Cd4 rm4 5

Manufactured by BD

Sourced in United States

The CD4 (RM4-5) is a lab equipment product designed to measure the presence and quantity of CD4 cells in a biological sample. It serves as a core function in various medical and research applications.

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Lab products found in correlation

Cd8 53 6, by BD (7 mentions) Cd11b m1 70, by BD (7 mentions) Cd44 im7, by BD (6 mentions) Cd3 145 2c11, by BD (4 mentions) Cd62l mel 14, by BD (4 mentions) Ly6g 1a8, by BioLegend (4 mentions) Foxp3 fjk 16s, by Thermo Fisher Scientific (4 mentions) B220 ra3 6b2, by BD (4 mentions) Tcrβ h57 597, by Thermo Fisher Scientific (3 mentions) Phorbol myristate acetate (pma), by Merck Group (3 mentions) Ionomycin, by Merck Group (3 mentions) Ly6c al 21, by BD (3 mentions) Tc11 18h10, by BD (3 mentions) F4 80 bm8, by Thermo Fisher Scientific (3 mentions) Ly6c hk1, by BioLegend (3 mentions) Cd11c hl3, by BD (3 mentions) Siglecf e50 2440, by BD (3 mentions) Cd45 30 f11, by BD (3 mentions) Fc block, by BD (3 mentions) Cd25 pc61, by Thermo Fisher Scientific (3 mentions)

Close spelling variants of this product

RM4-5 (42 citations) Anti-CD4 (RM4-5, (25 citations) Anti-mouse CD4 (clone RM4-5, (6 citations) CD4-APC (RM4-5) (3 citations) Anti-CD4-FITC (RM4-5, (3 citations) Anti‐CD4 PerCP (clone RM4‐5) (2 citations) CD4 BV786 (RM4-5 (2 citations) Anti-CD4-Pacific Blue and anti-CD4-FITC (clone RM4-5, (2 citations) CD4 PE (RM4-5; (2 citations) CD4-peridinin chlorophyll protein (PerCP).Cy5.5 (RM4-5; (2 citations)

35 protocols using cd4 rm4 5

Multiparametric Analysis of Immune Cells

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Flow cytometry and cell sorting were completed on a FACSAria Fusion instrument (BD) and analyzed using FlowJo analysis software (Tree Star). Surface staining was performed at 4°C in the presence of Fc block (2.4G2) in magnetic-activated cell sorting (MACS) buffer (PBS, 0.5% BSA, 2 mM EDTA). Intracellular IRF8 staining was performed using the Foxp3 staining kit (eBioscience 00-5523-00).
The following antibodies were used: CD19 (1D3), CD135 (A2F10.1), MHCII (M5/114.15.2), CD117 (2B8), B220 (RA3-6B2), CD3 (145-2C11), and CD4 (RM4-5) from BD Biosciences; CD3 (145-2C11), CD4 (GK1.5), and MHCII (M5/114.15.2) from Tonbo Biosciences; TER-119 (TER-119), Ly-6G (1A8), B220 (RA3-6B2), CD24 (M1/69), CD115 (AFS98), XCR1 (ZET), CD19 (6D5), CD8α (53-6.7), CD4 (RM4-5), CD11c (N418), and CD3 (17A2) from Biolegend; CD105 (MJ7/18), Siglec-H (eBio440c), CD3 (17A2), CD8α (53-6.7), CD11c (N418), and IRF8 (V3GYWCH) from eBiosciences; and SA-Qdot 605, CD11c (N418), CD317 (eBio927), CD172a (P84), and TCRβ (H57-597) from Invitrogen.

Liu T.T., Ou F., Belk J.A., Bagadia P., Anderson DA I.I.I., Durai V., Yao W., Satpathy A.T., Murphy T.L, & Murphy K.M. (2023). Cis interactions in the Irf8 locus regulate stage-dependent enhancer activation. Genes & Development, 37(7-8), 291-302.

