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Cd4 bv605

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The CD4 BV605 is a fluorochrome-conjugated antibody designed to detect and quantify CD4-positive cells in flow cytometry applications. It specifically binds to the CD4 surface antigen, which is expressed on a subset of T lymphocytes.

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Lab products found in correlation

Cd8 bv650, by BioLegend (4 mentions) Cd8 bv711, by BioLegend (2 mentions) Cd1d percp cy5, by BioLegend (2 mentions) Live dead fixable aqua dead cell stain kit, by Thermo Fisher Scientific (2 mentions) Cd3 af700, by BioLegend (2 mentions) Cd8 apc h7, by BD (2 mentions) Cd38 apc, by BD (2 mentions) Cd86 bv421, by BioLegend (2 mentions) Lsrfortessa, by BD (2 mentions) Cd3 fitc, by BioLegend (2 mentions) Lsr 2, by BD (2 mentions) Ebioscience foxp3 transcription factor staining buffer set, by Thermo Fisher Scientific (2 mentions) Facscanto 2, by BD (2 mentions) Facsdiva software, by BD (2 mentions) F4 80 bv421, by BioLegend (2 mentions) Cd3 bv570, by BioLegend (1 mentions) Cd4 bv605 rpa t4, by BD (1 mentions) Cd8 bv650 rpa t8, by BD (1 mentions) Horizon brilliant stain buffer, by BD (1 mentions) Live dead near ir, by Thermo Fisher Scientific (1 mentions)

Close spelling variants of this product

Anti-CD4 BV605 GK1.5 (2 citations) BV605 anti-mouse CD4 (4 citations) Anti-human CD4 BV605 (2 citations) Anti-CD4–BV605 (clone RM4-5, (2 citations) Anti-CD4 BV605 (RM4-5, (2 citations) BV605-conjugated anti-mouse CD4 (3 citations) Anti-CD4-BV605 (17 citations) Anti-human CD4-BV605 antibody (2 citations) CD4 PE-Cy7 (SK3); CD8 PerCP-Cy5.5 (RPA-T8); CCR7 FITC (G043H7); CD45RA BV421 (HI100); HLA-DR PE (L243); CD38 BV605 (HIT2) (2 citations) BV605 rat anti-mouse CD4 (1 citations)

22 protocols using cd4 bv605

1

Comprehensive Immunophenotyping of T Cells

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Cryopreserved peripheral blood mononuclear cells were thawed and stained in Horizon Brilliant Stain Buffer (BD) containing all antibodies and Live/Dead Near-IR (Life Technologies) at 1:300 dilution and stained at 4°C for 30 minutes in Horizon. Panel 1 included the following: CD3 BV570 (UCHT1), CCR7 Pacific Blue (G043H7), and CD27 AlexaFluor700 (M-T271) (all BioLegend); CD4 BV605 (RPA-T4) and CD8 BV650 (RPA-T8) (BD); programmed cell death protein 1 (PD-1) phycoerythrin (PE)–eFluor610 (eBioJ105), CD45RA fluorescein isothiocyanate (HI100), and T cell immunoreceptor with immunoglobulin and ITIM domains (TIGIT) peridinin-chlorophyll protein complex (PerCP)–eFluor710 (MBSA43) (eBioscience); and Tim-3 PE (344823) (R&D).
Panel 2 included the following: CD3 BV570, CD38 AlexaFluor700 (HB-7) (BioLegend); CD4 BV605, CD8 BV650, PD-1 PE-eFluor610, and Tim-3 PE. After this, cells were washed twice before fixation and permeabilization with Foxp3 Buffer Set (BD). Staining for intracellular epitopes was performed with: T-bet fluorescein isothiocyanate (4B10) (BioLegend) and Eomes eFluor660 (WD1928) (eBioscience). Samples were acquired on an LSR II flow cytometer (BD). Data were analyzed using FlowJo software (version 10.8.0r1; Tree Star).
Martin G.E., Pace M., Shearer F.M., Zilber E., Hurst J., Meyerowitz J., Thornhill J.P., Lwanga J., Brown H., Robinson N., Hopkins E., Olejniczak N., Nwokolo N., Fox J., Fidler S., Willberg C.B, & Frater J. (2019). Levels of Human Immunodeficiency Virus DNA Are Determined Before ART Initiation and Linked to CD8 T-Cell Activation and Memory Expansion. The Journal of Infectious Diseases, 221(7), 1135-1145.
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2

