MW 747.1 Da, Purity >99%. Ca2+ ionophore. Useful when calcium-dose response data are not required. Ion specificity Mn2+>Ca2+>Mg2+>Sr2+>Ba2+.
MW 747.1 Da, Purity >99%. Ca2+ ionophore. Useful when calcium-dose response data are not required. Ion specificity Mn2+>Ca2+>Mg2+>Sr2+>Ba2+.
Ca2+ ionophore. Useful when calcium-dose response data are not required. Ion specificity Mn2+>Ca2+>Mg2+>Sr2+>Ba2+.
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2D chemical structure image of ab120116, Ionomycin Ca2+ Salt, Ca2+ ionophore
Histone H3 (citrulline R8) was immunoprecipitated from 0.35 mg of HEK-293T (Human epithelial cell line from embryonic kidney) transfected with PADI4 (WT), then treated with 10mM CaCl2 and 10μM Ionomycin (ab120116) for 2 hours, whole cell lysate with Anti-Histone H3 (citrulline R8) antibody [EPR20358-13] ab219406 at 1/30 dilution.
Western blot was performed from the immunoprecipitate using Anti-Histone H3 (citrulline R8) antibody [EPR20358-13] ab219406 at 1/1000 dilution.
VeriBlot for IP Detection Reagent (HRP) (VeriBlot for IP Detection Reagent (HRP) ab131366), was used for detection at 1/10000 dilution.
Lane 1: HEK-293T transfected with PADI4 (WT), then treated with 10mM CaCl2 and 10μM Ionomycin (ab120116) for 2 hours, whole cell lysate 10 μg (Input).
Lane 2: Anti-Histone H3 (citrulline R8) antibody [EPR20358-13] ab219406 IP in HEK-293T transfected with PADI4 (WT), then treated with 10mM CaCl2 and 10μM Ionomycin (ab120116) for 2 hours, whole cell lysate.
Lane 3: Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) instead of Anti-Histone H3 (citrulline R8) antibody [EPR20358-13] ab219406 in HEK-293T transfected with PADI4 (WT), then treated with 10mM CaCl2 and 10μM Ionomycin (ab120116) for 2 hours, whole cell lysate.
Blocking and dilution buffer: 5% NFDM/TBST.
All lanes: Immunoprecipitation - Anti-Histone H3 (citrulline R8) antibody [EPR20358-13] (Anti-Histone H3 (citrulline R8) antibody [EPR20358-13] ab219406)
Predicted band size: 15 kDa
Exposure time: 1s
Flow cytometric analysis of 4% paraformaldehyde-fixed HEK-293T (Human epithelial cell line from embryonic kidney) transfected with empty vector (left panel) or PADI4 (WT, right panel), then treated with 10mM CaCl2 and 10μM Ionomycin (ab120116) for 2 hours, labeling Histone H3 (citrulline R8) with Anti-Histone H3 (citrulline R8) antibody [EPR20358-13] ab219406 at 1/500 dilution, followed by Goat anti rabbit IgG (Alexa Fluor® 647) at 1/2000 dilution.
Positive signal is obtained from HEK-293T cells transfected with WT PADI4 treated with 10mM CaCl2 and 10μM Ionomycin (ab120116) for 2 hours.
Histone H3 (citrulline R17) was immunoprecipitated from 0.35 mg of HEK-293T (Human epithelial cell line from embryonic kidney) transfected with PADI4 (WT), then treated with 10mM CaCl2 and 10μM Ionomycin (ab120116) for 2 hours, whole cell lysate with Anti-Histone H3 (citrulline R17) antibody [EPR20358-120] ab219407 at 1/30 dilution.
Western blot was performed from the immunoprecipitate using Anti-Histone H3 (citrulline R17) antibody [EPR20358-120] ab219407 at 1/1000 dilution.
VeriBlot for IP secondary antibody (HRP) (VeriBlot for IP Detection Reagent (HRP) ab131366), was used for detection at 1/10000 dilution.
Lane 1: HEK-293T transfected with PADI4 (WT), then treated with 10mM CaCl2 and 10μM Ionomycin (ab120116) for 2 hours, whole cell lysate 10 μg (Input).