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Comprehensive Lymphocyte Immunophenotyping

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Lymph nodes and spleen were disintegrated into single cells as previously described45 (link) and stained for surface markers with the following antibodies: TCR-β (H57-597), B220 (R43-6B2), CD4 (RM4-5), and CD8 (53-6.7), and all were purchased from BD Biosciences (Franklin Lakes, NJ, USA) or eBioscience (San Diego, CA, USA).
Spleen cells were stimulated as previously described46 , and then stained for intracellular cytokines using a Mouse FOXP3 Buffer Set (BD Biosciences). The following fluorochrome-conjugated antibodies were used: CD4 (GK1.5), CD25 (7D4), CD3 (145-2C11), FOXP3 (FJK-16s), RORγt (AFKJ-9), IFNγ (XMG1.2), and IL-17 (eBio17B7), and all were purchased from BD Biosciences or eBioscience.
Data was collected with a FACSCanto instrument (BD Biosciences).

Zhao M., Chen H., Ding Q., Xu X., Yu B, & Huang Z. (2016). Nuclear Factor Erythroid 2-related Factor 2 Deficiency Exacerbates Lupus Nephritis in B6/lpr mice by Regulating Th17 Cell Function. Scientific Reports, 6, 38619.

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Murine Lung Immune Cell Profiling

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P1 and P21 male and female murine lungs were isolated as described above. Cells were blocked for 30 min with CD16/CD32 (Tonbo Biosciences). For intracellular analyses, cells were stimulated with PMA (50 ng/ml, Sigma-Aldrich), ionomycin (750 ng/mL, Sigma-Aldrich), and GolgiStop (BD Biosciences) for 5 hr, and then surface stained with fluorochrome-conjugated antibodies for 30 min: CD3 (clone: 145–2 C11, BD Biosciences), CD4 (RM4-5, BD Biosciences), and CD8a (clone: 53–6.7, Biolegend). Cells were then permeabilized with FOXP3 Fixation/Permeabilization Kit (BD Biosciences) as indicated by the manufacturer, and stained for TBET (clone: 4B10, Biolegend), GATA3 (clone: L50-823, BD Biosciences), FOXP3 (clone: FJK-16s, eBioscience), RORγt (clone: Q31-378, BD Biosciences), IFNγ (clone: XMG1.2, Biolegend), IL-4 (clone: 11B11, BD Biosciences), IL-10 (clone: JES5-16E3, Biolegend), and IL-17 (clone: TC11-18H10, Miltenyi Biotec) for 30 min. Cells were read using an LSRII flow cytometer using FACSDiva software. Flow data was analyzed using FlowJo (Tree Star Inc). Flow cytometry analysis for this project was done on instruments in the Stanford Shared FACS Facility.

Domingo-Gonzalez R., Zanini F., Che X., Liu M., Jones R.C., Swift M.A., Quake S.R., Cornfield D.N, & Alvira C.M. (2020). Diverse homeostatic and immunomodulatory roles of immune cells in the developing mouse lung at single cell resolution. eLife, 9, e56890.

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Comprehensive Antibody Panel for T-Cell Analysis

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The following antibodies were used from eBioscience (San Diego, CA) unless otherwise specified: Helios (22F6; Biolegend, San Diego, CA), CD90.1 (H1S51), TCRβ (H57-597; BD Biosciences, San Jose, CA), CD69 (H1.2F3), Vβ5 (MR9-4; BD Biosciences), Foxp3 (FJK16S), CD45RB (C363.16A), CD45.1 (A20), Vα2 (B20.1), CD3ε (eBio500A2), CD152 (UC10-4B9), folate receptor 4 (eBio12A5), CD73 (eBioTY/11.8), Vα2 (B20.1; BD Biosciences), CD25 (PC61.5), and CD4 (RM4-5; BD Biosciences).

Geem D., Ngo V., Harusato A., Chassaing B., Gewirtz A.T., Newberry R.D, & Denning T.L. (2016). Contribution of Mesenteric Lymph Nodes and GALT to the Intestinal Foxp3+ Regulatory T-Cell Compartment. Cellular and Molecular Gastroenterology and Hepatology, 2(3), 274-280.e3.