Naïve CD4+ T Cell Trafficking Modulation

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CD4 T cells were enriched using the naïve CD4 T cell isolation kit (Milteni Biotec, Cat. 130–104-453) from mononuclear cells isolated from the spleens and lymph nodes of congenically marked, wild-type (CD45.1) and CCR7-deficient (CD45.2) mice. A mixture of CD45.1-naïve CD4 T cells (3×106) and CD45.2-naïve CD4 T cells (1.5×106) were labeled with carboxyfluorescein succinimidyl ester (CFSE) and transferred into GF, B. thetaiotaomicron WT-monocolonized, or BtSULT Δsult-monocolonized B6 mice by retro-orbital injection. After 24h, recipient mice were sacrificed, and immune cells were isolated from the spleens and MLNs and stained with the following antibodies: CD45-Pacific blue (1:300, Biolegned), CD4-BV605 (1:300, Biolegend), CD25-PE (1:200, ebioscience), CD44-APC (1:300, ebiosience), CD62L-PE/Cy7 (1:300, Biolegend), CD45.1-PerCP/Cy5.5 (1:200, ebioscience) and CD45.2-APC/Fire750 (1:200, Biolegend). The ratio of donor CD4 T cells in either MLNs or spleens was first determined by dividing the number of CFSE+ CD4+ T cells (donor cells) by that of CD45+ cells (recipient cells) in each tissue. We then determine the relative migration of donor cells into the MLNs over the spleens of recipient mice by calculating the normalized ratio. Gating strategy and representative data are shown as Supplementary Figures 16b and 17c.
Helgason S., Petursson G., Gudmundsson S, & Sigurdsson J.A. (2000). Prevalence of postherpetic neuralgia after a first episode of herpes zoster: prospective study with long term follow up. BMJ : British Medical Journal, 321(7264), 794.
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3

Immune Phenotyping of PBMC Samples

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Cryopreserved peripheral blood mononuclear cell (PBMC) samples from baseline and post-treatment (cycles 1–12) were thawed and rested overnight at 37°C. Then cells were stained with a viability dye (eBioscience Fixable Viability Dye eFluor 450) and a master mix of antibodies for surface stains including CD4-BV605 (BioLegend Cat# 317438, RRID:AB_11218995), CD3-PE-Cyanine5.5 (Invitrogen Cat# 35-0036-42, RRID: AB_11220085), CD8a-PE-Cyanine7 (Invitrogen Cat# 25-0088-42, RRID: AB_1659702), PD1-APC (BioLegend Cat# 329908, RRID: AB_940475), CD152-PE-Cy5 (BD Biosciences Cat# 555854, RRID:AB_396177). Cells were next fixed and permeabilized with the eBioscience Foxp3/Transcription Factor Staining Buffer Set (ThermoFisher) and subsequently stained intracellularly with Alexa Fluor 700 antihuman Ki67 antibody (BioLegend Cat# 350530, RRID: AB_2564040). Stained cells were acquired on a BD Canto RUO and analyzed with FlowJo software (FlowJo, RRID:SCR_008520).
Liao J.B., Gwin W.R., Urban R.R., Hitchcock-Bernhardt K.M., Coveler A.L., Higgins D.M., Childs J.S., Shakalia H.N., Swensen R.E., Stanton S.E., Tinker A.V., Wahl T.A., Ancheta R.G., McGonigle K.F., Dai J.Y., Disis M.L, & Goff B.A. (2021). Pembrolizumab with low-dose carboplatin for recurrent platinum-resistant ovarian, fallopian tube, and primary peritoneal cancer: survival and immune correlates. Journal for Immunotherapy of Cancer, 9(9), e003122.
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4

