5-Nitro BAPTA

CAS: 124251-83-8 F: C22H23N3O12 W: 521.43

5-Nitro BAPTA is a calcium chelator, combinded with 2-Me-substituted TM ( as a fluorescent moiety), can be used to form

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Bioactivity	5-Nitro BAPTA is a calcium chelator, combinded with 2-Me-substituted TM ( as a fluorescent moiety), can be used to form a red fluorescent probe (CaTM-2 AM), for imaging of cytoplasmic Ca2+ in cultured living cells. 5-Nitro BAPTA is a building block used in the synthesis of Ca2+ specific chelators, Ca2+ buffers, and fluorescent Ca2+ indicators[1][2].
Invitro	5-Nitro BAPTA, designed to a red fluorescent probe for cytoplasmic Ca2+ with strong emission in the long-wavelength region[1].General procedure for fluorescence imaging of cultured Hela cells[1]:1.Plate cells onto a 35-mm poly-L-lysinecoated glass-bottomed dish (Matsunami) in DMEM supplemented with 10% (v/v) fetal bovine serum, 1% penicillin and 1% streptomycin. 2. Remove DMEM, wash the dish with HBSS 3 times, and then add CaTM-2 AM (3 μM) in Hanks’ Balanced Salt Solution (HBSS) containing 0.3% DMSO as a cosolvent. 3. Incubate at 37°C for 30 min, remove medium and wash dishes with HBSS 3 times. The cells can be observed in HBSS.4. Capture fluorescence images with excitation and emission wavelength of 590/610–680 nm.General procedure for fluorescence imaging of slices[1]:1. Incubate slide cultures with 2 mL dye solution at 37 ºC for 40 min. The dye solution is artificial cerebrospinal fluid (aCSF) containing 10 μM CaTM-2 AM, 0.01% Pluronic F-127, and 0.005% Cremophor EL. aCSF consisted of : 126 mM NaCl, 26 mM NaHCO3, 3.5 mM KCl, 1.24 mM NaH2PO4, 1.3 mM MgSO4, 1.2 mM CaCl2, and 10 glucose.2. Wash slieds with aCSF three times and recover in 2 mL aCSF at 37 ºC for 45 min, during which 2 µL of 1 mM Acridine orange was added to the aCSF at time 40 min.3. Transferre slice cultures into a recording chamber heated at 35 ºC and continuously perfused with aCSF at 2 mL/min. 4. Acqure images at 10 frames/s with a Nipkowdisk confocal unit (CSUX-1, Yokogawa Electric, Tokyo, Japan), cooled CCD camera (iXon DU897, Andor, Belfast, UK), a water-immersion objective lens (16×, 0. NA, Nikon, Tokyo, Japan), and image acquisition software (Solis, Andor Technology, Belfast, UK). 5. Set the excitation wavelength to 488 nm (7 mW) and 568 nm (15 mW) for Acridine orange and CaTM-2 with an argon-krypton laser (641-YB-A01; Melles Griot, Carlsbad, CA, USA) and set the emission wavelength to 520-535 nm and 617-673 nm band-pass emission filters, respectively. 6. Analysis data with custom-made software written in Microsoft Visual Basic. 7. Calculate fluorescence change ΔF/F as (Ft-F0)/F0, where Ft is the fluorescence intensity at frame time t, and F0 is the average baseline.
Name	5-Nitro BAPTA
CAS	124251-83-8
Formula	C22H23N3O12
Molar Mass	521.43
Transport	Room temperature in continental US; may vary elsewhere.
Storage	Please store the product under the recommended conditions in the Certificate of Analysis.
Reference	[1]. Takahiro Egawa, et al. Red Fluorescent Probe for Monitoring the Dynamics of Cytoplasmic Calcium Ions†. Angew Chem Int Ed. 2013, 52(14):3874-3877. [2]. Jones, et al. Purification and labeling of extracellular vesicles using a mixed mode resin composition: World Intellectual Property Organization, WO2019133842[P]. 2019-07-04.

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