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Advantages of Fluorescent-labeled Proteins for Detecting Target Molecules

Direct detection. Fluorokines are highly specific and allow target molecules to be directly detected, eliminating the possibility of background staining that may occur by indirect detection using a secondary antibody.

Conjugated to Alexa Fluor® Dyes. Fluorescent-labeled proteins are conjugated to Alexa Fluor® dyes, which offer intense fluorescence and excellent photostability.

Reduced Processing Time. Target detection requires only a single processing step as Fluorokines eliminate the need for staining with a secondary antibody.

High Levels of Bioactivity. Fluorescent-labeled proteins are rigorously tested to ensure that they retain the same high level of activity as the corresponding unlabeled recombinant protein.

High lot-to-lot consistency. Each new lot of a Fluorokine protein is tested side-by-side with previous lots and with a master lot to ensure high lot-to-lot consistency.

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

Fluorokines for Chimeric Antigen Receptor Detection

The ability to evaluate the expression of a chimeric antigen receptor (CAR) following retroviral or lentiviral transduction of a patient’s or donor’s T or natural killer (NK) cells is an important step in the production of CAR-T or CAR-NK cells for cancer immunotherapy. Using fluorescent-labeled proteins, CAR+ cells can be directly stained and detected by flow cytometry, making it easy to evaluate the percentage of transduced cells. Fluorokines are highly specific, reduce processing time, and eliminate background staining that may occur with indirect detection using a secondary antibody.

 

R&D Systems Fluorescent Labeled Proteins for CAR Research

Fluorescent-labeled Proteins for CAR Detection - Products by Molecule

BCMA CD19 CD30/TNFRSF8 CD40/TNFRSF5 CD300e/LMIR6 EGF R
EMMPRIN/CD147 ErbB2/Her2 Glypican 3 Mesothelin MUC-1 OX40
PD-1 Siglec-2/CD22 Siglec-3/CD33 TSLP R    
Fluorokines CAR Expression Illustration

Fluorokines Can Be Used to Directly Assess CAR Expression on CAR-T or CAR-NK Cells

Demonstration of the Utility of a Fluorokine for Evaluating CAR Expression. (A) CAR-T cell therapy is based on the principle that T cells removed from a patient or donor can be genetically engineered to express a specific chimeric antigen receptor (CAR). Once these CAR-T cells are infused back into the patient, the CAR will bind to its specific target antigen on the surface of the patient’s tumor cells, activating the T cells, and allowing them to attack and destroy the tumor cells. (B) The ability to evaluate CAR expression following T cell transduction is an important step in the production of CAR-T cells. T cells expressing the CAR can be directly stained using a Fluorokine (target antigen) and the percentage of CAR-expressing cells can be detected by flow cytometry. (C) CD4+CD8+ T cells were transduced with a hCD19-CAR (left) or not transduced (right) and then cultured for 11 days. Cells were stained with a PE-Cy7-CD4 and Recombinant Human CD19 Fc Chimera Atto 488 Protein (R&D Systems, Catalog # ATJ9269), and detected by flow cytometry.

Fluorokines Are Compatible with Antibody Panels for Multi-Color Flow Cytometry

CAR-T Cell Staining Panel Graphs

Fluorokines can be used with Fluorochrome-conjugated Antibodies to Characterize CD19-CAR-T Cells by Multi-Color Flow Cytometry. 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 (R&D Systems, Catalog # FAB3832N), PerCP-conjugated Mouse Anti-Human CD45 (R&D Systems, Catalog # FAB1430C),PE-conjugated Rabbit Anti-Human CD56/NCAM-1 (R&D Systems, Catalog # FAB24086P), Alexa Fluor® 405-conjugated Mouse Anti-Human CD8 (R&D Systems, Catalog # FAB1509V), PE-Cy7-CD4, and Alexa Fluor® 594-conjugated Mouse Anti-Human CD62L/L-Selectin (R&D Systems, Catalog # 9787T) , along with the Fluorokine, Recombinant Human CD19 Fc Chimera Atto 647N Protein (R&D Systems, Catalog # ATM9269). Cells were initially gated on singlets and live cells.

ATM-193 Consistency Testing Graphs A-C

R&D Systems Fluorokines Undergo Rigorous Testing to Ensure Specificity, Bioactivity, and Lot-to-Lot Consistency

Analysis of the Specificity, Bioactivity, and Lot-to-Lot Consistency of Recombinant Human BCMA/TNFRSF17 Fc Chimera Atto 647N Protein. Fluorescent beads conjugated to an anti-Human BCMA Monoclonal Antibody were stained with (A) Recombinant Human BCMA/TNFRSF17 Fc Chimera Atto 647N Protein (R&D Systems, Catalog # ATM193) or (B) unstained and detected by flow cytometry. (C) Recombinant Human APRIL/TNFSF13 (R&D Systems, Catalog # 5860-AP) was immobilized at 0.1 ug/mL, 100 ul/well, and the indicated concentrations of three independent lots of Recombinant Human BCMA Fc Chimera Atto 647N (R&D Systems, Catalog # ATM193; red, green, orange lines) or unlabeled Recombinant Human BCMA Fc Chimera (R&D Systems, Catalog # 193-BC; blue line) were added. The data demonstrates consistent bioactivity between the fluorescent-labeled and unlabeled proteins and lot-to-lot consistency in the bioactivity of the three different lots of the fluorescent-labeled protein.

Fluorokines for Immune Checkpoint Ligands

Immune checkpoint molecules regulate the activation and functions of different immune cell types and represent some of the most highly investigated targets for cancer immunotherapy. Take advantage of our fluorescent-labeled immune checkpoint ligands to easily identify or sort cells expressing the corresponding immune checkpoint receptors. Fluorescent-labeled ligands allow cells expressing their cognate receptors to be stained in a single step and detected by flow cytometry.