Lane 2: Anti-Histone H3 (citrulline R17) antibody [EPR20358-120] ab219407 IP in HEK-293T transfected with PADI4 (WT), then treated with 10mM CaCl2 and 10μM Ionomycin (ab120116) for 2 hours, whole cell lysate.
Lane 3: Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) instead of Anti-Histone H3 (citrulline R17) antibody [EPR20358-120] ab219407 in HEK-293T transfected with PADI4 (WT), then treated with 10mM CaCl2 and 10μM Ionomycin (ab120116) for 2 hours, whole cell lysate.
Blocking/Dilution buffer: 5% NFDM/TBST.
All lanes: Immunoprecipitation - Anti-Histone H3 (citrulline R17) antibody [EPR20358-120] (Anti-Histone H3 (citrulline R17) antibody [EPR20358-120] ab219407)
Predicted band size: 15 kDa
Exposure time: 1s
Flow cytometric analysis of 4% paraformaldehyde-fixed HEK-293T (Human epithelial cell line from embryonic kidney) transfected with empty vector (left panel) or PADI4 (WT, right panel), then treated with 10mM CaCl2 and 10μM Ionomycin (ab120116) for 2 hours, labeling Histone H3 (citrulline R17) with Anti-Histone H3 (citrulline R17) antibody [EPR20358-120] ab219407 at 1/500 dilution, followed by Goat anti rabbit IgG (Alexa Fluor® 647) at 1/2000 dilution.
Positive signal is obtained from HEK-293T cells transfected with WT PADI4 treated with 10mM CaCl2 and 10μM Ionomycin (ab120116) for 2 hours.
ab58668 staining ATF3 in A549 cells treated with ionomycin Ca2+ salt (ab120116), by ICC/IF. Increase in ATF3 expression correlates with increased concentration of ionomycin Ca2+ salt, as described in literature.
The cells were incubated at 37°C for 2h in media containing different concentrations of ab120116 (ionomycin Ca2+ salt) in DMSO, fixed with 4% formaldehyde for 10 minutes at room temperature and blocked with PBS containing 10% goat serum, 0.3 M glycine, 1% BSA and 0.1% tween for 2h at room temperature. Staining of the treated cells with ab58668 (10 μg/ml) was performed overnight at 4°C in PBS containing 1% BSA and 0.1% tween. A DyLight 488 goat anti-mouse polyclonal antibody (Goat Anti-Mouse IgG H&L (DyLight® 488) preadsorbed ab96879) at 1/250 dilution was used as the secondary antibody. Nuclei were counterstained with DAPI and are shown in blue.
Jurkat were stimulated for 48 hours with 50 ng x mL-1 of PMA (Phorbol 12-myristate 13-acetate (PMA), PKC activator ab120297) and 1 uM Ionomycin (ab120116) and PBMCs were stimulated for 48 hours with 2 % PHA-M (LifeTechnologies). Cell free supernatants were tested, showing results after background signal was subtracted (duplicates +/- SD).
Blocking and diluting buffer and concentration: 5% NFDM/TBST.
The expression profile/ molecular weight observed is consistent with what has been described in the literature (PMID:23129404 and PMID: 6423641).
In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (Anti-GAPDH antibody [EPR16891] - Loading Control ab181602) staining at 1/200000 dilution.
All lanes: Western blot - Anti-Interferon gamma antibody [RM1245] (Anti-Interferon gamma antibody [RM1245] ab322926) at 1/1000 dilution
Lane 1: Untreated No-GFP-CD16.NK-92 (CD16 ectopically expressed natural killer(NK-92®) cell with no GFP tag) whole cell lysate at 20 µg
Lane 2: No-GFP-CD16.NK-92 treated with 80nM PMA(TPA) (Phorbol 12-myristate 13-acetate (PMA), PKC activator ab120297) and 3uM Ionomycin (ab120116) for 5 hours, 300 ng/ml BFA was added for last 4 hours whole cell lysate at 20 µg
All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/20000 dilution
Observed band size: 17 kDa, 19 kDa, 23 kDa, 36 kDa
Exposure time: 92s
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