ReadyProbes® Ready-to-Use Imaging Reagents  

ReadyProbes® reagents are ready-to-use imaging solutions designed to allow you to stain cells without the need for calculations, dilutions, or pipetting. Simply add 2 drops/mL from the convenient dropper bottle, and you’re ready for the next step. We’ve developed nuclear and actin stains, cell viability and apoptosis stains, antibody conjugates, and imaging accessories—and many of these user-friendly formulations can be kept at room temperature, right on your lab bench.

Nuclear stains

Nuclear stains protocol

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1. Add 2 drops per mL of media

clock 5

 

2. Incubate 5–20 min at 20–25ºC

 

3. Image cells

Sample image

NucRed® Live 647 ReadyProbes® reagent

Nuclear staining. hUVEC cells were grown in a MatTek glass-bottom dish overnight then stained with NucRed® Live 647 reagent.
ReadyProbes® reagent Storage Filter Ex/Em Cat. No.
NucBlue® Live Cell Stain Room temperature (RT) (≤25°C) DAPI 358/461 R37605
NucBlue® Fixed Cell Stain RT (≤25°C) DAPI 352/461 R37606
Propidium Iodide Reagent RT (≤25°C) RFP 535/617 R37108
NucRed® Live 647 Cell Stain RT (≤25°C) Cy®5 638/686 R37106

Actin stains

Actin stains protocol

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1. Apply to fixed and permeabilized cells using 2 drops per mL of media

clock 5


2. Incubate 30 min


3. Wash with PBS, HBSS, or LCIS


4. Image cells

Sample image

Actin staining with the Image-iT® Fixation/Permeabilization Kit

Actin staining. BPAE cells were fixed, permeabilized and blocked using the Image-iT® Fixation/Permeabilization Kit. Tubulin was labeled using an anti-tubulin antibody followed by detection with an Alexa Fluor® 594 secondary. Actin was stained using ActinGreen™ 488 ReadyProbes® reagent and nuclei were stained with NucBlue® Fixed Cell Stain.
ReadyProbes™ reagent Storage Filter Ex/Em Cat. No.
ActinGreen™ 488 ReagentRoom Temperature (RT) (≤25°C)FITC495/518R37110
ActinRed™ 555 ReagentRT (≤25°C)RFP540/565R37112

Viability stains

Viability stains protocol

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1. Add 2 drops per mL of media

clock 5

 

2. Incubate 5–20 min at 20–25ºC

 

3. Image cells

Sample image

Jurkat cell viability determination using the ReadyProbes® Cell Viability Imaging Kit (Blue/Red)

Cell viability determination. Live Jurkat cells or Jurkat cells treated with 0.1% Triton® X-100 detergent were mixed and stained with the reagents in the ReadyProbes® Cell Viability Imaging Kit (Blue/Red)NucBlue® Live stain and propidium iodide (2 drops per milliliter of cell suspension) for 15 minutes. Cells were then imaged and analyzed on the EVOS® FL Auto system. NucBlue® Live stains all cells, while propidium iodide stains only dead cells.
ReadyProbes® reagent Storage Filter Ex/Em Cat. No.
NucGreen® Dead 488 Reagent Room temperature (RT) (≤25°C) FITC 504/523 R37109
NucRed® Dead 647 Cell Stain RT (≤25°C) Cy®5
642/661 R37113
ReadyProbes® Cell Viability Imaging Kit (Blue/Green RT (≤25°C) DAPI/FITC 360/460
504/523
R37609
ReadyProbes® Cell Viability Imaging Kit (Blue/Red) RT (≤25°C) DAPI/RFP 360/460
535/617
R37610

Apoptosis reagents

Apoptosis reagents protocol

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1. Add 2 drops per mL of media

clock 5


2. Incubate 30–60 minutes

 

3. Image cells

Sample image

CellEvent™ Caspase 3/7 Green ReadyProbes™ reagent

Apoptosis determination. HeLa cells were loaded with CellEvent® Caspase 3/7 Green ReadyProbes® reagent, then treated with 0.5 µM staurosporine or vehicle control for 4 hours. Untreated cells (A) have minimal fluorescence, while treated cells (B) have a significant increase in fluorescence (shown in green) indicating apoptosis.
ReadyProbes™ reagent Storage Filter Ex/Em (nm) Cat. No.
CellEvent® Caspase 3/7 Green Room temperature (RT) (≤25°C) FITC 495/518 R37111

Secondary antibodies

Secondary antibodies protocol

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1. Apply 2 drops/mL of solution to fixed and permeabilized cells labeled with primary antibodies

clock 5


2. Incubate 30 minutes at 20–25°C


3. Wash twice with PBS, HBSS, or LCIS (Live Cell Imaging Solution)


4. Image cells

Sample image
Mitochondria detection with Alexa Fluor® 594 goat anti-mouse IgG antibody antibody
Cell staining following fixation and permeabilization. CAKI cells were fixed and permeabilized using the Image-iT® Fixation⁄Permeabilization Kit. The nucleus (blue) is stained with NucBlue® Fixed Cell Stain, actin cytoskeleton (green) is stained with ActinGreen™ 488 Reagent, and mitochondria (red) are labeled with an anti-OxPhos Complex V inhibitor primary antibody that is detected with an Alexa Fluor® 594 goat anti-mouse IgG antibody.

Imaging accessories

Image-iT® Fixation/Permeabilization Kit    
Cell staining following fixation and permeabilization. U2OS cells expressing CellLight® Mitochondria-GFP were fixed and permeabilized with Image-iT® Fixation/Permeabilization Kit. Actin fibers were stained with Alexa Fluor® 594 phalloidin and nuclei with NucBlue® Live Cell Stain.

ReadyProbes® reagent Dropper Storage Live or Fixed Cat. No.
BackDrop® Background Suppressor Yes Room temperature (RT) (≤25°C) Live R37603
Live Cell Imaging Solution No RT (≤25°C) Live A14291DJ
Image-iT® Fixation/Permeabilization Kit No RT (≤25°C) Fixed R37602
Image-iT® FX Signal Enhancer Yes RT (≤25°C) Fixed R37107

Citations

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  • Shikatani EA et al. (2012) Inhibition of proliferation, migration, and proteolysis contribute to corticosterone-mediated inhibition of angiogensis. PLoS One doi:10.1371/journal.pone.0046625.
  • Toyoshima K et al. (2012) Fully functional hair follicle regeneration through the rearrangement of stem cells and their niches. Nature Comm doi:10.1038/ncomms1784.
  • Weng A et al. (2012) Saponins modulate the intracellular trafficking of protein toxins. Jour Contr Release doi: 10.1016/j.jconrel.2012.10.002.
For Research Use Only.  Not for human or animal therapeutic or diagnostic use.