Ionomycin Ca2+ Salt, Ca2+ ionophore (ab120116)
Key features and details
- Ca2+ ionophore
- CAS Number: 56092-82-1
- Soluble in DMSO to 25 mM and in ethanol to 100 mM
- Form / State: Solid
- Source: Streptomyces conglobatus
Overview
-
Product name
Ionomycin Ca2+ Salt, Ca2+ ionophore -
Description
Ca2+ ionophore -
CAS Number
56092-82-1 -
Chemical structure
Properties
-
Chemical name
(4R,6S,8S,10Z,12R,14R,16E,18R,19R,20S,21S)-11,19,21-Trihydroxy-4,6,8,12,14,18,20-heptamethyl-22-[(2S,2'R,5S,5'S)-octahydro-5'-[(1R)-1-hydroxyethyl]-2,5'-dimethyl[2,2'-bifuran]-5-yl]-9-oxo-10,16-docosadienoic acid calcium salt -
Molecular weight
747.08 -
Molecular formula
C41H70CaO9 -
PubChem identifier
6446270 -
Storage instructions
Store at +4°C. Store under desiccating conditions. The product can be stored for up to 12 months. -
Solubility overview
Soluble in DMSO to 25 mM and in ethanol to 100 mM -
Handling
Wherever possible, you should prepare and use solutions on the same day. However, if you need to make up stock solutions in advance, we recommend that you store the solution as aliquots in tightly sealed vials at -20°C. Generally, these will be useable for up to one month. Before use, and prior to opening the vial we recommend that you allow your product to equilibrate to room temperature for at least 1 hour.
Need more advice on solubility, usage and handling? Please visit our frequently asked questions (FAQ) page for more details.
-
SMILES
[Ca+2].[O-]C(=O)CC[C@@H](C)C[C@H](C)C[C@H](C)C(=O)C=C(/[O-])[C@H](C)C[C@H](C)CC=C[C@@H](C)[C@@H](O)[C@@H](C)[C@@H](O)C[C@@H]1CC[C@](C)(O1)[C@H]2CC[C@](C)(O2)[C@@H](C)O -
Source
Streptomyces conglobatus
-
Research areas
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab120116 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
| Application | Abreviews | Notes |
|---|---|---|
| Functional Studies |
Use at an assay dependent concentration.
|
| Notes |
|---|
|
Functional Studies
Use at an assay dependent concentration. |
Images
-
Chemical Structure - Ionomycin Ca2+ Salt, Ca2+ ionophore (ab120116)2D chemical structure image of ab120116, Ionomycin Ca2+ Salt, Ca2+ ionophore
-
Western blot - Ionomycin Ca2+ Salt, Ca2+ ionophore (ab120116)All lanes : Anti-IL-17A antibody [EPR21776] (ab218013) at 1/1000 dilution
Lane 1 : Untreated EL4 (mouse lymphoma T lymphocyte) whole cell lysate
Lane 2 : EL4 treated with 50 ng/ml Phorbol-12-myristate-13-acetate (PMA) and 500 ng/ml ionomycin (ab120116) for 6 hours, then with 500 ng/ml Brefeldin A for 18 hours, whole cell lysate
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution
Developed using the ECL technique.
Observed band size: 14,17 kDa why is the actual band size different from the predicted?
Exposure time: 3 minutesBlocking/dilution buffer and concentration: 5% NFDM/TBST
Expression of IL-17A can be induced by PMA and Ionomycin treatment (PMID 28382171).
The expression profile is consistent with the literature (PMID 9764847).
-
Western blot - Ionomycin Ca2+ Salt, Ca2+ ionophore (ab120116)All lanes : Anti-Histone H3 (citrulline R8) antibody [EPR20358-13] (ab219406) at 1/10000 dilution
Lane 1 : Whole cell lysate from HEK-293T (Human epithelial cell line from embryonic kidney) transfected with empty vector with GFP tag (vector control), then treated with 10mM CaCl2 and 10µM Ionomycin (ab120116) for 2 hours
Lane 2 : Whole cell lysate from HEK-293T cells transfected with PADI4 (WT), then treated with 10mM CaCl2 and 10µM Ionomycin (ab120116) for 2 hours
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution
Observed band size: 15 kDa why is the actual band size different from the predicted?
Exposure time: 1 secondBlocking/Dilution buffer: 5% BSA/TBST.
Histone H3R8 is citrullinated by PADI4 and CaCl2 is used as a cofactor according to the literature (PMID: 16567635). Ionomycin is used to improve the modification by PADI4 according to the literature (PMID: 26360112).
-
Western blot - Ionomycin Ca2+ Salt, Ca2+ ionophore (ab120116)All lanes : Anti-Histone H3 (citrulline R8) antibody [EPR20358-13] (ab219406) at 1/5000 dilution
Lane 1 : Whole cell lysate from NIH/3T3 (Mouse embryonic fibroblast cell line) transfected with empty vector with GFP tag (vector control), then treated with 10mM CaCl2 for 2 hours
Lane 2 : Whole cell lysate from NIH/3T3 transfected with PADI4 (WT) then treated with 10mM CaCl2 for 2 hours
Lane 3 : Whole cell lysate from C6 (Rat glial tumor cell line) transfected with empty vector with GFP tag (vector control) then treated with 10mM CaCl2 and 10µM Ionomycin (ab120116) for 2 hours
Lane 4 : C6 transfected with PADI4 (WT), then treated with 10mM CaCl2 and 10µM Ionomycin (ab120116) for 2 hours
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution
Observed band size: 15 kDa why is the actual band size different from the predicted?
