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Cd45.1 pe

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CD45.1-PE is a fluorescently labeled antibody that recognizes the CD45.1 cell surface antigen. It is commonly used in flow cytometry applications to identify and study specific cell populations.

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Lab products found in correlation

Cd45.2 fitc, by Thermo Fisher Scientific (4 mentions) Cd45.2 apc, by Thermo Fisher Scientific (3 mentions) B220 apc cy7, by Thermo Fisher Scientific (3 mentions) Tcrβ pe, by Thermo Fisher Scientific (3 mentions) Cd103 apc, by Thermo Fisher Scientific (3 mentions) F4 80 percp cy5, by BioLegend (2 mentions) Cd11b bv650, by BioLegend (2 mentions) Pd l1 pe, by BioLegend (2 mentions) Cd4 bv421, by BioLegend (2 mentions) Pd1 bv605, by BD (2 mentions) Cd90.1 fitc, by Thermo Fisher Scientific (2 mentions) Cd45.1 apc, by Thermo Fisher Scientific (2 mentions) Cd45.2 alexa700, by Thermo Fisher Scientific (2 mentions) Cd8α percp cy5, by Thermo Fisher Scientific (2 mentions) Cd24 apc cy7, by Thermo Fisher Scientific (2 mentions) Cd44 apcy7, by Thermo Fisher Scientific (2 mentions) Cd90.2 alexa 700, by BioLegend (2 mentions) Mhcii bv421, by BioLegend (2 mentions) Cd8α pe texas red, by Thermo Fisher Scientific (2 mentions) Ly6c bv711, by BioLegend (2 mentions)

10 protocols using cd45.1 pe

1

Multicolor Flow Cytometry Analysis

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Cocultured cells were analyzed by staining with CD45.1 PE, CD45.2 PE, c-Kit APC (eBioscience, San Diego, CA), Sca-1 PE-Cy7, streptavidin APC-Cy7 (BD Pharmingen) and lineage cocktail biotin (a component of StemSep, STEMCELL Technologies). Labeled cells were analyzed using an LSRII flow cytometer (BD Biosciences).
Jeong S.Y., Lyu J., Kim J.A, & Oh I.H. (2020). Ryk modulates the niche activity of mesenchymal stromal cells by fine-tuning canonical Wnt signaling. Experimental & Molecular Medicine, 52(7), 1140-1151.
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2

Multiparametric Flow Cytometry Analysis

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Flow-cytometry antibodies included CD3e-PE, PD1-BV605, CD45-BV786 (BD-Biosciences), CD90.1-FITC, CD62L-PECy7, CD45.1-APC, CD45.1-PE (eBioscience), CD45.2-Alexa700, CD-8α-Percp-Cy5.5, CD4-FITC, CD103-APC, CD24-APCcy7 (Invitrogen), CD44-APCy7, CD4-BV421, F4/80-PercpCy5.5, CD39-PECy7, PDL1-PE, CD90.2-Alexa 700, MHCII-BV421, CD11b-BV650, Ly6C-BV711 (Biolegend), CD8α-PE-Texas-red (Life Technologies, Carlsbad, CA, USA). Blocking PD-1 antibody (clone RMP1-14, BioXCell, Branford, CT, USA) was administered systemically by intraperitoneal injections of 250 μg [41 (link)]. Blocking anti-CD40L (clone MR1, BioXCell) was administered systemically by intraperitoneal injections of 250 μg [19 (link)].
Sharon S., Baird J.R., Bambina S., Kramer G., Blair T.C., Jensen S.M., Leidner R.S., Bell R.B., Casap N., Crittenden M.R, & Gough M.J. (2021). A platform for locoregional T-cell immunotherapy to control HNSCC recurrence following tumor resection. Oncotarget, 12(13), 1201-1213.
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3

Comprehensive Murine and Human Immune Profiling

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Flow cytometry antibodies for murine samples included CD3e-PE, PD1-BV605, CD45-BV786 (BDbiosciences), CD90.1-FITC, CD62L-PECy7, CD45.1-PE (ebioscience), CD45.2-Alexa700, CD-8α-Percp Cy5.5, CD4-FITC, CD103-APC, CD24-APCcy7 (Invitrogen), CD44-APCy7, CD4-BV421, F4/80-PercpCy5.5, CD39-PECy7, PDL1-PE, CD90.2-Alexa 700, MHCII-BV421, CD11b-BV650, Ly6C-BV711 (Biolegend), CD8α-PE-Texas red (life technologies), CD278 (ICOS)-PE (Biolegend). Flow cytometry antibodies for human samples included CD45-BV510, CD3-Alexa700, CD4-BV785, CD8-BV711, CTLA-4-PE/Dazzle 594, 4-1BB-PE (Biolegend), CD39-BV650, PD-1-PE-Cy7 and Ki-67 Alexa 488 (BD Biosciences), CD103-APC and FOXP3-Alexa 700 (eBioscience).
Mouse antibodies for the explant assays included anti-PD1 (RMP1-14) and anti-CTLA-4 (9D9) by BioXCell and OX40 (OX86) kindly provided by Dr. Andrew Weinberg (EACRI). Human antibodies for the explant assays included anti-PD1 and anti-CTLA4 purchased from Invitrogen and anti-OX40 kindly provided by Dr. Andrew Weinberg (EACRI).
Sharon S., Duhen T., Bambina S., Baird J., Leidner R., Bell B., Casap N., Crittenden M., Vasudevan S., Jubran M., Kravchenko-Balasha N, & Gough M. (2021). Explant Modeling of the Immune Environment of Head and Neck Cancer. Frontiers in Oncology, 11, 611365.
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4

Multi-lineage Hematopoietic Cell Analysis

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Total white blood cells from hematopoietic organs were stained in PBS supplemented with 2% FBS with fluorochrome-conjugated mouse antibodies against specific hematopoietic lineage markers. For the analysis of transplant-receiving mice, WBM was stained with fluorochrome-conjugated mouse antibodies to discriminate cells derived from competitors and donors (CD45.1+ and CD45.2+ respectively) and GFP expression was used to precisely define donor-derived cells (GFP+ CD45.2+). Fluorochrome-conjugated mouse antibodies were obtained from Becton Dickinson (Streptavidin/PeCy5, CD117/PeCy7, CD45.1/PE, CD4/PE or PB, CD8/PeCy7 or APC, CD19/APC, B220/APCCy7, TCRβ/PE, CD279/BV421, CD11b/PerCP-Cy5.5, Annexin V/APC, Ki67/PE) and eBiosciences (CD117/APCCy7, Sca-1/APC, CD45.2/APC, Gr1/PE). Hoescht 33342 was obtained from Invitrogen. APC BrdU Flow kit (Becton Dickinson) was used according to the manufacturer’s instruction. Cell sorting was performed either on a MoFlow (Beckman Coulter) or Influx (Becton Dickinson) cell sorter and analysis on a Canto II (Becton Dickinson). FACS data were analyzed by FlowJo Software (v8.8.7).
Scourzic L., Couronné L., Pedersen M.T., Della Valle V., Diop M., Mylonas E., Calvo J., Mouly E., Lopez C.K., Martin N., Fontenay M., Bender A., Guibert S., Dubreuil P., Dessen P., Droin N., Pflumio F., Weber M., Gaulard P., Helin K., Mercher T, & Bernard O.A. (2016). DNMT3AR882H mutant and Tet2 inactivation cooperate in the deregulation of DNA methylation control to induce lymphoid malignancies in mice. Leukemia, 30(6), 1388-1398.
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5

Multi-lineage Hematopoietic Cell Analysis

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Total white blood cells from hematopoietic organs were stained in PBS supplemented with 2% FBS with fluorochrome-conjugated mouse antibodies against specific hematopoietic lineage markers. For the analysis of transplant-receiving mice, WBM was stained with fluorochrome-conjugated mouse antibodies to discriminate cells derived from competitors and donors (CD45.1+ and CD45.2+ respectively) and GFP expression was used to precisely define donor-derived cells (GFP+ CD45.2+). Fluorochrome-conjugated mouse antibodies were obtained from Becton Dickinson (Streptavidin/PeCy5, CD117/PeCy7, CD45.1/PE, CD4/PE or PB, CD8/PeCy7 or APC, CD19/APC, B220/APCCy7, TCRβ/PE, CD279/BV421, CD11b/PerCP-Cy5.5, Annexin V/APC, Ki67/PE) and eBiosciences (CD117/APCCy7, Sca-1/APC, CD45.2/APC, Gr1/PE). Hoescht 33342 was obtained from Invitrogen. APC BrdU Flow kit (Becton Dickinson) was used according to the manufacturer’s instruction. Cell sorting was performed either on a MoFlow (Beckman Coulter) or Influx (Becton Dickinson) cell sorter and analysis on a Canto II (Becton Dickinson). FACS data were analyzed by FlowJo Software (v8.8.7).
Scourzic L., Couronné L., Pedersen M.T., Della Valle V., Diop M., Mylonas E., Calvo J., Mouly E., Lopez C.K., Martin N., Fontenay M., Bender A., Guibert S., Dubreuil P., Dessen P., Droin N., Pflumio F., Weber M., Gaulard P., Helin K., Mercher T, & Bernard O.A. (2016). DNMT3AR882H mutant and Tet2 inactivation cooperate in the deregulation of DNA methylation control to induce lymphoid malignancies in mice. Leukemia, 30(6), 1388-1398.
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6

Multicolor Flow Cytometry Antibody Panel

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Commercial antibodies and staining reagents, from BioLegend: GL7-PacBlue, GL7-PE, GL7-A647, B220-PerCP-Cy5.5, B220-APC-Cy7, IgMb-FITC, IgMa-PE, CD45.2-FITC, CD45.2-APC, CD45.1-FITC, CD45.1-PE, IgD-PB, CD21/35 (7E9)-PE, CD138-PE, CD38-PE-Cy7, CD31-A647, CD157-PE, Streptavidin-PE/Cy7; from eBioscience: CD95 (APO-1/Fas)-PE (clone 15A7) and viability dye Fixable live/dead stain Efluor780; from ThermoFisher Scientific: Rabbit-anti-Goat-A488, Hoechst 33342, DAPI (4',6-Diamidino-2-Phenylindole, Dihydrochloride), Phalloidin-A568; from Southern Biotech: AP-Goat-anti-Mouse IgG, AP-Goat-anti-Mouse IgG2a, AP-Goat-anti-Mouse IgG2c; from Rockland Immunochemicals: Rabbit polyclonal anti-B-Phycoerythrin; from DAKO: Rabbit polyconal anti-Mouse Immunoglobulins-biotin; from Perkin-Elmer: Europium-labeled streptavidin. In-house generated anti-idiotypic Ab, clone 9D11, conjugated to Alexa Fluor 647 or Alexa Fluor 568; in-house generated Rabbit polyclonal anti-C3b conjugated to Alexa Fluor 633.
Degn S.E., van der Poel C.E., Firl D.J., Ayoglu B., Al Qureshah F.A., Bajic G., Mesin L., Reynaud C.A., Weill J.C., Utz P.J., Victora G.D, & Carroll M.C. (2017). Clonal Evolution of Autoreactive Germinal Centers. Cell, 170(5), 913-926.e19.
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7

Multicolor Flow Cytometry Analysis

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For analysis of cell surface markers, cells were stained in PBS containing 2% heat-inactivated FBS with 2 mM EDTA supplementation, unless noted otherwise. Antibodies used were as follows: CD4-PerCP/Cy5.5, CD8α-APC, and CD127-APC (BioLegend); CD4-PE and CD44-PE (BD); CD62L-FITC, CD25-PE, CD45.2-FITC, and CD45.1-PE (eBioscience). Foxp3-APC and eFluor780 (eBioscience), Annexin V-FITC, and 7-AAD (BD) were stained according to the manufacturer’s instructions. Cells were analyzed with FACSCalibur or FACSCanto II flow cytometry (BD) using FlowJo software (Tree Star).
Matsushita M., Freigang S., Schneider C., Conrad M., Bornkamm G.W, & Kopf M. (2015). T cell lipid peroxidation induces ferroptosis and prevents immunity to infection. The Journal of Experimental Medicine, 212(4), 555-568.
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8

