Cd45.1 pe

Manufactured by BD

CD45.1-PE is a fluorescently labeled antibody that binds to the CD45.1 antigen, which is a common leukocyte antigen expressed on the surface of certain immune cells. The PE (phycoerythrin) fluorescent label allows for the detection and analysis of cells expressing CD45.1 using flow cytometry or other fluorescence-based techniques.

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CD45-PE (130 citations) Anti-CD45.1-PE (7 citations) Anti-CD45.1-PE (clone A20; (2 citations) Anti-mouse CD45.1-PE (2 citations)

5 protocols using cd45.1 pe

Isolation and Sorting of γδ T Cells

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One hundred microliters of antibody staining solution was prepared in PBS containing 0.5% BSA and 2 mM EDTA and added to the isolated cells resuspended in 100 µl staining buffer as described above. Cells were incubated for 20 min on ice, washed three times with 2 ml of 0.5% BSA in PBS and resuspended in 3 ml after the last wash for cell sorting. The following antibodies were used: TCRγδ‐APC (BioLegend, 1:100 dilution), TCRβ‐BV421 (BioLegend, 1:100 dilution), CD45.1-PE (BD Biosciences, 1:100 dilution), CD45.2-AF700 (BioLegend, 1:100 dilution), CD45.2-PerCP5.5 (BD Biosciences, 1:100 dilution), CD160-PECy7 (BioLegend, 1:100 dilution), Ly6C-BV510 (BioLegend, 1:100 dilution) and CD62L-BV421 (BioLegend, 1:100 dilution). Zombie Aqua and Zombie Green fixable viability kits (BioLegend) were used to distinguish dead and living cells. Living TCRγδ⁺ single γδ T cells were sorted in BSA-coated tubes containing 50 µl of PBS using a FACSAria cell sorter (BD Biosciences) equipped with BD FACSDiva software (v8.0.2). Using pulse geometry gates (FSC‐W × FSC‐H and SSC‐W × SSC‐H), doublets/multiplets were excluded. After the completion of sorting, the cells were processed through the different 10x Genomics workflows.

du Halgouet A., Bruder K., Peltokangas N., Darbois A., Obwegs D., Salou M., Thimme R., Hofmann M., Lantz O, & S.a.g.a.r. (2024). Multimodal profiling reveals site-specific adaptation and tissue residency hallmarks of γδ T cells across organs in mice. Nature Immunology, 25(2), 343-356.

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Multiparameter Flow Cytometry of Immune Cells

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For surface staining, cells were washed with PBS and blocked with PBS plus 5% FBS, 2.5% mouse serum and 0.5% anti Fcγ III/II (anti-CD16/32) for 15min. Cells were washed with PBS and incubated with viability dye (1:500, eBioscience) and antibodies (1:200) for 15 min. Cells were washed with PBS and analyzed by LSR II (BD Biosciences). Flow cytometric data were further analyzed with FlowJo software (TreeStar).
For intracellular staining, cells were incubated with Brefeldin A (1:1000, eBioscience) in RPMI media plus 10% FBS in 37 C for 4–5h. Then cells were washed, blocked and stained according to the Intracellular Staining protocol (eBioscience). Cells were washed with PBS and analyzed by BD LSR II.
Antibodies used in this paper include Ly6C-APC Cy7, Ly6C-APC, F4/80-PE, F4/80-e450, Ly6G 1A8-PE Cy7, Ly6G 1A8-FITC (BD), CD45.2-APC, CD45.2-AF700, CD45.1-PE, CD45.1-AF700, CD3-e450, NK1.1-AF780, CD11b-PerCP Cy5.5 (BD), CD206-PE, Arginase1-PE (R & D), Anti-mouse Relm-α antibody (PeproTech). Antibodies were from eBioscience unless stated otherwise.

Xu Y., Huang L., Kirschman J.L., Vanover D.A., Tiwari P.M., Santangelo P.J., Shen X, & Russell D.G. (2018). Exploitation of synthetic mRNA to drive immune effector cell recruitment and functional re-programming in vivo.. Journal of immunology (Baltimore, Md. : 1950), 202(2), 608-617.