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Comprehensive Multiparameter Flow Cytometry

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The Cyan ADP flow cytometer (Beckman Coulter, Indianapolis IN) was used for all flow samples. Antibodies for these surface markers were used for flow cytometry: CD3 (145-2C11, BD Bioscience Franklin Lakes, NJ, USA), CD4 (RM4-5, BD Bioscience), anti-CEA CAR (Wi2, Immunomedics Morris Plains, NJ, USA), CD11b (M1/17, BD Bioscience), Ly6C (AL-21, BD Bioscience), Ly6G (1AB, BD Bioscience), PD-L1 (MIH5, BD Bioscience), CD62L (MEL-14, BD Bioscience), CCR7 (4B12, BD Bioscience), CD44 (IM7, BD Bioscience). Intracellular FoxP3 staining was performed with Mouse FoxP3 Permeabilization Kit (BD Bioscience). Single stain and isotype controls were used for each experiment. Analysis of acquired flow samples was performed with FlowJo software (Tree Star Inc., Ashland OR).

Katz S., Point G.R., Cunetta M., Thorn M., Guha P., Espat N.J., Boutros C., Hanna N, & Junghans R.P. (2016). Regional CAR-T cell infusions for peritoneal carcinomatosis are superior to systemic delivery. Cancer gene therapy, 23(5), 142-148.

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Skin Allograft Immune Profiling by Flow Cytometry

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Flow cytometric analysis of skin allograft, draining lymph node (DLN) and spleen was performed, and each leukocyte population was quantified. CD4 (RM4-5, #100559, 1:400), CD8 (53-6.7, #100734, 1:400), CD69, CD25 (PC61.5, #12-0251-81B, 1:400), Foxp3 (FJK-16s, #11-5773-82, 1:300) and Annexin-PE (51-65875X) were purchased from BD Biosciences, San Jose, CA. For intracellular cytokine staining, cells were stimulated ex vivo with phorbol 12-myristate 13-acetase (PMA, 50 ng/ml) and ionomycin (500 ng/ml) in combination with GolgiStop for 4 hours, then permeabilized and stained with fluorochrome conjugated antibodies against IFNγ. Cells were run on FACSCanto II (BD Biosciences, Franklin Lakes, NJ). Data were analyzed using FlowJo software.

Uehara M., Li X., Sheikhi A., Zandi N., Walker B., Saleh B., Banouni N., Jiang L., Ordikhani F., Dai L., Yonar M., Vohra I., Kasinath V., Orgill D.P., Khademhosseini A., Annabi N, & Abdi R. (2019). Anti-IL-6 eluting immunomodulatory biomaterials prolong skin allograft survival. Scientific Reports, 9, 6535.

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Multiparametric Flow Cytometry Analysis

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All flow cytometric analyses and sorting were performed on a BD FACS Aria II cell sorter. Antibodies used were CD4 (RM4-5, BD Biosciences), CD8a (53-6.7, eBioscience), B220 (RA3-6B2, Biolegend), CD11b (M1/70, BD Biosciences), Ly6G/C (RB6-8C5, Biolegend), Ter119 (TER-119, Biolegend), Sca1 (D7, Biolegend), MPL (AMM2, Immuno-Biological Laboratories), CD41 (MWReg30, BD Biosciences), CD150 (TC15-12F12.2, Biolegend), cKit (2B8, Biolegend), CD34 (RAM34, eBioscience), IL7R (A7R34, eBioscience), Flt3 (A2F10, eBioscience), CD16/32 (93, eBioscience) and CD105 (MJ7/18, Biolegend). Streptavidin (eBioscience) was used to resolve biotinylated antibodies and propidium iodide was added prior to analysis to identify live cells.

O’Neill A., Chin D., Tan D., Majeed A.B., Nakamura-Ishizu A, & Suda T. (2020). Thrombopoietin maintains cell numbers of hematopoietic stem and progenitor cells with megakaryopoietic potential. Haematologica, 106(7), 1883-1891.

