Cd4 rpa t4

Manufactured by BD

Sourced in United States

The CD4 (RPA-T4) is a lab equipment product designed for the detection and quantification of CD4+ T cells. It functions as a biomarker to monitor immune system status. The product utilizes flow cytometry technology to analyze cell samples.

Automatically generated - may contain errors

Lab products found in correlation

Cd3 sk7, by BD (5 mentions) Cd8 sk1, by BD (5 mentions) Brefeldin a, by Merck Group (3 mentions) Ionomycin, by Merck Group (3 mentions) Live dead dye, by Thermo Fisher Scientific (2 mentions) Ifn γ b27, by BD (2 mentions) Il 2 mq1 17h12, by BioLegend (2 mentions) Cxcr5 rf8b2, by BD (2 mentions) Cd25 m a251, by BD (2 mentions) Live dead blue, by Thermo Fisher Scientific (2 mentions) Cd45ro uchl1, by Beckman Coulter (2 mentions) Il 4 8d4 8, by BD (2 mentions) Il 5 jes1 39d10, by BD (2 mentions) Il 13 jes10 5a2, by BD (2 mentions) Cytofix cytoperm, by BD (2 mentions) Cd27 o323, by BioLegend (2 mentions) Cd19 sj25c1, by BD (2 mentions) Cd20 2h7, by BD (2 mentions) Cd38 hb7, by BD (2 mentions) Facsymphony a5, by BD (2 mentions)

16 protocols using cd4 rpa t4

Multiparameter Immune Cell Profiling

Check if the same lab product or an alternative is used in the 5 most similar protocols

Live/Dead dye (Invitrogen), CD4 (RPA-T4, BD Biosciences), CD3 (UCHT1), CD8 (5K1), Vα7.2 (3C10), and CD161 (191B8) [all Biolegend].

Phetsouphanh C., Phalora P., Hackstein C.P., Thornhill J., Munier C.M., Meyerowitz J., Murray L., VanVuuren C., Goedhals D., Drexhage L., Russell R.A., Sattentau Q.J., Mak J.Y., Fairlie D.P., Fidler S., Kelleher A.D., Frater J, & Klenerman P. (2021). Human MAIT cells respond to and suppress HIV-1. eLife, 10, e50324.

+ Open protocol

+ Expand

Flow Cytometry Analysis of Regulatory T Cells

Check if the same lab product or an alternative is used in the 5 most similar protocols

The following conjugated mouse antibodies were used for flow cytometry analysis (clone [fluorophore]): anti-CD3 (HIT3a [FITC]), CD4 (RPA-T4 [PE and PerCP]), CD45RA (HI100 [APC]) CD25 (M-A251 [APC] or 2A3 [FITC]), CD69 (FN50 [PE]), CD127 (HIL-R7-M21 [FITC]), TNFR2 (hTNFR-M1 [PE]) and CTLA-4 (BNI3 [PE]) (BD Pharmingen, San Jose-CA, USA). The anti-GITR conjugated antibody (621 [APC]) was obtained from BioLegend. The anti-LAP conjugated antibody (27232 [PE]) was obtained from R&D Systems (Minneapolis, MN, USA) and PE conjugated anti-human FOXP3 (PCH101) was obtained from eBiosciences (San Diego, CA, USA). For FOXP3 intracellular staining, the cells were fixed and permeabilized using a Human FOXP3 Human Buffer Set kit (BD Pharmingen, San Jose-CA, USA). The reagents and cytokines used included: RPMI 1640 medium and all-trans retinoic acid from Sigma-Aldrich (St. Louis, MO, USA), recombinant human TGF-β1 and IL-2 from Peprotech (Rocky Hill, NJ, USA), anti-CD2/CD3/CD28 beads (T Cell Activation/Expansion Kit) and carboxyfluoresceinsuccinimidyl ester (CFSE) from Invitrogen (Carlsbad, CA, USA).

Schiavinato J.L., Haddad R., Saldanha-Araujo F., Baiochi J., Araujo A.G., Santos Scheucher P., Covas D.T., Zago M.A, & Panepucci R.A. (2017). TGF-beta/atRA-induced Tregs express a selected set of microRNAs involved in the repression of transcripts related to Th17 differentiation. Scientific Reports, 7, 3627.

