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Découvrez notre série de protéines de haute qualité couvrant des indicateurs diagnostiques complets afin de faciliter la recherche diagnostique in vitro des maladies neurologiques.
Les fibrilles préformées constituent un modèle préclinique inestimable pour explorer la pathogenèse des maladies neurologiques par l'agrégation de protéines mal repliées. Découvrez nos PFF pré-ensemencées à utiliser dans le cadre de vos recherches.
Les facteurs neuronaux sont une classe de molécules protéiques dotées d'une activité neurotrophique qui peut favoriser la survie et la régénération des cellules nerveuses. Découvrez notre série de facteurs neuronaux recombinants destinés à favoriser la culture et la différenciation des cellules nerveuses.
En partenariat avec Diagnostic Biochips, nous proposons désormais des solutions pour les enregistrements électrophysiologiques in vivo, notamment des électrodes multicanaux de haute qualité et d'autres produits destinés à faciliter l'analyse efficace et de haute qualité de la structure et de la fonction des circuits neuronaux.
Neural antibodies can specifically label and recognize molecules on nerve cells, enabling a more comprehensive understanding and study of the biological properties, functions, and mechanisms of nerve cells in neurodegenerative diseases.
Les marqueurs protéiques sont un élément essentiel de la recherche biologique et du développement de médicaments. Que ce soit pour l'électrophorèse des protéines ou le western blot, nos marqueurs protéiques pré-colorés vous aident à déterminer rapidement le poids moléculaire de la protéine cible ou à évaluer l'efficacité du transfert.
La streptavidine est une protéine tétramérique offrant 4 sites de liaison à la biotine de haute affinité. Nous proposons une large gamme de produits préconjugués à la streptavidine pour soutenir votre recherche ainsi que des protéines biotinylées.
L'utilisation de l'anticorps de contrôle isotype correspondant pour détecter la liaison non spécifique permet de réduire la production de résultats faussement positifs et d'évaluer avec précision les facteurs d'influence possibles au cours du processus de développement du médicament. Découvrez nos contrôles d'isotypes pour votre recherche.
Vous souhaitez développer votre propre test de chimiluminescence ? Explorez nos produits pour assembler votre propre MPCLIA en fonction de vos besoins !
L'efficacité d'un anticorps thérapeutique dépend du fragment Fab et de son activité de liaison à l'antigène cible, mais aussi du fragment Fc et de son interaction avec les principaux récepteurs Fc. Par conséquent, les candidats doivent être testés sur un ensemble de récepteurs durant l’ingénierie de l’anticorps. Explorez notre collection complète de protéines recombinantes du récepteur Fc !
Nous nous sommes engagés à accélérer la recherche, le développement, l'approbation et la commercialisation de vaccins contre les maladies infectieuses. Avec cette idée, ViruStop est spécialement conçu pour la recherche sur les virus !
Des anticorps spécifiques à l'antigène ou neutralisants sont disponibles pour votre recherche ! Que vous cherchiez à réaliser des tests de neutralisation ou à évaluer les titres d'anticorps, explorez notre catalogue d'anticorps !
Au lieu de développer vos propres tests, découvrez nos kits et tests conçus pour quantifier rapidement la présence d'antigènes ou le titre d'anticorps !
Evaluating CAR expression is an essential step in the production of CAR-T cells. This is often done by flow cytometry,using protein L, anti-Fab antibodies, anti-idiotypic antibody or target antigens as detection antibodies. Among these common choices, target antigens are widely considered to be the best option, because it offers high specificity and minimal background staining.
Click to download the Guidance for CAR detection
Reagents
Mechanism
Pros
Cons
Target Antigens
Specifically bind to the antigen-binding domains of CARs.
High specificity
Minimal background staining
High cost
Each unique CAR has to be stained with corresponding antigens
Protein L
Binds to the kappa light chain of immunoglobulin.
Universal and low cost
High background staining.
Cannot detect the anti-lambda light chain CAR
Anti-Fab antibody
Binds to the Fab portion of immunoglobulin.
Universal and low cost
High background staining
Anti-idiotypic Antibody
Specifically bind to the antigen-binding domains of CARs.
High specificity;
High sensitivity
Minimal background staining.
Each unique CAR has to be stained with corresponding Anti- idiotypic Antibody.
Most need to be customized and customization is very time-consuming
Case No.1 Evaluation of anti-BCMA CAR expression using biotinylated BCMA
Reagents:Biotinylated human BCMA protein, Fc & Avi Tag (ACROBiosystems, Cat. No. BC7-H82F0); PE Streptavidin (Biolegend, Cat. No. 405204).
