Skip to main content

Easily Identify and Phenotype CAR-T Cells by Flow Cytometry

After transduction of T cells or NK cells with the chimeric antigen receptor (CAR), it is important to assess the degree of transduction (purity) as well as understand the phenotype of transduced cells (potency). Bio-Techne’s fluorescent conjugated antibodies and Fluorokine™ proteins allow you to assess this in one simple staining step.

 

CAR Cell Panel

Marker Clone Fluorochrome Catalog Number
CD14

134620

Alexa Fluor® 700

FAB3832N

CD45 2D1 PerCP

FAB1430C

CD56/NCAM-1 2524C PE

FAB24086P

CD8 37006

Alexa Fluor® 405

FAB1509V

CD4

-

PE-Cy7TM

Available from Novus Biologicals

CD62L/L-Selectin

4G8

Alexa Fluor® 594

FAB9787T

Human CD19 Fc Chimera

-

Atto 647

ATM9269

CAR-T Cell Staining Panel Graphs

FluorokinesTM Can be Used with Fluorochrome-conjugated Antibodies to Characterize CD19-CAR-T Cells by Multi-Color Flow Cytometry. Human PBMC CD4+CD8+ T cells transduced with a hCD19-CAR were stained with the following panel of monoclonal antibodies: Alexa Fluor® 700-conjugated Mouse Anti-Human CD14 (Catalog # FAB3832N), PerCP-conjugated Mouse Anti-Human CD45 (Catalog # FAB1430C), PE-conjugated Rabbit Anti-Human CD56/NCAM-1 (Catalog # FAB24086P), Alexa Fluor® 405-conjugated Mouse Anti-Human CD8 (Catalog # FAB1509V), PE-Cy7-CD4, and Alexa Fluor® 594-conjugated Mouse Anti-Human CD62L/L-Selectin (Catalog # FAB9787T), along with the Fluorokine, Recombinant Human CD19 Fc Chimera Atto 647N Protein (Catalog # ATM9269) and detected by flow cytometry. Cells were initially gated on singlets and live cells. 

Alexa Fluor® is a registered trademark of Molecular Probes, Inc., Eugene, OR.

Fluorokine App Note Cover

Evaluating CAR Expression on CAR-T Cells Using FluorokinesTM

One of the current challenges in the field of CAR-T cell therapy is the lack of a simple assay to easily detect and quantify CAR expression following T cell transduction. Read this application note to learn about the advantages of using R&D SystemsTM Fluorokines fluorescent-labeled proteins to directly stain and detect CAR+ cells by flow cytometry.

BioInsights Webinar Foreground

The CAR ToolKit: Progress and Challenges in CAR-T/NK Cell R&D and Mfg

With numerous clinical trials underway and multiple CAR-T cell products now approved, solving the current obstacles when manufacturing CAR-T/NK cell products continues to be a pressing issue for the cellular immunotherapy field. Watch this webinar, hosted by BioInsights, to learn about the current challenges, latest progress, and innovation facing CAR-T and CAR-NK cell development.

Rainbow-colored mesh design over dark blue background.

Flow Cytometry Panel Builder Tool

 

The Bio-Techne Panel Builder tool is designed to prevent selection of incompatible fluorochromes by blocking channels with excessive spectral overlap. Save time and reduce costly mistakes by quickly finding compatible reagents for multicolor flow cytometry experiments using the Panel Builder Tool.

Fluorescent Conjugated Proteins for CAR Detection

The ability to evaluate the expression of a chimeric antigen receptor (CAR) following T cell or natural killer (NK) cell transduction is a critical step in the production of CAR-T or CAR-NK cells. FluorokineTM proteins are directly conjugated with fluorescent molecules like Alexa Fluor® 647, Alexa Fluor® 488, Atto 488, and Atto 647. Fluorokines are highly specific, reduce processing time, and eliminate background staining that may occur by indirect detection of the CAR using an epitope-tagged target antigen and a fluorophore-labeled secondary antibody.

 

Fluorescent-Labeled Proteins for CAR-T or CAR-NK Cell Detection

BCMA CD19 CD30/TNFRSF8 CD40/TNFRSF5
CD300e/LMIR6 DLL3 EGF R EMMPRIN/CD147
EpCAM/TROP1 ErbB2/Her2 Fc gamma RIIA/CD32a (R167) Fc gamma RIIIA/CD16a
Glypican 3 IL-13 R alpha2 Mesothelin MUC-1
OX40 PD-1 Siglec-2/CD22 Siglec-3/CD33
TRAIL R2 TSLP R VEGF R2/KDR  

Custom Fluorokine Solutions

Unable to find the fluorescent-labeled protein that you need? Contact our trusted Custom Protein Services team can work with you to create a customized protein solution to meet your specific research needs.

Flow Cytometry Validated Antibodies

At Bio-Techne, we know the importance of quality. Our flow cytometry antibodies are backed by our 100% Guarantee so you can have confidence in your purchase.

Antibodies to Phenotype T Cells

Memory T Cell Markers

T Cell Exhaustion Markers

CD3

PD-1

CD4

LAG-3

CD8

TIM-3

CD45RA*

TIGIT

CD45RO

 

CCR7

T Cell Activation Markers

L-Selectin/CD62L

HLA-DR

CD27/TNFSF7

CD38

CD28

CD69

Fas/TNFRSF6/CD95

CD25/IL-2R alpha

The Fastest Way to a Single Cell

Hana and Pala from Namocell, now a Bio-Techne brand help you sort and dispense cells into 96-well or 384-well plates or 2mL tubes on your own bench. Three patented sorting modes:

  • Single cell sorting mode dispenses single cells one-cell-per-well.
  • Enrichment mode isolates rare cells from extremely large samples.
  • Bulk sorting mode sorts tens of thousands cells in bulk.

Fluorescent Dyes

7 Aminoactinomycin D Solution Flow Cytometry NBP2 29446

Cell Viability Dyes

dapi 5748 spectra

DNA Dyes

DRAQ9 TM Immunocytochemistry Immunofluorescence NBP2 81128

Non-nuclear Dyes

Quality Control and Standardization Beads

Set yourself up for successful flow cytometry experiments by using our Calibration, Compensation, Cell Counting, and Equivalent Reference Fluorochromes (ERF) beads.

Complete Your CAR T Cell or CAR NK Cell Workflow

Bio-Techne is here to be your partner through every stage of the CAR T Cell and CAR NK Cell Therapy workflow, from cell isolation through in vivo monitoring.

Learn more about T Cell Manufacturing and NK Cell Manufacturing with Bio-Techne.

ExCellerate Cell Culture

Immune Cell Culture and Expansion

We offer a wide selection of high-quality cell culture media, fetal bovine serum, and recombinant proteins for improved survival and expansion of different immune cell types.

Cell Culture Development GMP Proteins

GMP Proteins for Cell and Gene Therapy

Discover the latest GMP and animal-free RUO proteins available from our state-of-the-art animal-free GMP manufacturing facility located in Saint Paul, Minnesota.

ScaleReady G REX

Scale Up with ScaleReady

Advance your cultures with an innovative cell expansion platform that alleviates scale-up challenges. Our partners at ScaleReady offer G-Rex® 6M well plates and closed-system bioreactors that allow the same protocols to be used from early research through commercial scale up, dramatically simplifying transition to GMP manufacturing.

gowned cleanroom operator carrying cell culture vessel

Transition to Animal-Free and GMP

We understand the need for adopting increasingly well-defined raw materials. Regardless of where your program is in this trajectory, you can rely on us to provide multiple grades of key materials – with flexibility and customization to fit your process.