+ Open protocol

+ Expand

Characterization of CD1d-Reactive T Cells

Check if the same lab product or an alternative is used in the 5 most similar protocols

Enriched CD3⁺ CD1d–α-GalCer tetramer⁺ cells were stained with Vδ1 (clone A13 supernatant, which can bind to Vδ1 when incorporated in hybrid Vδ1-Jα-Cα TCR chains, was produced in L. Moretta’s laboratory), anti–mouse IgG (clone Poly 4053; BioLegend), 5% normal mouse serum, and then with antibodies specific for αβTCR (clone T10B9.1A-31; BD), CD3ε (clone UCHT1; BD), CD8α (SK1; BD), CD4 (RPA-T4; BD), Vβ11 (C21; Beckman Coulter), CD69 (FN50; BD), γδTCR (11F2; BD), and CD161 (191B8; Miltenyi Biotec). Cells were costained with 7-aminoactinomycin D viability dye (Sigma-Aldrich) and with human CD1d tetramers (produced in-house), as previously described (Uldrich et al., 2013 (link)). TCR-Vβ repertoire analysis was performed using a TCR-Vβ repertoire kit (Beckman Coulter). Cells were analyzed using an LSR Fortessa (BD), and data were analyzed using FlowJo software (Tree Star Inc.).

Pellicci D.G., Uldrich A.P., Le Nours J., Ross F., Chabrol E., Eckle S.B., de Boer R., Lim R.T., McPherson K., Besra G., Howell A.R., Moretta L., McCluskey J., Heemskerk M.H., Gras S., Rossjohn J, & Godfrey D.I. (2014). The molecular bases of δ/αβ T cell–mediated antigen recognition. The Journal of Experimental Medicine, 211(13), 2599-2615.

+ Open protocol

+ Expand

Activation and Phenotyping of T Cells

Check if the same lab product or an alternative is used in the 5 most similar protocols

Patients’ and healthy control (HC) PBMCs were purified by Ficoll density gradient centrifugation, washed with RPMI 1640 plus penicillin, streptomycin, and L-glutamine along with 10% FBS (R10) and filtered through a 40 µM strainer. Cells were resuspended to a concentration of 1 × 10⁶ cells/mL and aliquoted (1 mL/well) into 48-well plates. PMA (final concentration 20 ng/mL), ionomycin (1 µM) and brefeldin A (5 µg/mL, Sigma-Aldrich) were added prior to incubation at 37°C for 5–6 hours. After incubation, cells were washed with FACS buffer and stained with LIVE/DEAD Blue (Life Technologies). Intracellular staining was performed using BD CytoFix/CytoPerm (BD Biosciences) reagents according to the manufacturer’s instructions using the following antibodies: CD3 (SK7, BD Biosciences), CD4 (RPA-T4, BD Biosciences), CD8 (SK1, BD Biosciences), CD45RO (UCHL1, Beckman Coulter), CXCR5 (RF8B2, BD Biosciences), CD25 (M-A251, BD Biosciences), IL-2 (MQ1–17H12, BioLegend), IL-4 (8D4–8, BD Biosciences), IL-5 (JES1–39D10, BD Biosciences), IL-13 (JES10-5A2, BD Biosciences), IFNγ (B27, BD Biosciences), IL-17A (eBio64DEC17, eBioscience). Cells were gated on viable CD3⁺ CD4⁺ CD45RO⁺ cells. Ex-vivo Treg staining was performed using anti-CD3 (UCHT1), CD4 (OKT4), CD25 (M-A251),, CD45RO (UCHL1), CD127 (HIL-7R–M21),, GATA3 (L50–823) and FOXP3 (236A/E7) antibodies as described ^{33 (link)}.

Ma C.A., Stinson J.R., Zhang Y., Abbott J.K., Weinreich M.A., Hauk P.J., Reynolds P.R., Lyons J.J., Nelson C.G., Ruffo E., Dorjbal B., Glauzy S., Yamakawa N., Arjunaraja S., Voss K., Stoddard J., Niemela J., Zhang Y., Rosenzweig S.D., McElwee J.J., DiMaggio T., Matthews H.F., Jones N., Stone K.D., Palma A., Oleastro M., Prieto E., Bernasconi A.R., Dubra G., Danielian S., Zaiat J., Marti M.A., Kim B., Cooper M.A., Romberg N.D., Meffre E., Gelfand E.W., Snow A.L, & Milner J.D. (2017). Germline hypomorphic CARD11 mutations in severe atopic disease. Nature genetics, 49(8), 1192-1201.