Samples:Jurkat cells expressing BCMA-CAR ; Primary T cells expressing BCMA-CAR
Brief Protocol:
1.Transduce human primary T cells or Jurkat T cells with BCMA-CAR lentivirus.2.Collect the CAR transduced human T cells and un-transduced T cells.3.Count the cells and aliquot 2 x105 cells for staining into each tube.4.Wash the cells once with a FACS buffer.5.Dilute the biotinylated human BCMA protein (ACROBiosystems, Cat. No. BC7-H82F0) with working stock to 3µg/ml. Add 100µl of Biotinylated BCMA to each tube and incubate for 60 minutes in ice (protect from light).6.Wash the cells with FACS buffer once and stain with 100µl of 1µg/ml PE conjugated streptavidin for 30 minutes in ice (protect from light).7.Wash the cells twice with FACS buffer.8.Resuspend the cells in 200µl FACS buffer.9.Analyze the results using Attune NxT flow cytometer and FlowJo software.
Results:The data show that BCMA-CAR expression can be successfully quantified using Biotinylated BCMA from ACROBiosystems (Cat. No. BC7-H82F0) both in jurkat cell line (Fig.1) and primary T cells (Fig.2). Control cells lacking BCMA-CAR expression showed no positive staining (Fig. 3) with PE- Streptavidin conjugate indicating that the staining using biotinylated BCMA is specific.
Fig. 1. BCMA-CAR expression in Jurkat cells
Fig. 2. BCMA-CAR expression in Primary T cells
Fig. 3 Control cells without BCMA-CAR expression.
Data are kindly provided by R&D team, Theragent Inc
Reagents:Biotinylated human BCMA protein, Fc & Avi Tag (ACROBiosystems, Cat. No. BC7-H82F0);PE Streptavidin (Biolegend, Cat. No. 405204).
Samples:Anti-BCMA CAR-transduced human primary T-cells.
Brief Protocol:
1.Transduce human primary T-cells with a lentiviral vector to express anti-BCMA CAR;2.Three days post-transduction, cells were stained with a biotinylated human BCMA protein (ACROBiosystems, Cat. No. BC7-H82F0);3.After washing, secondary labeling was performed with PE Streptavidin;4.The cells were analyzed using BD FACSCaliburTM flow cytometer, and the data was analyzed with FCS Express 6 Plus software.
Results:The data showed that the expression of anti-BCMA CARs on transduced T cell surface from donor 1 and donor 2 were 52.72% and 73.49%, respectively.
Human T cells were transfected with anti-BCMA CAR and cultured for 3 days. Three days post-transfection, 1x106 cells were first incubated with 50ul biotinylated human BCMA protein (Cat. No. BC7-H82F0, 8ug/ml ), washed and then stained with PE Streptavidin and analyzed by flow cytometry.
Data are kindly provided by PREGENE Biopharma
Case No.2 Evaluation of anti-CD19 CAR expression using FITC-labeled CD19 protein
Reagents:FITC-labeled Human CD19 (20-291) Protein (ACROBiosystems, Cat. No. CD9-HF2H2).
Samples:Anti-CD19 CAR-293 cells.
Brief Protocol:
1.Culture anti-CD19 CAR-293 cells in DMEM medium with 10% FBS in the CO2 incubator (at 37°C, 5% CO2). 2.Harvest the cells and wash the cells once by wash buffer. 3.Count the cells number and the viability, aliquot up 2e5 live cells (Anti-CD19-scFv positive cell is 98%) into each tube. (Note: the cell viability must be ≥ 95%.) 4.Add 100 µl, 10 µg/ml of FITC-labeled Human CD19 (20-291) Protein (ACROBiosystems, Cat. No. CD9-HF2H2) or FITC-labeled Protein control into each tube, incubating at 4℃ for 1 hour. 5.Wash the cells 3 times by wash buffer and resuspend the cells in 200 µl PBS per sample. 6.Transfer the cells into flow tube and detect by Flow cytometry. 7.Analyze result using FACS Celesta software and FCS Express 6 Flow software.
Results:The data showed that the expression level of anti-CD19 scFv on the surface of anti-CD19 CAR-293 cells was 95.37%.
293 cells were transfected with anti-CD19-scFv and RFP tag. 2e5 of the cells were stained with B. FITC-Labeled Human CD19 (20-291) (Cat. No. CD9-HF2H2, 10 µg/ml) and C. FITC-labeled protein control. A. Non-transfected 293 cells and C. FITC-labeled protein control were used as negative control. RFP was used to evaluate CAR (anti-CD19-scFv) expression and FITC was used to evaluate the binding activity of FITC-labeled Human CD19 (20-291) (Cat. No. CD9-HF2H2).