Fluorescent-labeled Immune Checkpoint Ligands - Products by Molecule

B7-1/CD80 B7-2/CD86 B7-H2 CD155/PVR GITR Ligand/TNFSF18 HVEM/TNFRSF14
PD-L1/B7-H1 PD-L2/B7-DC        

Analysis of the Specificity of the Recombinant Human PD-L1/B7-H1 and B7-1/CD80 His-tag Alexa Fluor® 647 Proteins

Alexa Fluor 647 Proteins Graph

Analysis of the Specificity of the Recombinant Human PD-L1/B7-H1 and B7-1/CD80 His-tag Alexa Fluor® 647 Proteins. (A) Streptavidin-coated beads conjugated to Biotinylated Anti-Human PD-L1/B7-H1 Monoclonal Antibody were stained with the indicated concentrations of Recombinant Human PD-L1/B7-H1 His-tag Alexa Fluor® 647 Protein (R&D Systems, Catalog # AFR9049). (B) Streptavidin-coated beads conjugated to Biotinylated Anti-Human B7-1/CD80 Monoclonal Antibody (R&D Systems, Catalog # BAM1402) were stained with the indicated concentrations of Recombinant Human B7-1/CD80 His-tag Alexa Fluor® 647 Protein (R&D Systems, Catalog # AFR9050).

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

Fluorokines for SARS-CoV-2 Spike Proteins

The SARS-CoV-2 Spike protein mediates binding to the host cell ACE-2 receptor, membrane fusion, and viral entry, making it a primary target for potential COVID-19 therapeutics. Our line of fluorescent-labeled SARS-CoV-2 Spike proteins was developed to make it easier for researchers to identify SARS-CoV-2-binding cells, screen for antibodies or small molecules that block the Spike-ACE-2 protein interaction, and investigate SARS-CoV-2-related immune responses. Our catalog includes fluorescent-labeled versions of the SARS-CoV-2 Spike protein full ectodomain and receptor binding domain (RBD), along with fluorescent-labeled SARS-CoV-2 Spike proteins for the alpha, beta, gamma, delta, kappa, and omicron SARS-CoV-2 variants. All fluorescent-labeled Spike proteins are tested to ensure that they detect ACE-2 by flow cytometry.

Fluorescent-labeled SARS-CoV-2 Spike Proteins and ACE-2 - Products by Molecule

ACE-2 SARS-CoV-2 Spike SARS-CoV-2 Spike RBD
Alexa Fluor 488 Proteins Graph

Assessing ACE-2Cells Using Alexa Fluor® 488-Labeled SARS-CoV-2 Spike Protein

Alexa Fluor® Spike Proteins Detect ACE-2 Expression on HEK293 Cells. HEK293 cells transfected with human ACE-2 were stained with (A) 1 ug/mL (100 uL/well) Recombinant SARS-CoV-2 Spike (GCN4-IZ) His-tag Alexa Fluor® 488 Protein (R&D Systems, Catalog # AFG10561) or (B) unstained. Nearly all cells were positive for ACE-2 expression compared to the unstained control.

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

Custom Protein Services

If you don’t see the fluorescent-labeled protein that you need listed on our website, contact our Custom Protein Services team. From scratch protein development to customizing a protein from our catalog, our custom team will work with you to create the protein that meets your experimental needs.

Fluorokine Related Resources

3D image of Avi-Tag protein

Avi-tag Biotinylated Proteins

If you can’t find the fluorescent-labeled protein that you need on our website, explore our wide selection of Avi-tag biotinylated proteins. Avi-tag biotinylated proteins can be used with fluorochrome-labeled streptavidin to detect chimeric antigen receptors, immune checkpoint receptors, or SARS-CoV-2-binding cells.

IO Checkpoints PDL1 Cells

Webinar: How Cancer Circumvents the Immune System

The immune system normally identifies and eliminates cancer cells prior to tumor formation, but cancer cells can induce inhibitory mechanisms that limit anti-tumor immune activity. Watch this webinar to learn more about how cancer cells evade the immune system and what can be done to circumvent or attenuate these mechanisms for therapeutic purposes.

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Flow Cytometry-Validated Antibodies for Identifying Immune Cell Types

The Bio-Techne family brands, R&D Systems and Novus Biologicals, are committed to providing the highest quality antibodies to support your research. Easily identify and characterize different immune cell types using our wide selection of flow cytometry-validated antibodies.

Fluorokine Related Research Areas

Dendritic Immune Cell

Immune Cell Therapy

Harness the power of immune cells to attack specific tumor cell populations. Isolate, expand, engineer, and characterize your cells with our premium reagents, services, and instruments. Our flexible and innovative tools will help to simplify your workflow at every step of the manufacturing process.

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Immune Checkpoint Proteins

Browse our large selection of proteins for studying current and emerging immune checkpoint molecules. In addition to the common targets such as PD-1, PD-L1, and CTLA-4, we offer many lesser-known immune checkpoint proteins that exhibit similar T cell co-inhibitory effects.

SARS-CoV-2 Spike Protein Variants

Proteins for Coronavirus Research

As the COVID-19 pandemic evolves and new SARS-CoV-2 mutant variants are identified, we are striving to provide researchers with the tools that they need to more fully understand this virus. Bio-Techne offers over 100 different SARS-CoV-2 Spike proteins, including multiple different versions of the Alpha, Beta, Gamma, Delta, Omicron, and IHU Spike protein variants.