Cd4 pe clone rm4 4

Manufactured by BioLegend

CD4-PE clone RM4–4 is a fluorescently labeled antibody that binds to the CD4 surface antigen. CD4 is a co-receptor expressed on the surface of T helper cells, playing a role in the activation and regulation of the immune response.

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4 protocols using cd4 pe clone rm4 4

Multiparametric Flow Cytometry Protocol

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CD3-FITC clone 145–2C11, CD4-PE-Cy7 clone RM4–5, CD4-PE clone RM4–4, CD11a-FITC clone M17/4, CD49d-PE clone R1–2, CXCR5-biotin clone L138D7, PE-Cy7 streptavidin, CD44-FITC and CD44-APC-Cy7 clone IM7, PD-1 APC clone 29F.1A12, GL7-PacBlue and GL7-PE clone GL7, CD19-PerCP-Cy5.5 clone 6B2, CD138-BV421 clone 281–2, IgD-APC-Cy7 clone 11–26c2a, CD95-PE clone SA367H8, CD73-BV605 clone TY/11.8, CD71-BV421 Clone RI7217, and CD80-PE clone 90, were purchased from Biolegend (San Diego, CA). BcL6-PE-CF594 clone K112–91 was purchased from BD Biosciences, San Jose, CA)

Waide M.L., Polidoro R., Powell W.L., Denny J.E., Kos J., Tieri D.A., Watson C.T, & Schmidt N.W. (2020). Gut Microbiota Composition Modulates the Magnitude and Quality of Germinal Centers during Plasmodium Infections. Cell reports, 33(11), 108503.

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Multiparametric Flow Cytometry Protocol

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Inducing Immune Tolerance in APP Mice

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To induce immune tolerance, APP mice were injected at days 0, 2 and 10 with 7.5mg/kg IP with GK1.5 anti-CD4 monoclonal antibody (Bioxcell). To confirm CD4 T-cell depletion, blood was taken on day 12 by retro orbital sampling into heparin coated tubes. CD4+ T lymphocytes were quantified using FACS analysis on a BD Fortessa using standard protocols with CD45-FITC (clone 104 BD Pharmigen), CD3e-AlexaFluor 647 (clone 17A2, eBioscience) and CD4-PE (RM4-4 clone, BioLegend) antibodies. GK1.5 treated animals had reduced CD4 as evidenced by a ratio of CD4+ lymphocytes/ total CD3+ lymphocytes of 0.04 +/- 0.008 (mean +/-SEM) in the treated mice compared to 0.47 +/- 0.003 from untreated mice.

Elmer B.M., Swanson K.A., Bangari D.S., Piepenhagen P.A., Roberts E., Taksir T., Guo L., Obinu M.C., Barneoud P., Ryan S., Zhang B., Pradier L., Yang Z.Y, & Nabel G.J. (2019). Gene delivery of a modified antibody to Aβ reduces progression of murine Alzheimer’s disease. PLoS ONE, 14(12), e0226245.

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Anti-CD4 Antibody Depletion in APP Mice

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The anti-CD4 monoclonal antibody GK1.5 was produced from a hybridoma acquired from American Type Culture Collection, ATCC® TIB-207 [38 (link), 39 (link)]. For immunodepletion induction, 11- to 12-week-old APP751SL mice were treated at day 0 and day 2 by intraperitoneal injection of 150 μg GK1.5 while a control group (i.e., non-immunotolerized mice) was administered with PBS. On day 4, blood was taken by retro orbital sampling on a heparin-lithium coated tube (microvette®, Sarstedt AG, Germany). CD4+ T lymphocytes were quantified using FACS analysis on a FACSCantoII flow cytometer using standard protocols with CD45-FITC (clone 104 BD Pharmigen), CD3e-alexa Fluor 647 (clone 17A2, eBioscience), and CD4-PE (RM4–4 clone, BioLegend) antibodies. All GK1.5-treated animals displayed reduced CD4 as evidenced by a ratio of CD4+ lymphocytes/total CD3+ lymphocytes of 0.01 ± 0.01 in the treated group compared with 0.48 ± 0.20 in the untreated mice.

Pradier L., Blanchard-Brégeon V., Bohme A., Debeir T., Menager J., Benoit P., Barneoud P., Taupin V., Bertrand P., Dugay P., Cameron B., Shi Y., Naimi S., Duchesne M., Gagnaire M., Weeden T., Travaline T., Reczek D., Khiroug L., Slaoui M., Brunel P., Fukuyama H., Ravetch J., Canton T, & Cohen C. (2018). SAR228810: an antibody for protofibrillar amyloid β peptide designed to reduce the risk of amyloid-related imaging abnormalities (ARIA). Alzheimer's Research & Therapy, 10, 117.

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