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Invitrogen™ Fura Red™, AM, cell permeant

Catalog No. F3020
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10 x 50 μg
500 μg
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F3020 500 μg
F3021 10 x 50 μg
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Catalog No. F3020 Supplier Invitrogen™ Supplier No. F3020
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Description

Visible light-excitable fura-2 analog that offers particular possibilities for ratiometric measurement of Ca2+ in single cells

  • Measure by microphotometry, imaging or flow cytometry when used with single excitation, green-fluorescent calcium indicator
  • Acetoxymethyl (AM) ester form
  • Useful for noninvasive intracellular loading

Calcium Detection, Cell Analysis, Cell Viability, Proliferation and Function, Cell-Based Second Messenger Assays, Drug Discovery and Development, G-Protein Coupled Receptor Biology, Ionic Homeostasis and Signaling, Target and Lead Identification and Validation

Order Info

Shipping Condition: Room temperature

Specifications

Content And Storage Store in freezer -5°C to -30°C and protect from light.
Detection Method Fluorescence
For Use With (Application) Cell Viability and Proliferation
For Use With (Equipment) Fluorescence Microscope, Microphotometer, Flow Cytometer
Product Type Calcium Indicator
Dye Type Fluorescent Dye-Based
Product Line Fura Red
Quantity 500 μg
Shipping Condition Room Temperature

Frequently Asked Questions (FAQs)

I am doing calcium flux imaging with your Fura-2 calibration kit, but am seeing a large variability in ratio in different places around the slide. I am correcting for uniform illumination, using the product as directed, and sealing the coverslip with nail polish.

The nail polish may be the problem. The Kd value (calcium sensitivity) changes depending upon the dye's environment. Nail polish has solvents that can leech under the coverslip and cause variability. We recommend either going without a sealing or sealing with melted paraffin painted on the coverslip edges with a cotton-tipped applicator (paraffin is hydrophobic and has no solvents).
I need to label cells with Fluo-4, AM, for a calcium flux assay. How long after labeling will the dye be retained?

After loading dye into the cells, intracellular esterases remove the 'AM' moiety from the dye. When the 'AM' group is removed, the dye is able to bind calcium and fluoresce. Since the dye is not covalently bound to any cellular components, it may be actively effluxed from the cell. The rate of efflux is dependent upon the inherent properties of the cell, culture conditions and other factors. The dye may be retained for hours, days or even weeks or lost in a matter of minutes. The use of Probenecid (Cat. No. P36400) limits loss by active efflux.

Where can I find the manual for the Fura Red, AM, cell permeant (Cat. No. F3020, F3021)?

Unfortunately, we do not have a user manual for the Fura Red, AM, cell permeant (Cat. No. F3021). You can reference section 19.3 of the Molecular Probes handbook for recommendations for use.
There is also a large number of citations & references provided on the product page.

Basically, you can dissolve the product in good quality anhydrous DMSO to prepare a stock solution of 1-10 mM and use a final solution of about 1-10 µM in a physiological buffer. You can then incubate for 15 to 60 minutes, wash and then leave the cells for about 30 minutes to allow the AM group to hydrolyse before starting the assay. You can add Pluronic F-127 (e.g., Cat. No. P6866) to facilitate uptake. Store the DMSO stock solution at -20 degrees C.

For Research Use Only. Not for use in diagnostic procedures.