Multi-color Flow Cytometry Immunophenotyping

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For direct multi-colour flow cytometry (LRS Fortessa X-20; BD Bioscience), cells were incubated for 30 min at 4 °C with the antibodies mentioned below. Live/dead cell discrimination was performed by staining with the Zombie Aqua Fixable Viability Kit (BioLegend, catalogue number: 423101). To block Fc receptors, a purified anti-mouse CD16/CD32 (eBioscience, catalogue number: 14-0161-86) was used.
CD19 APCCy7 (clone 6D5, Biolegend, catalogue number: 115530, 1:300), Ly6G PerCPCy5.5 (clone 1A8, Biolegend, catalogue number: 127616, 1:300), CD45 PEDazzle (clone 30-F11, Biolegend, catalogue number: 103146, 1:400), F4/80PECy7 (clone BM8, eBioscience, catalogue number:254801-82, 1:200), CD3 APC (clone 145-2c11, Biolegend, catalogue number: 100312, 1:100), CD11b Alexa700 (clone M170, Biolegend, catalogue number: 101222, 1:400), TCγδV421 (clone GL3, Biolegend, catalogue number: 118120, 1:100), CD4 BV605 (clone RM4-5, Biolegend, catalogue number: 100548, 1:300), CD8 BV711 (clone 53-6.7, Biolegend, catalogue number: 100748, 1:300) and NK1.1 PE (clone PK136, Biolegend, catalogue number: 557391, 1:300). IL-10GFP production was measured in the FITC channel.
Mukherjee D., Chora Â.F., Lone J.C., Ramiro R.S., Blankenhaus B., Serre K., Ramirez M., Gordo I., Veldhoen M., Varga-Weisz P, & Mota M.M. (2022). Host lung microbiota promotes malaria-associated acute respiratory distress syndrome. Nature Communications, 13, 3747.
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5

Multiparameter Analysis of Immune Cell Subsets

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Single-cell suspensions from spleens, bone marrow, or peripheral lymph nodes were blocked for 30 min using FBS. Cell-surface staining was achieved by incubating cells at 4°C for 30 min with fluorescence-conjugated anti–mouse antibodies (clone; source): XBP1s–Alexa Fluor 647 (Q3-695; BD), XBP1s-phycoerythrin (PE; Q3-695; BD), B220–Alexa Fluor 488 (RA3-6B2; BioLegend), B220-BV605 (RA3-6B2; BioLegend), CD43-PE (eBioR2/60; eBioscience), CD19–Alexa Fluor 647 (6D5; BioLegend), IgM-PE-Cy7 (RMM-1; BioLegend), IgD-FITC (11-26c.2a; BioLegend), GL7-PE (GL7; BioLegend), AA4.1-PE-Cy7 (AA4.1; BioLegend), CD1d-PerCP-Cy5.5 (1B1; BioLegend), CD23-FITC (B3B4; BioLegend), CD3-APC-Cy7 (145-2C11; BioLegend), CD4-BV605 (RM4-5; BioLegend), CD8α-PE-Cy7 (53–6.7; BioLegend), and CD138-PE (281–2; BioLegend). Viability staining was accomplished using DAPI exclusion during acquisition. Acquisition of B, T, and dendritic cell populations was performed on an LSRII cytometer (BD) harboring a custom configuration for the Wistar Institute. Cytometry data were analyzed using FlowJo software (7.6.1; Tree Star Inc.).
Tang C.H., Chang S., Paton A.W., Paton J.C., Gabrilovich D.I., Ploegh H.L., Del Valle J.R, & Hu C.C. (2018). Phosphorylation of IRE1 at S729 regulates RIDD in B cells and antibody production after immunization. The Journal of Cell Biology, 217(5), 1739-1755.
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6

