a laboratory tool for research purposes. It functions as a calcium ionophore, facilitating the movement of calcium ions across cell membranes.

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Lab products found in correlation

Brefeldin a, by Bio-Rad (2 mentions) Anti cd8 pacific blue, by Bio-Rad (2 mentions) Anti cd4 fitc, by Bio-Rad (2 mentions) Cytofix cytoperm kit, by BD (2 mentions) Golgiplug, by BD (2 mentions) Cytochalasin b, by Merck Group (1 mentions) Canine ifn γ elispot kit, by R&D Systems (1 mentions) Anti cd3, by Bio-Rad (1 mentions)

Close spelling variants of this product

Ionomycin (3612 citations)

4 protocols using ca2 ionomycin

Quantifying IFN-γ Production in T Cells

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For assaying IFNγ production, 4 × 10⁵ PBMCs were stimulated for 5 h in the presence of 2 ng/mL PMA, 4 μg/mL Ca²⁺ ionomycin (Sigma-Aldrich), and Brefeldin A (BD GolgiPlug; BD Biosciences, San Jose, CA, USA) adapted from described stimulation conditions [32 (link)] or overnight with 40 μg/mL T. cruzi lysate at 37 °C. Brefeldin A was added 5 h prior to end of incubation, and the cells were stained with anti-CD8-Pacific Blue and anti-CD4-FITC (AbD Serotec, Raleigh, NC, USA) followed by intracellular staining with anti-bovine IFN-γ AF647 (AbD Serotec) according to the BD Cytofix/Cytoperm kit (BD Biosciences). Samples were fixed in 2% formaldehyde prior to analysis by flow cytometry.

Hartley A.N., Cooley G., Gwyn S., Orozco M.M, & Tarleton R.L. (2014). Frequency of IFNγ-producing T cells correlates with seroreactivity and activated T cells during canine Trypanosoma cruzi infection. Veterinary Research, 45(1), 6.

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Porcine Parthenogenetic Embryo Culture

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Following maturation, cumulus cells were removed by pipetting in the presence of 1 mg/mL hyaluronidase for 2 to 3 min. Oocytes were parthenogenetically activated with 5 μM Ca²⁺ ionomycin (Sigma) for 5 min. After 3 h of culture in porcine zygote medium-5 (PZM-5) containing 7.5 μg/mL cytochalasin B (Sigma), embryos were washed three times in PZM-5 containing 0.4% (w/v) BSA and cultured in the same medium for 7 days at 38.8°C in a humidified atmosphere of 5% CO₂ and 95% air. The oocytes and embryos were washed in DPBS, and depending on the experiment, either fixed in 3.7% (w/v) paraformaldehyde for 20 min and stored at 4°C, or snap-frozen in liquid nitrogen and stored at −70°C.

Lee S.E., Kim E.Y., Choi H.Y., Moon J.J., Park M.J., Lee J.B., Jeong C.J, & Park S.P. (2014). Rapamycin Rescues the Poor Developmental Capacity of Aged Porcine Oocytes. Asian-Australasian Journal of Animal Sciences, 27(5), 635-647.

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T. cruzi-Induced IFNγ ELISpot Assay

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Four hundred thousand PBMCs were cultured in media, 2 ng/mL phorbol 12-myristate 13-acetate (PMA) and 500 ng/mL Ca²⁺ ionomycin (both Sigma-Aldrich, St. Louis, MO, USA), or 10 μg/mL T. cruzi amastigote lysate for 16 h at 37 °C and 5% CO₂. Cells were assayed for IFNγ production using the Canine IFN-γ ELISpot kit (cat no. EL781, R& D Systems, Inc, Minneapolis, MN, USA) following standard kit protocol. Enumeration of spot forming cells was completed by an ImmunoSpot analyzer (CTL, Cleveland, OH, USA). Mean numbers of spots from duplicate or triplicate well were obtained for media and T. cruzi lysate stimulation conditions. Responses were considered positive if a minimum of 20 spots/10⁶ PBMC total were present and this number was at least twice the value of wells assayed with media alone [27 (link)].

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Canine Vaccine IFNγ Response Assay

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For assaying IFNγ levels, 4×10⁵ PBMCs were stimulated for 5hr in the presence of 2ng/mL phorbol 12-myristate 13-acetate (PMA), 4µg/mL Ca²⁺ ionomycin (Sigma-Aldrich), and brefeldin A (BD GolgiPlug; BD Biosciences) (Pedersen et al., 2002 ). For polyclonal activation, PBMCs were plated with 15µg/mL anti-CD3 (AbD Serotec) or diluted whole vaccine antigens, incubated overnight at 37°C and brefeldin A added 5hr prior to end of incubation. Canine vaccines were IMRAB 3TF (Rabies; Merial, Athens, GA, USA), Duramune 5 (Canine distemper-Adenovirus Type 2-Parainfluenza-Parvovirus (DAPP); Fort Dodge Animal Health, Fort Dodge, IA, USA), and Leptovax 4 (Leptospirosis bacterial extract; Fort Dodge Animal Health) vaccines. Cells were stained with anti-CD8-Pacific Blue and anti-CD4-FITC (AbD Serotec) followed by intracellular staining with anti-bovine anti-IFN-γ AF647 (AbD Serotec) according to the BD Cytofix/Cytoperm kit (BD Biosciences).

Hartley A.N, & Tarleton R.L. (2015). CHEMOKINE RECEPTOR 7 (CCR7)-EXPRESSION AND IFNγ PRODUCTION DEFINE VACCINE-SPECIFIC CANINE T CELL SUBSETS. Veterinary immunology and immunopathology, 164(0), 127-136.

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