Anti cd45.2 pe clone 104

Manufactured by Thermo Fisher Scientific

Anti-CD45.2-PE (clone 104) is a fluorescently-labeled antibody product that binds to the CD45.2 cell surface antigen. It can be used for the identification and analysis of cells expressing the CD45.2 marker in flow cytometry applications.

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Lab products found in correlation

Facscalibur, by BD (2 mentions) Anti cd11c apc clone n418, by Thermo Fisher Scientific (1 mentions) 7 aad, by Thermo Fisher Scientific (1 mentions) Pharm lyse, by BD (1 mentions) Anti il 17 fitc, by Thermo Fisher Scientific (1 mentions) Anti gr 1 apc, by BioLegend (1 mentions) Anti cd11b fitc, by BioLegend (1 mentions) Facscanto flow cytometer, by BD (1 mentions)

Close spelling variants of this product

Anti-CD45.2-PE (clone 104) monoclonal antibodies Anti-CD45.2 (clone 104, CD45.2 (clone 104, Anti-CD45.2 (104) CD45.2 (104, CD45.2-PE Anti-CD45-PE CD45-PE Anti-CD45

3 protocols using anti cd45.2 pe clone 104

Multicolor Flow Cytometry of Lung Immune Cells

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The pellet of BALF cells was resuspended in 300 µL of PBS with bovine serum albumin (BSA) and used for flow cytometry analysis. The cells were pipetted through cell strainer cap directly to the well of 96-well plate and were centrifuged (5 min, 1500 rpm, 4℃). Then supernatant was discarded and cells were resuspended in 54 µL of 5% normal rat plasma in PBS with BSA and blocked for 15 minutes on ice. Then, the cells were divided into two parts and to each 3 µL of antibody cocktail was added (according to Table 2). The cells were stained on ice for 30 minutes in dark, washed twice in PBS with BSA and resuspended in PBS with BSA and NaN₃ for the flow cytometry analysis (Guava, Merck).Table 2

Antibodies Used for Flow Cytometry Analysis

Antibody	Cat. No.
Anti-CD11b-AF488 (clone M1/70)	Biolegend, 101217
Anti-CD45.2-PE (clone 104)	eBiosciences, 12–0454
Anti-Ly6G-PerCP-Cy5.5 (clone 1A8)	Biolegend, 127616
Anti-CD11c-APC (clone N418)	eBiosciences, 17–0114–82

Dymek B., Sklepkiewicz P., Mlacki M., Güner N.C., Nejman-Gryz P., Drzewicka K., Przysucha N., Rymaszewska A., