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Apotracker™ Tetra Alexa Fluor® 647

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Regulatory Status
RUO
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Apotracker-Tetra_Alexa-Fluor-647_053123
Heat induced 40% dead C57BL/6 splenocytes stained with Helix NP™ Blue and Apotracker™ Tetra Alexa Fluor® 647 (left) or Alexa Fluor® 647 Streptavidin (right).
  • Apotracker-Tetra_Alexa-Fluor-647_053123
    Heat induced 40% dead C57BL/6 splenocytes stained with Helix NP™ Blue and Apotracker™ Tetra Alexa Fluor® 647 (left) or Alexa Fluor® 647 Streptavidin (right).
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427405 20 tests 76€
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Description

Apoptotic and/or necrotic cells display phosphatidylserine (PS) on their outer cell surface, where it acts as an ‘eat-me’ signal and contributes to efficient removal of dead cells by macrophages and other phagocytes. PS is normally only found on the intracellular leaflet of the plasma membrane in healthy cells, but during early apoptosis, membrane asymmetry is lost and PS translocates to the external leaflet. Apotracker™ monomer binds to PS for the detection and removal of dead or dying cells which are effective and preferably at the same time buffer-independent (Calcium free buffer) in comparison to the agents used in the market thus far.

Product Details
Technical Data Sheet (pdf)

Kit Contents

Kit Contents

100 µL Apo-Monomer solution

120 µL Alexa Fluor® 647 Streptavidin solution

Product Details

Verified Reactivity
Human, Mouse
Formulation
Apo-Monomer: HEPES buffered saline w/ 8% Glycerol, pH 7.4

Streptavidin: Phosphate-buffered solution, pH 7.2, containing 0.09% sodium azide
Preparation
The Apo monomer was purified by affinity chromatography and conjugated with biotin under optimal conditions. Streptavidin was conjugated with Alexa Fluor® 647 under optimal conditions.
Storage & Handling
Kit components should be stored undiluted between 2°C and 8°C, and protected from prolonged exposure to light. Do not freeze.
Application

FC - Quality tested
Recommended Usage

Each lot of this product is quality control tested by immunofluorescent staining with flow cytometric analysis. The suggested use of this reagent is 5 µL of the Apo-Monomer + Streptavidin-Fluorophore mixture per million cells in 100 µL volume.

Please see the detailed protocol in the application notes section below. 

Excitation Laser
Red Laser (633 nm)
Application Notes

Apotracker™ Tetra staining procedure for flow cytometry analysis:

  1. To prepare 25 µL (5 tests) of the tetramer solution, mix 5 µL of Apo-monomer, 6 µL of Fluor-Streptavidin and 14 µL of Cell Staining Buffer (BioLegend Cat. No. 420201), or equivalent. 

    Note: We do not recommend preparing less than 25 µL (5 tests) of the Apo-monomer-Streptavidin complex at a time to avoid pipetting errors. Scale up as necessary
     

  2. For each sample containing 1x106 cells resuspended in 100 µL of Cell Staining Buffer, add 5 µL of tetramer solution prepared in the above step.   
     

  3. Incubate in the dark at room temperature for 15 minutes. 
     

  4. Wash each tube by adding 2 mL of Cell Staining Buffer, or equivalent, and centrifuging at 1200 rpm for five minutes. Remove the supernatant and vortex tubes to loosen pellet. 
     

  5. Repeat wash step above. 
     

  6. Resuspend the pellet by adding 300 - 500 μL Cell Staining Buffer to each tube. 
     

  7. (Optional) Ten minutes prior to flow cytometric analysis, add 10 μL of DAPI solution (or equivalent dye) to each tube for viability staining. 
     

  8. Cells are ready for flow cytometric analysis.

Antigen Details

Gene ID
NA

Related FAQs

There are no FAQs for this product.
Go To Top Version: 2    Revision Date: 07-28-2023

For research use only. Not for diagnostic use. Not for resale. BioLegend will not be held responsible for patent infringement or other violations that may occur with the use of our products.

 

*These products may be covered by one or more Limited Use Label Licenses (see the BioLegend Catalog or our website, www.biolegend.com/ordering#license). BioLegend products may not be transferred to third parties, resold, modified for resale, or used to manufacture commercial products, reverse engineer functionally similar materials, or to provide a service to third parties without written approval of BioLegend. By use of these products you accept the terms and conditions of all applicable Limited Use Label Licenses. Unless otherwise indicated, these products are for research use only and are not intended for human or animal diagnostic, therapeutic or commercial use.

 

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Toll-Free Phone: 1-877-Bio-Legend (246-5343) Phone: (858) 768-5800 Fax: (877) 455-9587

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