Apotracker™ Tetra Alexa Fluor® 647

Kit Contents

Kit Contents

For Cat# 427405

• 20 µL Apo-Monomer solution
• 24 µL Alexa Fluor® 647 Streptavidin solution

For Cat# 427406

• 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

Documentation

• Technical Data Sheet (pdf)
• Certificate of Analysis
• Safety Data Sheet

Reviews

Related Protocols

• Cell Surface Flow Cytometry Staining Protocol

Related Pages & Pathways

Related FAQs

Certificate of Analysis