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Rat Regulatory T Cell Multi-Color Flow Kit

R&D Systems, part of Bio-Techne | Catalog # FMC015

Contains conjugated antibodies to CD25-PE (Goat IgG), FoxP3-APC (Goat IgG), CD4-Fluorescein (Clone OX-38)
R&D Systems, part of Bio-Techne

Key Product Details

Rat Regulatory T Cell Multi-Color Flow Kit
Discontinued Product
FMC015 has been discontinued. An alternative/replacement product is available: FMC015B. View all Regulatory T Cell Kits products.

Assay Procedure

Refer to the product datasheet for complete product details.

Briefly, rat regulatory T cells are assessed by flow cytometry using the following procedure:

  • Wash and resuspend cells in Flow Cytometry Staining Buffer
  • Stain cells with cell surface marker fluorochrome-conjugated antibodies or isotype controls
  • Wash and resuspend cells in Flow Cytometry FoxP3 Staining Buffer
  • Stain cells with FoxP3-APC antibody or isotype control
  • Wash cells and resuspend in Flow Cytometry Staining Buffer
  • Analyze samples by flow cytometry
 

 

Kit Components

Reagents Supplied in the Rat Regulatory T Cell Multi-Color Flow Cytometry Kit (Catalog # FMC015):

  • CD25-PE (goat IgG)
  • CD4-FITC (Clone OX-38; mouse IgG2A)
  • FoxP3-APC (Goat IgG)
  • Goat IgG-APC Isotype Control
  • Flow Cytometry FoxP3 Staining Buffer (with 1% formaldehyde and 0.09% sodium azide)
  • Flow Cytometry Staining Buffer (with BSA and 0.09% sodium azide)
  • Includes enough reagents to perform 50 assays

 

Other Supplies Required

Other Supplies Required

  • PBS or Hanks’ Balanced Salt Solution (HBSS)

 

Procedure Overview

Cell Staining Protocol with Simultaneous Fixation/Permeabilization

Harvest cells and wash 2 times with PBS or HBSS.

Resuspend cells in Fixation Cytometry Staining Buffer (approximately 1 x 106 cells/100 µL) and transfer to 5 mL flow cytometry tubes.

Cell Staining Protocol Step 1

Add 10 µL CD25 and CD4 fluorochrome-conjugated antibodies.

Incubate the samples at room temperature for 30-45 minutes in the dark.

Cell Staining Protocol Step 2

Wash the cells in Flow Cytometry FoxP3 Staining Buffer.

Cell Staining Protocol Step 3

Add 10 µL FoxP3-APC antibody or goat IgG Isotype Control.

Incubate the samples at room temperature for 1 hour in the dark.

Cell Staining Protocol Step 4

Wash the cells in Flow Cytometry FoxP3 Staining Buffer.

Resuspend the cells in 200-400 μL of Flow Cytometry Staining Buffer.

Cell Staining Protocol Step 5

Analyze the expression of regulatory T cell markers simultaneously by flow cytometry.

Cell Staining Protocol Step 6
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Product Documents for Rat Regulatory T Cell Multi-Color Flow Kit

Certificate of Analysis

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