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Immunohistochemical Analysis of Lymphoid Tissues

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Immunohistochemistry was performed as described (15 (link)). Briefly, freshly isolated LNs or spleens were fixed in newly prepared 4% paraformaldehyde (Electron Microscopy Science) overnight at 4 °C on an agitation stage. Spleens or LNs were embedded in 4% low melting agarose (Invitrogen) in PBS and sectioned with a vibratome (Leica VT-1000 S) at a 30 μm thickness. Thick sections were blocked in PBS containing 10% fetal calf serum, 1 mg/ml anti-Fcγ receptor (BD Biosciences), and 0.1% Triton X-100 (Sigma) for 30 minutes at room temperature. Sections were stained overnight at 4 °C on an agitation stage with antibodies directed at B220 (RA3–6B2, BD Biosciences), CD4 (RM4–5, BD Biosciences), CD169 (3D6.112), CD21/35 (4E3), CD45.1 (A20), CD45.2(104) all from BioLegend, Ki67 (SolA15, ThermoFisher), and LYVE-1 (223322, R & D Systems). Stained thick sections were microscopically analyzed using a Leica SP8 confocal microscope equipped with an HC PL APO CS2 40× (NA, 1.30) oil objective (Leica Microsystem, Inc.) and images were processed with Leica LAS AF software (Leica Microsystem, Inc.) and Imaris software v.9.0.1 64× (Bitplane AG). The intensities of fluorescent signals in regions of interests (ROI) were measured by LSA AF Lite software (Leica Microsystem).

Hwang I.Y., Park C., Harrison K, & Kehrl J.H. (2019). Biased S1PR1 Signaling in B cells Subverts Responses to Homeostatic Chemokines Severely Disorganizing Lymphoid Organ Architecture. Journal of immunology (Baltimore, Md. : 1950), 203(9), 2401-2414.

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Isolation and Flow Cytometry of Murine Intestinal Lamina Propria Cells

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The following antibodies were purchased from eBioscience: IFNγ (XMG1.2), CD90.1 (H1S51), CD69 (H1.2F3), CD45RB (C363.16A), CD45.1 (A20), Vα2 (B20.1), IL-17A (eBio17B7), CD8α (eBioT4/11.8), CD25 (PC61.5), CD3ε (eBio500A2), and RORγ(t)-PE (B2D). Antibodies purchased from BD Biosciences were: TCRβ (H57-597), Vβ5 (MR9-4), CCR6 (140706), IL-17A (TC11-18H10), Vα2 (B20.1), and CD4 (RM4-5). Dead cells were identified using the fixable Aqua dead cell staining kit (Invitrogen). The following biotin-conjugated antibodies (eBioscience) were used for negative selection in conjunction with anti-biotin and anti-APC microbeads (Miltenyi Biotec): CD8α (53-6.7), Ly-6G (RB6-8C5), F4/80 (BM8), TER-119 (TER-119), CD11b (M1/70), NK1.1 (PK136), CD11c (N418), CD19 (eBio1D3). Isolation of LP cells and flow cytometry was performed as previously described (18 (link)).

Geem D., Medina-Contreras O., McBride M., Newberry R.D., Koni P.A, & Denning T.L. (2014). Specific Microbiota-Induced Intestinal Th17 Differentiation Requires MHC II but not GALT and Mesenteric Lymph Nodes. Journal of immunology (Baltimore, Md. : 1950), 193(1), 431-438.

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Multiparameter flow cytometry analysis

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Single cell suspensions were stained with a viability dye, followed by incubation with purified rat anti-mouse CD16/CD32 (mouse BD Fc Block, clone 2.4G2) as previously described (22 (link)). Cells were then stained for 25 min at 4°C in the dark using antibodies targeting the following markers: CD69 (H1.2F3, Biolegend), CD4 (RM4-5, BD Biosciences), CD11a (2D7, BD Biosciences), and CD44 (IM7, Tonbo), (MEL-14, Biolegend). Cells were then washed and prepared for flow cytometry analysis. Data was acquired using a BD LSR-II instrument, configured with 488 (blue), 633 (red), 407 (violet), and 532 (green)-nm lasers. Data were analyzed using Flowjo software (FlowJo, LLC), version 10.

Richards K.A., DiPiazza A.T., Rattan A., Knowlden Z.A., Yang H, & Sant A.J. (2018). Diverse Epitope Specificity, Immunodominance Hierarchy, and Functional Avidity of Effector CD4 T Cells Established During Priming Is Maintained in Lung After Influenza A Virus Infection. Frontiers in Immunology, 9, 655.

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