Phenotyping and Proliferation of Anti-SHIV T Cells

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Surface markers and intracellular Ki67were stained for phenotyping and measuring in vivo adoptively transferred and proliferating anti-SHIV T Cells. After PBMCs were isolated from the blood and tissue samples, cells were washed twice with PBS and stained with LIVE/DEAD® Fixable Aqua Dead Cell Stain Kit (Thermo Fisher SCIENTIFIC) for 20 min at room temperature followed by staining with the following antibodies: CD3-Cy7-APC (Clone: SP34.2, BD Biosciences), CD4-BV605 (Clone: OKT4, BioLegend), CD8-BV711 (Clone: RPA-T8, BioLegend), CD28-Cy5PE (Clone: 28.2, BD Biosciences), CD45RA-Cy7-PE (Clone: L48, BD Pharmingen), CD69-ECD (Clone: TP1.55.3, Beckman Coulter), CCR7-Alexa Flour 680 (Clone: 150503, BD Pharmingen) and HLA-DR-BV421 (Clone: LN3, Biolegend). After surface staining, cells were permeabilized with Foxp3 / Transcription Factor Staining Buffer Set (eBiosciences) for 20 min at room temperature, then intracellular stained with anti-Ki67-PE (Clone: B56, BD Pharmingen).
Iwamoto N., Patel B., Song K., Mason R., Bolivar-Wagers S., Bergamaschi C., Pavlakis G.N., Berger E, & Roederer M. (2021). Evaluation of chimeric antigen receptor T cell therapy in non-human primates infected with SHIV or SIV. PLoS ONE, 16(3), e0248973.
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7

Immunophenotyping of Whole Blood and PBMCs

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For immunofluorescence staining of whole blood and PBMC, antibodies included CD45-ECD (Beckman Coulter, Indianapolis IN), CD3-e780, CD4-e450, CD8-AF700, CD15-FITC (eBioscience, San Diego, CA), CD45-FITC, CD4-PerCP-Cy5-5, CD14-PECy7, HLA-DR-APC, HLA-DR-BV421, CD8-APC-H7, CCR7-BV605, CD25-BV605, CD45RA-PECy7, CD45RO-APC-H7, CCR4-PECy7, CD38-APC, CD127-BV650 (BD Bioscience, San Jose, CA), CD14-AF700, and CD3-AF700 (Biolegend, San Diego, CA). For phosphoSTAT staining, antibodies included CD3-BV785 (Biolegend), CD4-BV605, CD8-BV510, CD14-PECy7, CD19-BV421, CD16-BV650, pSTAT3-647, pSTAT5-PE (BD Biosciences), and pSTAT1-488 (Cell Signaling Technologies, Danvers, MA). Cell types were identified according to the criteria of the Human Immunology Project [19 (link)], and a full list of antibodies used to identify individual cell types including those not reported here can be found in Additional file 2: Table S1. Hematological toxicity was graded according to NCI CTCAE v4.0 guidelines and is reported in Additional files 3 and 4: Tables S2 and S3.
Crocenzi T., Cottam B., Newell P., Wolf R.F., Hansen P.D., Hammill C., Solhjem M.C., To Y.Y., Greathouse A., Tormoen G., Jutric Z., Young K., Bahjat K.S., Gough M.J, & Crittenden M.R. (2016). A hypofractionated radiation regimen avoids the lymphopenia associated with neoadjuvant chemoradiation therapy of borderline resectable and locally advanced pancreatic adenocarcinoma. Journal for Immunotherapy of Cancer, 4, 45.
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8

Characterization of Immune Cell Activation

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In order to examine activation status of the cells, BMDCs were treated with Duolac ATP or LPS for 24 h at 37°C. The cells were stained with anti-mouse CD86-FITC, PD-L1-PE, MHC II-PE-cy7, CD11c-APC (all from BD Biosciences) for 20 min at 4°C in the dark. To test the increase of Treg, CTV labeled CD4+ T cells cultured with Duolac-treated BMDCs for 3 days were stained with anti-mouse CD4-PE (BD Biosciences). After surface staining, CD4+ T cells were fixed and stained with anti-mouse Foxp3-APC mAb (BioLegend, Dedham, MA, United States) using FOXP3 Fix/Perm Buffer Set (BioLegend). In vivo examination, single cells from mLNs and PP were isolated from AD mice. Population changes of DCs and Tregs were examined as aforementioned. To analyze for subpopulation of Th cells, total mLN and PP cells were stimulated with PMA and ionomycin (Sigma-Aldrich) in the presence of brefeldin A for 4 h. After the stimulation, the cells were stained with appropriate combination of anti-mouse CD11c-APC, CD4-bv605, Foxp3-APC, IFN-gamma-PE, IL-4-bv605, and IL-17-APC-cy7 mAb (all from Biolegend). The cells were washed and the expression was examined using a FACSCanto II (BD Biosciences). All flow cytometric data acquired were analyzed with FlowJo software (Tree Star, Ashland, OR, United States).
Kim H.W., Hong R., Choi E.Y., Yu K., Kim N., Hyeon J.Y., Cho K.K., Choi I.S, & Yun C.H. (2018). A Probiotic Mixture Regulates T Cell Balance and Reduces Atopic Dermatitis Symptoms in Mice. Frontiers in Microbiology, 9, 2414.
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9