Case No.3 Evaluation of anti-CD19 CAR expression using FITC-labeled anti-FMC63 scFv antibody
1.Culture Anti-CD19 CAR-293 cells in DMEM medium with 10% FBS in the CO2 incubator (at 37 ℃, 5% CO2). 2.Harvest the cells and wash the cells once by FACS buffer. 3.Count the cells number and the viability, aliquot up 2e5 live cells into each tube. (Note: the cell viability must ≥ 95%.) 4.Dilute FITC-Labeled Monoclonal Anti-FMC63 scFv Antibody, Mouse IgG1 (ACROBiosystems, Cat. No. FM3-FY45) in FACS buffer to get the working solution just before the assay, and then add 100 μL of the working solution into the tube with cell pellet. Mix well and incubate at 4℃ for 60 minutes. 5.Wash the cells 3 times by FACS buffer and resuspend the cell pellet in 200 μL PBS per sample. 6.Transfer the cell suspension into flow tube and detect the cells by Flow cytometry. 7. Analyze the result data using FCS Express 7 Plus and GraphPad Prism 5 software.
Results:The data showed that the expression level of anti-CD19 scFv on the surface of anti-CD19 CAR-293 cells was 100%.
2e5 of Anti-CD19 CAR-293 cells were stained with 100 μL of 1:50 dilution (2 μL stock solution in 100 μL FACS buffer) FITC-Labeled Monoclonal Anti-FMC63 scFv Antibody, Mouse IgG1 (Cat. No. FM3-FY45) and isotype control respectively. FITC signal was used to evaluate the binding activity .
Case No.4 Evaluation of anti-MSLN CAR expression using PE-labeled MSLN protein
Reagents:PE-labeled Human Mesothelin / MSLN (296-580) Protein (ACROBiosystems, Cat. No. MSN-HP2H5).
Samples:Anti-MSLN CAR-293 cells
Brief Protocol:
1.Culture anti-MSLN CAR-293 cells in DMEM medium with 10% FBS in the CO2 incubator (at 37 ℃, 5% CO2). 2.Harvest the cells and wash the cells once by wash buffer. 3.Count the cells number and the viability, aliquot up 1e6 live cells into each tube. 4.Add 100 µl of diluted PE-labeled Human Mesothelin (296-580) Protein (Cat. No. MSN-HP2H5) (prepared in dilution buffer at 1:50 dilution) into each tube, incubating at 4℃ for 1 hour. 5.Wash the cells 3 times by wash buffer and resuspend the cells in 200 µl PBS per sample. 6.Transfer the cells into flow tube and detect by Flow cytometry. 7.Analyze result using FACS Celesta software and FCS Express 6 Flow software.
Results:The data showed that the expression level of anti-MSLN scFv on the surface of anti-MSLN CAR-293 cells was 100 %.
1e6 of the anti-MSLN CAR-293 cells were stained with 100 μL of 1:50 dilution (2 μL stock solution in 100 μL FACS buffer) of PE-Labeled Human Mesothelin (296-580) Protein, His Tag (Cat. No. MSN-HP2H5) and negative control protein respectively, PE signal was used to evaluate the binding activity.
Case No.5 Evaluation of the detection specificity of CD19, Fc Tag
Reagents:Human CD19 Protein, Fc Tag (ACROBiosystems, Cat. No. CD9-H5259);Human CD19 Protein ,Fc Tag (Company N);Human PD-L1 Protein, Fc Tag (ACROBiosystems, Cat. No. PD1-H5258), as negative control;FITC anti-human IgG Fc antibody (Biolegend, Cat. No. 409310).
1.Cells were stained for CAR expression using ACRO’s human CD19 protein, Fc Tag or Company N’s human CD19 protein, Fc Tag;2.After washing, secondary labeling was performed with FITC anti-human IgG Fc antibody;3.The cells were analyzed using NovoCyteTM flow cytometer and the data analyzed by FCS Express 6 Plus and GraphPad Prism 5 software.
Results:The data shows that Company N’s human CD19-Fc fusion protein exhibits strong non-specific binding to 293 and Jurkat cells.
Binding specificity analysis of ACROBiosystems hCD19(C-Fc tag) protein
FACS analysis of human CD19 protein, Fc Tag (ACROBiosystems, Cat. No. CD9-H5259) binding to A. R1013-C6 cells, B. Expi 293 cells, C. Jurkat E6.1 cells. Cells were first stained with human CD19 protein, Fc Tag (ACROBiosystems, Cat. No. CD9-H5259) followed by FITC anti-human IgG Fc antibody, and then analyzed using NovoCyteTM Flow Cytometer. The data were analyzed with FCS Express 6Plus and GraphPad Prism 5 software.
Binding specificity analysis of Company N hCD19(C-Fc tag) protein
FACS analysis of human CD19 protein, Fc Tag (Company N) binding to A. R1013-C6 cells, B. Expi 293 cells, C. Jurkat E6.1 cells. Cells were first stained with human CD19 protein, Fc Tag (Company N) followed by FITC anti-human IgG Fc antibody, and then analyzed using NovoCyteTM Flow Cytometer. The data were analyzed with FCS Express 6Plus and GraphPad Prism 5 software.