Exposure time: 1 secondBlocking/Dilution buffer: 5% BSA/TBST.
-
Flow Cytometry (Intracellular) - Ionomycin Ca2+ Salt, Ca2+ ionophore (ab120116)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 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.
-
Immunoprecipitation - Ionomycin Ca2+ Salt, Ca2+ ionophore (ab120116)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 ab219406 at 1/30 dilution.
Western blot was performed from the immunoprecipitate using ab219406 at 1/1000 dilution.
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: 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 (ab172730) instead of 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.
-
Western blot - Ionomycin Ca2+ Salt, Ca2+ ionophore (ab120116)All lanes : Anti-Histone H3 (citrulline R17) antibody [EPR20358-120] (ab219407) at 1/5000 dilution
Lane 1 : HEK-293T (Human epithelial cell line from embryonic kidney) transfected with empty vector with GFP tag (vector control), then treated with 10mM CaCl2 and 10µM Ionomycin (ab120116) for 2 hours, whole cell lysate
Lane 2 : 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
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution
Observed band size: 15 kDa why is the actual band size different from the predicted?
Exposure time: 3 secondsBlocking/Dilution buffer: 5% BSA/TBST.
Histone H3R17 is citrullinated by PADI4 and CaCl2 is used as a cofactor according to the literature (PMID: 16567635). Ionomycin is used to improve the modification by PADI4 according to the literature (PMID: 26360112).
-
Western blot - Ionomycin Ca2+ Salt, Ca2+ ionophore (ab120116)All lanes : Anti-Histone H3 (citrulline R17) antibody [EPR20358-120] (ab219407) at 1/5000 dilution
Lane 1 : NIH/3T3 (Mouse embryonic fibroblast cell line) transfected with empty vector with GFP tag (vector control), then treated with 10mM CaCl2 for 2 hours, whole cell lysate
Lane 2 : NIH/3T3 (Mouse embryonic fibroblast cell line) transfected with PADI4 (WT) then treated with 10mM CaCl2 for 2 hours, whole cell lysate
Lane 3 : C6 (Rat glial tumor cell line) transfected with empty vector with GFP tag (vector control) then treated with 10mM CaCl2 and 10µM Ionomycin (ab120116) for 2 hours, whole cell lysate
Lane 4 : C6 (Rat glial tumor cell line) transfected with PADI4 (WT), then treated with 10mM CaCl2 and 10µM Ionomycin (ab120116) for 2 hours, whole cell lysate
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution
Observed band size: 15 kDa why is the actual band size different from the predicted?
Exposure time: 1 secondBlocking/Dilution buffer: 5% BSA/TBST.
-
Flow Cytometry (Intracellular) - Ionomycin Ca2+ Salt, Ca2+ ionophore (ab120116)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 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.
-
Immunoprecipitation - Ionomycin Ca2+ Salt, Ca2+ ionophore (ab120116)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 ab219407 at 1/30 dilution.
Western blot was performed from the immunoprecipitate using ab219407 at 1/1000 dilution.
VeriBlot for IP secondary antibody (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: 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 (ab172730) instead of 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.
-
Functional Studies - Ionomycin Ca2+ Salt, Ca2+ ionophore (ab120116)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 (ab96879) at 1/250 dilution was used as the secondary antibody. Nuclei were counterstained with DAPI and are shown in blue. -
Sandwich ELISA - Ionomycin Ca2+ Salt, Ca2+ ionophore (ab120116)Sandwich ELISA - IFN gamma Human ELISA Kit (ab46025)
Jurkat were stimulated for 48 hours with 50 ng x mL-1 of PMA (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).
Protocols
To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.
Datasheets and documents
-
SDS download
-
Datasheet download
References (6)
ab120116 has been referenced in 6 publications.
- Dissanayake K et al. Potential applicability of cytokines as biomarkers of disease activity in rheumatoid arthritis: Enzyme-linked immunosorbent spot assay-based evaluation of TNF-a, IL-1ß, IL-10 and IL-17A. PLoS One 16:e0246111 (2021). PubMed: 33497394
- Wang Z et al. Autophagy-based unconventional secretion of HMGB1 by keratinocytes plays a pivotal role in psoriatic skin in?ammation. Autophagy 17:529-552 (2021). PubMed: 32019420
- Waldeck-Weiermair M et al. Rearrangement of MICU1 multimers for activation of MCU is solely controlled by cytosolic Ca(2.). Sci Rep 5:15602 (2015). PubMed: 26489515
- Redmond WJ et al. Nordihydroguaiaretic acid activates hTRPA1 and modulates behavioral responses to noxious cold in mice. Pharmacol Res Perspect 2:e00079 (2014). PubMed: 25505619
- Pont JN et al. Oxytocin-stimulated NFAT transcriptional activation in human myometrial cells. Mol Endocrinol 26:1743-56 (2012). PubMed: 22902539
- Ogawa A et al. PDGF enhances store-operated Ca2+ entry by upregulating STIM1/Orai1 via activation of Akt/mTOR in human pulmonary arterial smooth muscle cells. Am J Physiol Cell Physiol 302:C405-11 (2012). PubMed: 22031597