Pecy5 anti cd4

Manufactured by BioLegend

Sourced in United States

PECy5 anti-CD4 is a fluorochrome-conjugated monoclonal antibody that binds to the CD4 surface antigen. It is commonly used in flow cytometry applications to identify and study CD4+ T cells.

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Lab products found in correlation

Facsaria sorp, by BD (2 mentions) Anti cd8 pe cy7, by BioLegend (2 mentions) Bovine serum albumin (bsa), by Carl Roth (2 mentions) Bv510 anti b220, by BioLegend (2 mentions) Percpcy5.5 anti cd11b, by BioLegend (2 mentions) Lsrii system, by BD (1 mentions) Lymphocyte separation medium, by PromoCell (1 mentions) Anti cd3 pe, by BD (1 mentions) Anti mouse cd16 cd32, by BD (1 mentions) Anti cd8 pe cy7, by Thermo Fisher Scientific (1 mentions) Facscanto flow cytometer, by BD (1 mentions) Anti foxp3 alexa488, by Thermo Fisher Scientific (1 mentions) Clone 53 6, by BioLegend (1 mentions) Clone h129, by BioLegend (1 mentions) Clone rb6 8c5, by BioLegend (1 mentions) Clone ra3 6b2, by BioLegend (1 mentions) Clone m1 70, by BioLegend (1 mentions) Penicillin streptomycin pen strep, by Thermo Fisher Scientific (1 mentions) Anti cd45 apc cy7, by BioLegend (1 mentions) Anti cd11c bv421, by BioLegend (1 mentions)

Close spelling variants of this product

Anti-CD4 (192 citations) PeCy5.5 (2 citations)

5 protocols using pecy5 anti cd4

Characterization of Colonic Foxp3+ T Cells

Cited 1 time

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The colonic tissues from Foxp3-GFP mice were prepared for processing single-cell suspensions of lymphocytes from colonic lamina propria, as described previously.^{18 (link)} Cells were labeled with LIVE/DEAD fixable viability dyes (#2443413, Invitrogen, Waltham, MA) and PE-Cy5-anti-CD4 (#100410, BioLegend, San Diego, CA) at room temperature. Then, cells were analyzed using multi-color flow cytometry, a BD LSRII system (BD Biosciences) to determine the percentage of GFP (Foxp3 expression) and PE-Cy5.5 (CD4 expression) double positive cells within live cell population. Each sample contained lymphocytes from 2 to 3 mice with the same treatment.

Kaur H., Ali S.A., Short S.P., Williams C.S., Goettel J.A., Washington M.K., Peek RM J.r., Acra S.A, & Yan F. (2023). Identification of a functional peptide of a probiotic bacterium-derived protein for the sustained effect on preventing colitis. Gut Microbes, 15(2), 2264456.

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Immune Cell Profiling in Tamoxifen-Treated Mice

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EDTA-blood was collected from the orbital sinus 2 weeks after last tamoxifen injection. Per mouse, 50µL blood were diluted in 50µL PBS and overlaid on 100µL lymphocyte separation medium (1077, PromoCell). After centrifugation, peripheral blood mononuclear cells were isolated, washed and stained for 15 min at 4 °C with the following antibodies: PECy5 anti-CD4 (1:1000, clone H129.19, Biolegend), PECy7 anti-CD8 (1:500, clone 53–6.7, Biolegend), BV510 anti-B220 (1:500, clone RA3-6B2, Biolegend) and PerCpCy5.5 anti-CD11b (1:100, clone M1/70, Biolegend). After staining, cells were washed, suspended in 200µL PBS containing 2% bovine serum albumin (#8076.3, Roth), and filtered through 40 µm cell strainers. Samples were measured on a FACSAria Sorp (BD). Number of lymphocytes were determined using forward and side scatter. Frequency of T-helper cells (CD4⁺,CD8⁻), cytotoxic T-cells (CD8⁺, CD4⁻), B cells (B220⁺, CD11b⁻, CD8⁻, CD4⁻), and myeloid cells (CD11b⁺, B220⁻, CD8⁻, CD4⁻) were determined as percentage of total lymphocytes.

Wilke J.B., Hindermann M., Moussavi A., Butt U.J., Dadarwal R., Berghoff S.A., Sarcheshmeh A.K., Ronnenberg A., Zihsler S., Arinrad S., Hardeland R., Seidel J., Lühder F., Nave K.A., Boretius S, & Ehrenreich H. (2021). Inducing sterile pyramidal neuronal death in mice to model distinct aspects of gray matter encephalitis. Acta Neuropathologica Communications, 9, 121.