Multiparameter Flow Cytometry Staining

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Samples were stained with a 2.4G2 blocking antibody and monoclonal antibodies in 1% PBS-serum for 20 minutes on ice in the dark. For samples stained with CD1 tetramer, cells were incubated for 15 minutes at room temperature in the dark, followed by 15 minutes on ice in the dark. Samples requiring intracellular staining were then fixed and permeabilized using CytoFix/CytoPerm (BD Biosciences) and stained with intracellular antibodies for 15 minutes on ice in the dark. Events were collected on either a FACSAria (BD Biosciences) or a MACSQuant (Miltenyi Biotec). Data were analyzed using FlowJo (Tree Star Inc.). IL-17A-AlexaFluor488, CD45.2-FITC, HSA-FITC, TCRβ-FITC, TCRVβ2-FITC, CD45.1-PE, PLZF-PE, TCRβ-PE, Foxp3-PE-Cy5.5, NK1.1-PerCPCy5.5, TCRβ-PErCPCy5.5, RORγt-PerCPeFluor710, TCRβ8.1/2-PerCPeFluor710, Ly49G2-PerCPeFluor710, IFN-γ-PE-Cy7, NK1.1-PE-Cy7, T-Bet-PE-Cy7, CD45.1-APC, CD25-APC, IL-4-APC, CD4-APCeFluor780, CD44-APCeFluor780, CD45.2-APCeFluor780, CD90.1-APCeFluor780, CD8α-eFluor450, CD3-eFluor450, Ki67-eFluor450, and TNF-α-eFluor540 were purchased from eBioscience (San Diego, CA). Ly49C/I-FITC, Ly49A/D-PE, and CD4-PerCP were purchased from BD Pharmingen. TCRVβ7-PE was purchased from Biolegend. CD1d tetramer loaded with α-GalCer, CD1d tetramer loaded with PBS-57 as well as unloaded controls were provided by the NIH Tetramer Facility.
Anderson C.K., Salter A.I., Toussaint L.E., Reilly E.C., Fugère C., Srivastava N., Kerr W.G, & Brossay L. (2015). Role of SHIP1 in iNKT cell development and functions. Journal of immunology (Baltimore, Md. : 1950), 195(5), 2149-2156.
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9

Hematopoietic Stem Cell Transplantation

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Cell preparations were injected intravenously into irradiated (9.5 Gy) recipients that differed in their CD45 isoform expression. For the primary transplants, E11 AGM, E12 Plac, E12 FL, and E14 FL were injected at 1, 0.5, 0.5, and 0.004 embryo equivalent per recipient, respectively, together with 100,000 total BM helper cells. For the secondary and tertiary transplants, between two and three million total BM cells (without helper cells) were injected, which were normalized according to the level of donor chimerism. At the indicated time points, donor contribution to the peripheral blood was analyzed by flow cytometry using CD45.1-PE (eBioscience A20), CD45.2-FITC (eBioscience 104), CD11b-eFluor450 (eBioscience M1/70), Gr1-AF700 (BioLegend RB6-8C5), CD19-BV605 (BioLegend 1D2), B220-PECy7 (eBioscience RA3-6B2), CD3-APC (BioLegend I45-2C11), and ckit-APCeFluor780 (eBioscience 2B8). Mice were considered positive for repopulation if the donor contribution to the total peripheral blood was >5%. Peripheral blood counts were performed on an ABC blood counter (Woodley).
Barrett N.A., Malouf C., Kapeni C., Bacon W.A., Giotopoulos G., Jacobsen S.E., Huntly B.J, & Ottersbach K. (2016). Mll-AF4 Confers Enhanced Self-Renewal and Lymphoid Potential during a Restricted Window in Development. Cell Reports, 16(4), 1039-1054.
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10

Multiparametric Immune Cell Profiling

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The following antibodies were used to identify cell subsets: Ly6C-FITC (AL-21, BD Biosciences), F4/80-APC eFluor660 (BM8, eBioscience), Siglec-F-PE or AR700 (E50-2440, BD Biosciences), CD11c-PE or BV711 (HL3, BioLegend), Ly6G-PerCP eFluor710 or V450 (1A8, eBioscience or BD Biosciences), CD45.2-APC/Fire750 or V450 (104, BioLegend or eBioscience), CD45.1-PE (A20, eBioscience), and CD11c-BV711 (N418, BioLegend). Dead cells were excluded from analyses using Fixable Viability Dye APC BV506 (eBioscience).
Surface staining: Cells were treated with anti-CD16/CD32 Fc receptor blocking antibody (clone 2.4G2) in 1x PBS (1% FBS) for 10 minutes on ice. Cells were then centrifuged and resuspended in 1x PBS (1% FBS) containing antibodies and incubated for 15 minutes on ice. Cells were washed with 1x PBS then incubated with Fixable Viability Dye diluted in 1x PBS for 15 minutes on ice.
Cells were washed, then fixed with 1% paraformaldehyde for 15 minutes on ice. Samples were acquired using an Attune NxT Acoustic Focusing Cytometer with Attune Software.
Analyses were performed using FlowJo v10 software (Tree Star, Inc.). Gate placement was determined using fluorescence minus one and unstained control samples.
Crane M.J., Xu Y., Monaghan S.F., Hall B.M., Albina J.E., Henry W.P., Tran H.L., Chhabria K., Jordon A.R.D., Carlsen L., & Jamieson A.M. (2020). Pulmonary infection interrupts acute cutaneous wound healing through disruption of chemokine signals.
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