Tumor Immune Cell Profiling

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Tumors were harvested, weighed, and processed to obtain single-cell suspensions. In brief, tumor tissues were dissociated and digested with 250 µg/mL of Liberase (Roche) and incubated for 30 min at 37 °C, while shaking at 105 rpm for proper digestion. Fetal bovine serum was then added to stop the reaction, and samples were filtered and washed with PBS + 2% FBS. The cell count per sample was performed, then samples were stained using fluorochrome-conjugated antibodies from BioLegend including: CD45 Pacific blue, cat# 103126; CD4 BV605, cat# 100451; CD8 PE, cat# 100707; CD49b APC, cat# 108910; Foxp3 Alexa488, cat# 126406; TIGIT PE-Cy7, cat# 142108; Gr1 BV510, cat# 108437; CD11b APC-fire750, cat# 101262; F4/80 Alexa700, cat# 123130; CD206 PercpCy5.5, cat# 141716. In alternate panels, a set of other antibodies was also used including: CD4 FITC, cat# 100406; CD8 PErcpCy5.5, cat# 100734; Gr1 APC, cat# 108412; CD11c BV510, cat# 117337; and PVR (CD155) PE, cat# 132206. Samples were run on the Attune flow cytometer, and data were analyzed using Flow-Jo 10 software.
Barsoumian H.B., Sezen D., Menon H., Younes A.I., Hu Y., He K., Puebla-Osorio N., Wasley M., Hsu E., Patel R.R., Yang L., Cortez M.A, & Welsh J.W. (2022). High Plus Low Dose Radiation Strategy in Combination with TIGIT and PD1 Blockade to Promote Systemic Antitumor Responses. Cancers, 14(1), 221.
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10

Modulation of Gut Immunity in Peanut Allergy

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Peanut-sensitized mice were intragastrically administered twice daily with PBS, sodium butyrate or ButM at an equivalent dose of 0.224 mg g−1 (butyrate per mouse) for 2 weeks. After the treatment, spleen, ileum and colon-draining LNs from mice were collected, and digested in DMEM supplemented with 5% FBS, 2.0 mg ml−1 collagenase D (Sigma Aldrich) and 1.2 ml CaCl2. Single-cell suspensions were prepared by mechanically disrupting the tissues through a cell strainer (70 μm, Thermo Fisher). Splenocytes (4 × 106) or cells from LNs (1 × 106) were plated in a 96-well plate. Cells were stained with LIVE/DEAD Fixable Aqua Dead Cell Stain kit (Thermo Fisher), followed by surface staining with antibodies in PBS with 2% FBS, and intracellular staining according to the manufacturer’s protocols for eBioscience Foxp3/Transcription Factor Staining Buffer set (Invitrogen). The following anti-mouse antibodies were used: CD3 APC/Cy7 (clone 145-2C11, BD Biosciences), CD4 BV605 (clone RM4-5, Biolegend), CD25 PE/Cy7 (clone PC61, Biolegend), Foxp3 AF488 (clone MF23, BD Biosciences), CD11b BV711 (clone M1/70, BD Biosciences), CD11c PE/Cy7 (clone HL3, BD Biosciences), F4/80 APC (clone RM8, Biolegend), I-A/I-E (MHCII) APC/Cy7 (clone M5/114.15.2, Biolegend) and CD86 BV421 (clone GL-1, Biolegend). Stained cells were analysed using an LSR Fortessa flow cytometer (BD Biosciences).
Wang R., Cao S., Bashir M.E., Hesser L.A., Su Y., Hong S.M., Thompson A., Culleen E., Sabados M., Dylla N.P., Campbell E., Bao R., Nonnecke E.B., Bevins C.L., Wilson D.S., Hubbell J.A, & Nagler C.R. (2022). Treatment of peanut allergy and colitis in mice via the intestinal release of butyrate from polymeric micelles. Nature Biomedical Engineering, 7(1), 38-55.
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