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Comprehensive Immune Cell Analysis in Murine Tumor Models

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The immune phenotype of splenocytes and tumor infiltrating lymphocytes (TIL) were analyzed in all the three models by flow cytometry. TILs were isolated as previously described [12 (link)]. Splenocytes were processed for analysis according to our previously published methods [13 (link)]. For FOXP3 and CD4/CD8 evaluation, 1μg anti-mouse CD16/CD32 (BD Pharmingen) was used to block nonspecific binding for 30 minutes at room temperature then 0.4μg each of PE anti-CD3 (BD Pharmingen, clone #145-2C11), PE/Cy5 anti-CD4 (BioLegend, clone #GK1.5), and PE-Cy7 anti-CD8 (eBioscience, clone #53-6.7) were added. After overnight permeabilization of the cells, 1μg anti-FOXP3 Alexa488 (eBioscience, clone #FJK.16s) was added for 30 minutes. For myeloid derived suppressor cell (MDSC) evaluation, 0.4μg of GR-1 (BD Pharmingen, clone #RB6-8C5) and anti-mouse CD11b were added and incubated for 30 minutes at room temperature (eBioscience clone #M1/70). After the appropriate antibodies were applied to each single cell suspension of tumor or spleen for 30 min at room temperature, the stained cells were acquired with FACSCanto flow cytometer (BD Bioscience) and analyzed with FlowJo software (Tree Star Inc.). Results are reported as mean ± SEM of the total percentage of a cell population or ratio of cell quantities, as indicated.

Gad E., Rastetter L., Slota M., Koehnlein M., Treuting P.M., Dang Y., Stanton S, & Disis M.L. (2014). Natural history of tumor growth and immune modulation in common spontaneous murine mammary tumor models. Breast cancer research and treatment, 148(3), 501-510.

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Multiplex Flow Cytometry of Immune Cells

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Per mouse, 50 µL EDTA blood was diluted in 50 µL PBS and overlaid on 100 µL lymphocyte separation medium (1077, PromoCell). After centrifugation, cells were isolated from the interphase, washed and stained for 15 min at 4 °C with the following antibodies: PECy5 anti-CD4 (1:1000, clone H129.19, BioLegend), PECy7 anti-CD8 (1:500, clone 53-6.7, BioLegend), BV510 anti-B220 (1:333, clone RA3-6B2, BioLegend), PerCpCy5.5 anti-CD11b (1:1000, clone M1/70, BioLegend), PE anti-Gr1 (1:1000, clone RB6-8C5, BioLegend), and FITC anti-F4/80 (1:1000, BM8, BioLegend). After staining, cells were washed, suspended in 250 µL PBS containing 2% bovine serum albumin (#8076.3, Roth), and filtered through 40 µm cell strainers. Samples were measured on a FACSAria Sorp (BD). Total cell numbers were determined using forward and side scatter. Frequency of helper T cells (CD4+, CD8−), cytotoxic T cells (CD8+, CD4−), B cells (B220+), macrophages (CD11b+, F4/80+) and neutrophils/monocytes (CD11b+, Gr1+) were determined as percentage of total lymphocytes.

Arinrad S., Wilke J.B., Seelbach A., Doeren J., Hindermann M., Butt U.J., Steixner-Kumar A.A., Spieth L., Ronnenberg A., Pan H., Berghoff S.A., Hollmann M., Lühder F., Nave K.A., Bechter K, & Ehrenreich H. (2021). NMDAR1 autoantibodies amplify behavioral phenotypes of genetic white matter inflammation: a mild encephalitis model with neuropsychiatric relevance. Molecular Psychiatry, 27(12), 4974-4983.

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Nanoparticle-mediated Cancer Immunotherapy Protocol

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DOTAP (1,2-dioleoyl-3-trimethylammonium-propane) (890890) and cholesterol (700000) lipids were purchased from Avanti Polar Lipids, Inc., Alabaster, AL, USA. Nanoplasmid (NTC 9385R-MCS) containing the CRT gene was purchased from Aldevron (Lincoln, NE, USA). Chloroform (C2432) was purchased from Sigma-Aldrich, St. Louis, MO, USA. HEPES, sterile 1 M buffer (J848), DMEM (11965092), Fetal bovine serum, FBS (10082147), Penicillin-Streptomycin - PenStrep (15140122), PBS (10010023), Collagenase IV (17104019), were purchased from ThermoFisher/Gibco, Waltham, MA, USA. The following Fluorochrome-conjugated monoclonal antibodies (mAbs) for flow cytometry were purchased from BioLegend, San Diego, CA, USA and were listed: APC-Cy7 anti-CD45 (103115), PerCP anti-CD3 epsilon (100325), PE-Cy5 anti-CD4 (100410), PE-Cy7 anti-CD8a (100721), PE anti-Granzyme B (372208), BV510 anti-INFg (505841), BV510 anti-Ly-6G (127633), PE anti-Ly-6C (128007), BV650 anti-CD11b (101239), APC anti-CD206 (141707), BV421 anti-CD11c (117329). AF488 anti-iNOS (53-5920-82) was procured from eBiosciences/ThermoFisher, Waltham, MA, USA. Liberase (5401119001) was procured from Life Technologies, NY, USA. Transcription factor buffer set (562574) was purchased from BD Biosciences, San Jose, CA, USA. B16F10 was provided by Dr. Mary Jo Turk, Dartmouth. Aged C57BL/6 mice were obtained from NIA, Bethesda, MD, USA.

Singh A., Ashar H., Butcher J.T, & Ranjan A. (2023). Age-associated changes in the gut microbiome impact efficacy of tumor immunomodulatory treatments. Experimental gerontology, 181, 112268.

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