X rhod 1 am

Manufactured by Thermo Fisher Scientific

Sourced in United States

X-Rhod-1 AM is a fluorescent calcium indicator. It is used to detect and quantify calcium levels in live cells.

Automatically generated - may contain errors

Lab products found in correlation

Close spelling variants of this product

X-Rhod-1 (15 citations)

18 protocols using x rhod 1 am

Fluorescent Probes for Multidrug Resistance

Cited 1 time

Check if the same lab product or an alternative is used in the 5 most similar protocols

The fluorescent compounds tetramethylrosamine chloride (TMR-Cl), BODIPY (BD)-verapamil, 3-propionyl ethylenediamine hydrochloride (BD-EDA), BD-prazosin, BD-vinblastine, tetramethylrhodamine ethyl ester perchlorate (TMRE), 3,3'-diethyloxacarbocyanine iodide (DiOC₂), quinolinium 6-(dimethylamino)-2-[4-[4-(dimethylamino)phenyl]-1,3-butadienyl]-1-ethyl perchlorate (LDS-751), SYTO13, rhodamine 123, calcein-AM, dihydrorhodamine 123, Mito-tracker deep red FM, ER tracker red, Rhod-2-AM, X-Rhod-1-AM, tetramethylrhodamine methyl ester perchlorate (TMRM), rhodamine B hexyl ester were purchased from Invitrogen/ThermoFisher (Carlsbad, CA). Flutax-1 was purchased from Tocris biosciences (Minneapolis, MN). Cyclosporine A was purchased from the Alexis Corporation (Lausen, Switzerland). NBD-cyclosporine A was generously provided by Drs. Anika Hartz and Björn Bauer, University of Kentucky (Lexington, KY). All remaining chemicals were purchased from Sigma-Aldrich (St. Louis, MO).
The P-gp specific monoclonal antibody C219 was provided by Fujirebio Diagnostic Inc. (Malvern, PA), and the MRK16 antibody was purchased from Kyowa Medex Company (Tokyo, Japan). 4E3 antibody was purchased from Abcam (Cambridge, MA) and UIC2 was purchased from eBioscience (San Diego, CA) [17 (link)]. FITC-labeled anti-mouse secondary antibody IgG2aκ isotype was obtained from BD Biosciences (San Jose, CA).

Sajid A., Lusvarghi S., Chufan E.E, & Ambudkar S.V. (2018). Evidence for the critical role of transmembrane helices 1 and 7 in substrate transport by human P-glycoprotein (ABCB1). PLoS ONE, 13(9), e0204693.

+ Open protocol

+ Expand

Fluorescent Probes for P-Glycoprotein Study

Check if the same lab product or an alternative is used in the 5 most similar protocols

The fluorescent substrates BODIPY (BD)-verapamil, BODIPY-3-propionyl ethylenediamine hydrochloride (BD-EDA), BD-prazosin, and tetramethylrhodamine ethyl ester perchlorate (TMRE) were purchased from Serateh Biotech (Eugene, OR, USA). Other fluorescent compounds including 3-quinolinium 6-(dimethylamino)-2-[4-[4-(dimethylamino)-phenyl]-1,3-butadienyl]- 1-ethyl perchlorate (LDS-751), SYTO13, rhodamine 123 (Rh 123), calcein-acetoxymethyl (Cal-AM), Rhod-2-AM, rhodamine-6G (R6G), X-Rhod-1-AM, and rhodamine B hexyl ester (R6) were purchased from Invitrogen/Thermo Fisher (Carlsbad, CA, USA). Flutax-1 was purchased from Tocris Biosciences (Minneapolis, MN, USA). Cyclosporine A was purchased from the Alexis Corporation (Lausen, Switzerland). [N-ε(4-nitro-benzofurazan-7-yl)-D-Lys(8)]-cyclosporine A (NBD-CsA) was generously provided by Drs Anika Hartz and Bjoern Bauer, University of Kentucky (Lexington, KY, USA). All remaining chemicals were obtained from Sigma-Aldrich (St. Louis, MO, USA). The P-gp-specific monoclonal antibody C219 was provided by Fujirebio Diagnostic Inc. (Malvern, PA, USA), the MRK16 antibody was purchased from Kyowa Medex Company (Tokyo, Japan), and UIC2 antibody was purified from hybridoma cells as described in [28 (link)]. FITC-labeled anti-mouse secondary antibody IgG2a κ isotype was obtained from BD Biosciences (San Jose, CA, USA).

Rahman H., Ware M.J., Sajid A., Lusvarghi S., Durell S.R, & Ambudkar S.V. (2023). Residues from Homologous Transmembrane Helices 4 and 10 Are Critical for P-Glycoprotein (ABCB1)-Mediated Drug Transport. Cancers, 15(13), 3459.

