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  5. Invitrogen™ BackDrop™ Background Suppressor for Microplate Assays

Invitrogen™ BackDrop™ Background Suppressor for Microplate Assays

Catalog No. p-4926563
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One 50 mL bottle
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B10512 One 50 mL bottle
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Catalog No. B10512 Supplier Invitrogen™ Supplier No. B10512
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Description

Designed to effectively suppress background fluorescence in live cell assays.

BackDrop Background Suppressor is a novel signal enhancer reagent that helps improve signal-to-noise ratios when used with green-emitting fluorescent dyes or proteins.

  • Can be added to complete media
  • Improves sensitivity of green-emitting fluorescent dyes or proteins
  • Validated with many cellular assays, including ion indicator and membrane potential assays
  • Minimal cell toxicity—demonstrated to not cause osmotic shock to neuronal cells

The sensitivity of fluorescent assays can be compromised by autofluorescence or background fluorescent signals. These background signals result from cellular proteins or growth media components and contribute to a decrease in the signal-to-noise ratio and reduced assay sensitivity. Addition of BackDrop Background Suppressor significantly reduce background signal, helping improve experimental results. BackDrop Background Suppressor can be added to growth media and has been formulated to not contribute to cell toxicity. It has been successfully used with many different cell types and fluorescent reagents (including ion indicators) to efficiently suppress background fluorescence.

Cell Analysis, Cellular Imaging, Immunocytochemistry (ICC)

Specifications

For Use With (Equipment) Fluorescence Microscope
Includes One 50 mL bottle
Quantity 50 mL
Content And Storage Store at 2-8°C. DO NOT FREEZE. Protect material from long-term exposure to light.
Form Liquid
Product Line BackDrop
For Use With (Application) Microplate Assays
Product Type Background Suppressor for live cell assays

Frequently Asked Questions (FAQs)

How do I quench any fluorescence outside of my live cells and leave my intracellular staining intact?

There are a number of reagents used to quench fluorescence on the outside of live cells or in the media (background fluorescence). Our Backdrop Background Suppressor reagents are a great option to quench blue, green, and red fluorescent dyes, various media components and fluorescent proteins. Another commonly used reagent is the cell impermeant colorimetric dye, Trypan blue.

The BackDrop background suppressor solutions are supposed to reduce background fluorescence. Does that include sample autofluorescence?

No. The BackDrop dyes will quench any fluorescence of a given wavelength in the extracellular media or buffer, but not inside live cells. It will not quench autofluorescence inside live cells unless the dye is engulfed.
Can the Fluo-4 Calcium Imaging Kit (Cat. No. F10489) components be purchased separately?

Yes. The following components are sold separately:
Component B: PowerLoad Concentrate (Cat. No. P10020)
Component C: BackDrop Background Suppressor for Microplate Assays (Cat. No. B10512)
Component D: Probenecid, Water Soluble (Cat. No. P36400)
Can I use the BackDrop Background Suppressor reagent as the quencher to reach the inner leaflet of the plasma membrane?

We have not tested this. The BackDrop Background Suppressor reagent is cell impermeant, but if the quenching is caused by a FRET interaction and, if the distance between the BackDrop Background Suppressor outside of the cell is within the Ro distance for FRET with the fluorescent dye in the inner leaflet of the membrane, quenching may still occur.

Would the BackDrop Background Suppressor reagent quench Alexa Fluor 647 signal? What is the mechanism of quenching?

Yes, the BackDrop Background Suppressor reagent can be used to quench Alexa Fluor 647 signal. We have not determined the exact mechanism for the quenching but it depends upon the dye. For some dyes, it may be a FRET interaction, for others the BackDrop Background Suppressor absorbs the photons emitted by the fluorescent dye (inner filter effect).

I understand that some media are worse than others for fluorescence imaging. How do I choose?

Most media contain phenol red, which can quench fluorescent dyes in the visible wavelengths. Most media also contain autofluorescent components, such as riboflavin, which can reduce signal-to-background. We offer FluoroBrite DMEM and HEPES-based Live Cell Imaging Solution, which have been optimized for fluorescent imaging. We also offer a number of media without phenol red. But if none of these are reasonable options for your experiment, then we also offer BackDrop Background Suppressor ReadyProbes Reagent, which can be added to quench media autofluorescence.

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