+ Open protocol

+ Expand

Proliferation and STAT1 Activation Assay

Check if the same lab product or an alternative is used in the 5 most similar protocols

PBMCs were incubated with CellTrace violet (1 μM, Cell Proliferation Kit, Thermo), stimulated with anti-IgM (10 μg/ml, Jackson Lab; 109-006-129), CD40L (100 ng/ml, Enzo; ALX522-110-C010), IL-4 (10 ng/ml, Peprotech; 200-04), IL-21 (100 ng/ml, Peprotech; 200-21), CpG (1μM, Enzo; ALX746-006-C100), BAFF (100 ng/ml, Enzo ALX-522-025-C010), anti-CD3 (1 μg/ml, eBioscience; 16-0037-85), anti-CD28 (1 μg/ml, eBioscience; 16-0289-85) and PHA (1 μg/ml, Sigma; L9017). Seventy-two hours later the cells were stained with surface markers and analyzed by flow cytometer. Staining was performed with following antibodies from BD: CD3 (UCHT1), CD4 (RPAT4), CD8 (RPAT8), CD19 (HIB19) and CD80 (L307.4). For p-STAT1 detection, staining was performed using following antibodies from BD: CD4 (RPAT4), CD8 (SK1) and p-STAT1 (4a), and from Biolegend: CD3 (UCHT1), CD14 (M5E2) and CD45RA (HI100).

Oda H., Beck D.B., Kuehn H.S., Sampaio Moura N., Hoffmann P., Ibarra M., Stoddard J., Tsai W.L., Gutierrez-Cruz G., Gadina M., Rosenzweig S.D., Kastner D.L., Notarangelo L.D, & Aksentijevich I. (2019). Second Case of HOIP Deficiency Expands Clinical Features and Defines Inflammatory Transcriptome Regulated by LUBAC. Frontiers in Immunology, 10, 479.

+ Open protocol

+ Expand

Multicolor Flow Cytometry Panel

Check if the same lab product or an alternative is used in the 5 most similar protocols

Live/Dead dye (Invitrogen), CD4 (RPA-T4, BD Biosciences), CD3 (UCHT1), CD8 (5K1) and CD57 (HNK-1)[all Biolegend]. Added to all flow cytometry-staining panels.

Phetsouphanh C., Aldridge D., Marchi E., Munier C.M., Meyerowitz J., Murray L., Van Vuuren C., Goedhals D., Fidler S., Kelleher A., Klenerman P, & Frater J. (2019). Maintenance of Functional CD57+ Cytolytic CD4+ T Cells in HIV+ Elite Controllers. Frontiers in Immunology, 10, 1844.

+ Open protocol

+ Expand

Immunofluorescence Analysis of CD4+ T Cells

Check if the same lab product or an alternative is used in the 5 most similar protocols

For immunofluorescence analysis, freshly isolated CD4⁺ T cells were attached to poly-L-lysine coated glass slides by 30 minutes of incubation at room temperature and stained with CD4 (RPA-T4) (BD Biosciences) and BMPRIA (R&D Systems) antibodies followed by Alexa Fluor 594 or Alexa Fluor 488-conjugated multiadsorbed F(ab’)2 fragments of donkey anti-mouse and donkey anti-goat IgG. For intracellular staining of BMPRIA, cells were fixed with Cytofix/Cytoperm solution (BD Biosciences) before CD4 staining and stained for BMPRIA using Perm/Wash buffer (BD Biosciences). Finally, cells were stained with Hoechst 33342 (Invitrogen, Life Technologies) for cell nucleus visualization and mounted with Prolong Gold (Invitrogen, Life Technologies). Slides were examined on a FluoView 1200 Confocal Microscope (Olympus) from the Centro de Citometría y Microscopía de Fluorescencia, Complutense University of Madrid.

Martínez V.G., Sacedón R., Hidalgo L., Valencia J., Fernández-Sevilla L.M., Hernández-López C., Vicente A, & Varas A. (2015). The BMP Pathway Participates in Human Naive CD4+ T Cell Activation and Homeostasis. PLoS ONE, 10(6), e0131453.

+ Open protocol

+ Expand

Multicolor Flow Cytometry of PBMCs and Lung Lymphocytes

Check if the same lab product or an alternative is used in the 5 most similar protocols

Monoclonal antibodies used to stain human PBMCs were from BioLegend CD27 (O323), CD8α (RPA-T8), CD3 (OKT3), IFN-γ (4S.B3), TNFα (MAb11), and BD Biosciences CD4 (RPA-T4). Monoclonal antibodies used to stain lung-derived murine lymphocytes were BioLegend CD3 (17A2), CD8α (53–6.7), CD4 (RM4–5); eBioscience CD11b (M1/70), CD44 (IM7); and BD Biosciences IFN-γ (XMG1.2). Samples were run on a BD Biosciences LSRII flow cytometer and analyzed with FlowJo software.