+ Open protocol

+ Expand

Mitochondrial Morphology and Viability

Check if the same lab product or an alternative is used in the 5 most similar protocols

Mito Tracker Red CMXRos (C1049B) and Calcein/PI Cell Viability/Cytotoxicity Assay was brought from Beyotime (C2015S). X-rhod-1, AM was purchased from Thermo Fisher (X14210). All these tests were performed according to the kit’s instructions. Morphological analysis of liver mitochondrial by transmission electron microscopy (Servicebio).

Qin D., Wang R., Ji J., Wang D., Lu Y., Cao S., Chen Y., Wang L., Chen X, & Zhang L. (2023). Hepatocyte-specific Sox9 knockout ameliorates acute liver injury by suppressing SHP signaling and improving mitochondrial function. Cell & Bioscience, 13, 159.

+ Open protocol

+ Expand

Measuring Mitochondrial Calcium Uptake

Cited 1 time

Check if the same lab product or an alternative is used in the 5 most similar protocols

Mitochondrial Ca²⁺ uptake was monitored in freshly isolated control and DIC-treated cardiomyocytes using X-Rhod-1 AM (ThermoFisher Scientific) [17 (link)]. Cells were loaded with X-Rhod-1 AM for 40 min at 37 °C. Mitochondrial Ca²⁺ uptake was monitored by the change in fluorescence intensity, normalized to the baseline fluorescence intensity, after the addition of 5 μM Ca²⁺ and 10 μM Ca²⁺.

Thai P.N., Ren L., Xu W., Overton J., Timofeyev V., Nader C.E., Haddad M., Yang J., Gomes A.V., Hammock B.D., Chiamvimonvat N, & Sirish P. (2021). Chronic Diclofenac Exposure Increases Mitochondrial Oxidative Stress, Inflammatory Mediators, and Cardiac Dysfunction. Cardiovascular Drugs and Therapy, 37(1), 25-37.

+ Open protocol

+ Expand

Calcium Imaging of Cells

Check if the same lab product or an alternative is used in the 5 most similar protocols

Cells were incubated with X-Rhod-1AM (Thermo, X14210) at a 1:4,000 dilution for 10 min, washed twice with PBS and the cells were imaged in widefield at ×10 with the cells in serum-free medium. Using our custom MATLAB tracking software we masked every cell using the H2B:GFP signal and measured the calcium inside this area.

Park W.Y., Gray J.M., Holewinski R.J., Andresson T., So J.Y., Carmona-Rivera C., Hollander M.C., Yang H.H., Lee M., Kaplan M.J., Cappell S.D, & Yang L. (2023). Apoptosis-induced nuclear expulsion in tumor cells drives S100a4-mediated metastatic outgrowth through the RAGE pathway. Nature Cancer, 4(3), 419-435.

+ Open protocol

+ Expand

Monitoring Mitochondrial Calcium Uptake

Cited 1 time

Check if the same lab product or an alternative is used in the 5 most similar protocols

Mitochondrial Ca²⁺ Uptake was monitored in freshly isolated control and DIC-treated cardiomyocytes using X-Rhod-1 AM (ThermoFisher Scientific) [17 (link)]. Cells were loaded with X-Rhod-1 AM for 40 minutes at 37°C. Mitochondrial Ca²⁺ uptake was monitored by the change in fluorescence intensity, normalized to the baseline fluorescence intensity, after the addition of 5 μM Ca²⁺ and 10 μM Ca²⁺.

+ Open protocol

+ Expand

Analyzing Intracellular Ca2+ Transients in hESC/iPSC-derived Cardiomyocytes

Check if the same lab product or an alternative is used in the 5 most similar protocols

Intracellular Ca²⁺ transients were analyzed by loading hESC/iPSC-vCMs with 2.5 μmol/L X-Rhod-1 AM (Invitrogen) for 45 minutes at 37°C in DMEM/F12, followed by imaging with a complementary metal-oxide semiconductor–based camera (MiCAM ULTIMA, SciMedia USA Ltd, CA) in Tyrode’s solution containing 140 mmol/L NaCl, 5 mmol/L KCl, 1 mmol/L CaCl₂, 1 mmol/L MgCl₂, 10 mmol/L glucose, 10 mmol/L HEPES, and pH adjusted to 7.4 with NaOH. Electric stimulation or 50 mmol/L caffeine was applied to evoke Ca²⁺ transients. Isoproterenol (1 μmol/L) was incubated with hESC/iPSC-vCMs for 3 to 5 minutes at 37°C before measurement; 10 μmol/L ryanodine or 1 μmol/L thapsigargin was incubated with hES2-vCMs, as indicated, for 30 minutes at 37°C before measurements.

Chen G., Li S., Karakikes I., Ren L., Chow M.Z., Chopra A., Keung W., Yan B., Chan C.W., Costa K.D., Kong C.W., Hajjar R.J., Chen C.S, & Li R.A. (2014). Phospholamban as a Crucial Determinant of the Inotropic Response of Human Pluripotent Stem Cell–Derived Ventricular Cardiomyocytes and Engineered 3-Dimensional Tissue Constructs. Circulation. Arrhythmia and electrophysiology, 8(1), 193-202.