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Competitive Hematopoietic Cell Transplantation

Cited 1 time

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Competitive transplantation was performed on a head-to head basis as previously described [10 (link),13 (link),27 (link)]. Five mice per treatment group were used in all experiments to assess the mean percent chimerism at various times after HCT. The proportion of CD45.1, CD45.2, and CD45.1/CD45.2 cells in peripheral blood was determined monthly by flow cytometry. CD45.1-PE and CD45.2-FITC were purchased from BD Biosciences, San Jose, Ca.

Broxmeyer H.E, & Pelus L.M. (2014). Inhibition of DPP4/CD26 and dmPGE2 treatment enhances engraftment of mouse bone marrow hematopoietic stem cells. Blood cells, molecules & diseases, 53(0), 34-38.

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Quantifying Pulmonary Progenitor Cells

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Lung cells were obtained from digesting masticated lungs in a solution of 5% fetal bovine serum, 1 mg/mL collagenase type IV, and 0.02 mg/mL DNase I (Sigma-Aldrich) in RPMI 1640 medium for 45 minutes at 37^oC. Peripheral blood mononucleocytes (PBMCs) were isolated from blood drawn from the right jugular. Red blood cells (RBCs) were lysed using RBC lysis buffer (BioLegend). For quantifying the number of PACs in the lungs and peripheral blood, both PBMCs and lung cells were stained with anti- Ter119-Pacific Blue, CD11b-redFuor (Tonbo Biosciences, catalog #10141–234 and #80–0112), CD31-PECy7 (BioLegend, catalog #102524), and DAPI. To quantify engraftment, PBMCs were stained with anti-Ter119-Pacific Blue, CD45.1-PE (BD Biosciences, catalog #561872), CD45.2-PerCPCy5.5 (Tonbo Biosciences, catalog #65–0454), and DAPI. Flow cytometry was performed using a BD LSRFortessa and the data analyzed using FlowJo. Antibody-conjugated beads (Thermo Fisher) were used for single stain compensation controls. Unstained cells and fluorescence-minus-one controls were used for gating populations in all flow cytometry experiments.

Bloodworth N.C., Clark C.R., West J.D., Snider J.C., Gaskill C., Shay S., Scott C., Bastarache J., Gladson S., Moore C., D’Amico R., Brittain E.L., Tanjore H., Blackwell T.S., Majka S.M, & David Merryman W. (2018). Bone Marrow-Derived Proangiogenic Cells Mediate Pulmonary Arteriole Stiffening via Serotonin 2B Receptor Dependent Mechanism. Circulation research, 123(12), e51-e64.

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Bone Marrow Transplantation in Arhgap21 Mice

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Bone marrow transplantation was performed by injecting normal hematopoietic cells in Arhgap21^+/− or WT mice. Briefly, 10⁶ bone marrow cells from B6.SJL/BoyJ (PEP; CD45.1+, Jackson Labs) mice were transplanted into 9.5Gy sub-lethally irradiated WT (n = 5) or Arhgap21^+/− (n = 8) (CD45.2+) recipient mice. All mice were 10-weeks old when transplanted. Donor reconstitution (CD45.1+) and hematological parameters (hemoglobin, platelets, and WBC) were evaluated every 4 weeks after transplant until 16 weeks post-transplantation, when animals were terminated. PB was collected by ocular cavity and hematological parameters were analyzed using a CELL-DYN Emerald Hematology System counter (Abbott Laboratories). Chimerism was evaluated by FACScalibur flow cytometer using mAb CD45.1-PE (BD Bioscience).

Pissarra M.F., Torello C.O., Gomes R.G., Shiraishi R.N., Santos I., Vieira Ferro K.P., Lopes M.R., Bergamo Favaro P.M., Olalla Saad S.T, & Lazarini M. (2021). Arhgap21 Deficiency Results in Increase of Osteoblastic Lineage Cells in the Murine Bone Marrow Microenvironment. Frontiers in Cell and Developmental Biology, 9, 718560.

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