McMaster S.R., Gabbard J.D., Koutsonanos D.G., Compans R.W., Tripp R.A., Tompkins S.M, & Kohlmeier J.E. (2015). Memory T Cells Generated by Prior Exposure to Influenza Cross React with the Novel H7N9 Influenza Virus and Confer Protective Heterosubtypic Immunity. PLoS ONE, 10(2), e0115725.

+ Open protocol

+ Expand

Phenotyping Antigen-Specific B cells

Check if the same lab product or an alternative is used in the 5 most similar protocols

Freshly isolated peripheral blood mononuclear cells were stained first for viability with Live/dead Yellow (ThermoFisher) and then for markers with the following monoclonal antibodies: IgA (IS11–8E10, Miltenyi), IgD (IA6–2, BD), IgG (G18–145, BD), IgM (MHM-88, Biolegend), CD3 (SK7, BD), CD4 (RPA-T4, BD), CD8 (SK1, BD), CD14 (61D3, eBioscience), CD16 (CB16, eBioscience), CD19 (SJ25C1, BD), CD20 (2H7, BD), CD27 (O323, BioLegend or M-T271, BD), CD38 (HB7, BD), and CD71 (CY1G4, BioLegend. Antigen-specific B cells were detected by staining with RBD conjugated to Alexa Fluor 488 (Protein Labeling Kit, ThermoFisher). RBD was conjugated according to manufacturer’s instructions, with the following changes: protein was labeled at a concentration of 1mg/mL, and incubated for 30 minutes without the addition of bicarbonate. After staining, PBMCs were washed and then fixed for 30 minutes using 2% paraformaldehyde (ThermoFisher). Data were acquired on a BD FACSymphony A5 and analyzed using FlowJo 10.7.1 (BD).

Mantus G., Nyhoff L.E., Kauffman R.C., Edara V.V., Lai L., Floyd K., Shi P.Y., Menachery V.D., Edupuganti S., Scherer E.M., Kay A., McNair N., Anderson E.J., Rouphael N., Ahmed R., Suthar M.S, & Wrammert J. (2021). Evaluation of Cellular and Serological Responses to Acute SARS-CoV-2 Infection Demonstrate the Functional Importance of the Receptor Binding Domain. Journal of immunology (Baltimore, Md. : 1950), 206(11), 2605-2613.

+ Open protocol

+ Expand

Multiparametric Flow Cytometry of T Cell Responses

Check if the same lab product or an alternative is used in the 5 most similar protocols

Tumour cells or T cells suspended in FACS staining buffer were stained with fluorochrome-conjugated antibodies against combinations of the following surface human antigens: CD4 (RPA-T4, BD, Franklin Lakes, NJ), CD8 (SK1, BD), CD3e (SK7, BD), HLA-A*02/MART-1:27–35 tetramer (DMF5, Beckman Coulter Immunotech, Monrovia, CA); NY-ESO-1 tetramer (NIH Tetramer Core Facility); PD1-L1 (MIH1, eBiosciences, San Diego, CA); PD1-L2 (24F.10C12, Biolegend, San Diego, CA); IFNγ (25723.11, BD), CD58 (1C3, BD) and β2M (2M2, Biolegend). Cell viability was determined using propidium iodide exclusion or a fixable live/dead kit (Invitrogen). Intracellular staining assay (ICS) on ESO T cells after 5–6 hours of co-culture with A375 cells, where T cells were pre-treated with monensin (512092KZ, BD) and brefeldin A (512301KZ, BD), was performed using manufacturer’s instructions for BD Flow cytometry ICS. Flow cytometric data were acquired using either a FACSCanto II or LSRII Fortessa cytometer (BD), and data were analysed with FlowJo version 7.5 software (FlowJo LLC, Ashland, OR).
The amount of IFNγ release by T cells after co-culture with tumour cells was measured by Sandwich ELISA assay using anti-IFNγ (Thermo Scientific #M700A) coated 96-well plates, biotin-labeled anti-IFNγ (M701B), HRP-conjugated streptavidin (N100) and TMB substrate solution (N301).

Patel S.J., Sanjana N.E., Kishton R.J., Eidizadeh A., Vodnala S.K., Cam M., Gartner J.J., Jia L., Steinberg S.M., Yamamoto T.N., Merchant A.S., Mehta G.U., Chichura A., Shalem O., Tran E., Eil R., Sukumar M., Guijarro E.P., Day C.P., Robbins P., Feldman S., Merlino G., Zhang F, & Restifo N.P. (2017). Identification of essential genes for cancer immunotherapy. Nature, 548(7669), 537-542.

+ Open protocol

+ Expand

About PubCompare

Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.

We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.

However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.

Ready to get started?

Revolutionizing how scientists
search and build protocols!