+ Open protocol

+ Expand

Lysosomal and Cytosolic Calcium Imaging

Check if the same lab product or an alternative is used in the 5 most similar protocols

For lysosomal calcium measurements, we performed Oregon Green^® 488 BAPTA-1 dextran (OGBD) labeling^{41 (link)}. In this assay, cells with a stable transduction of LAMP1-mRFP lentiviruses were loaded with 0.1 mg ml⁻¹ OGBD (Invitrogen) for 30 min. For cytosolic calcium measurements, we performed X-Rhod-1 and Fura-2 labeling assays. In these assays, cells were loaded with 1 μM X-Rhod-1-AM (Invitrogen) or 1 μM Fura-2-AM (Invitrogen) for 30 min. For OGBD and X-Rhod-1 imaging, the cells were examined under a temperature- and CO₂-controlled laser confocal microscope (Olympus); for ratiometric Fura-2 imaging, cells were examined under a temperature- and CO₂-controlled fluorescence microscope equipped with excitation wavelengths of 340 and 380 nm (Till Polychrome V, Till Photonics, Gräfelfing, Germany), a 410-dLcp beamsplitter, a D510/80 wide band emission filter (Chroma Technology Corp., Rockingham, VT, USA) and a Quant-EM camera (Photometrics, Tucson, AZ, USA).

Park H.W., Park H., Semple I.A., Jang I., Ro S.H., Kim M., Cazares V.A., Stuenkel E.L., Kim J.J., Kim J.S, & Lee J.H. (2014). Pharmacological correction of obesity-induced autophagy arrest using calcium channel blockers. Nature communications, 5, 4834.

+ Open protocol

+ Expand

Measurement of Cytosolic and Mitochondrial Calcium

Check if the same lab product or an alternative is used in the 5 most similar protocols

CTRL and Pt primary fibroblasts were cultured onto 24 × 24 mm glass slides in six-well plates and incubated for 30 min at 37 °C, with the following probes: 2 µM Fluo-4 AM (Life Technologies, Carlsbad, CA, USA) for cytosolic Ca²⁺ and 4 µM X-Rhod-1 AM (Invitrogen, Molecular probes^TM, Carlsbad, CA, USA) for mitochondrial Ca²⁺ level measurements. Stained cells were washed with Phosphate Buffered Saline (PBS), fixed in PBS containing 4% paraformaldehyde for 10 min at room temperature, and cell nuclei were stained with 1 μg/mL 4′,6-Diamidino-2-phenyindole, dilactate (DAPI) Sigma-Aldrich, St. Louis, MO, USA) for 10 min at room temperature. Stained cells were examined with a Leica TCS SP8 microscope (images collected using a 40× and 60× in oil immersion objective) coupled to the Laser Scanning Confocal Microscopy (LSCM) system. Acquisition, storage, and data analysis were performed using Leica software LAS AF 3 (https://www.leica-microsystems.com/products/microscope-software/).

Tanzarella P., Ferretta A., Barile S.N., Ancona M., De Rasmo D., Signorile A., Papa S., Capitanio N., Pacelli C, & Cocco T. (2019). Increased Levels of cAMP by the Calcium-Dependent Activation of Soluble Adenylyl Cyclase in Parkin-Mutant Fibroblasts. Cells, 8(3), 250.

+ Open protocol

+ Expand

Analysis of T-cell signaling cascades

Check if the same lab product or an alternative is used in the 5 most similar protocols

Anti-CD3 (clone OKT3), anti-ZAP-70 (clone 1E7.2), mouse IgG1 isotype control, and APC-conjugated anti-mouse IgG1 (clone M1-14D12) were from eBioscience. Rabbit anti-GFP (clone ab290) and goat anti-Rabbit IgG H&L (HRP) (clone ab6721) were from abcam. X-rhod-1-AM, probenecid, Dynabeads, and anti-β tublin antibody were from Invitrogen. Goat anti-mouse IgG peroxidase conjugated, and mouse anti-goat IgG H&L were from Thermo Scientifc. Rabbit anti-phosphoZAP-70 was from Cell Signaling. Mouse anti-phosphotyrosine 4G10 was from Millipore. Superantigen was a mix of recombinant staphylococcal enterotoxin E, staphylococcal enterotoxin A, staphylococcal enterotoxin B, and staphylococcal enterotoxin C3 coming from Toxin Technology. All chemicals and other reagents were from Sigma unless otherwise indicated.

Li K., Xiang X., Sun J., He H.T., Wu J., Wang Y, & Zhu C. (2016). Imaging Spatiotemporal Activities of ZAP-70 in Live T Cells Using a FRET-based Biosensor. Annals of biomedical engineering, 44(12), 3510-3521.

+ Open protocol

+ Expand

About PubCompare

Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.

We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.

However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.

Ready to get started?

Revolutionizing how scientists
search and build protocols!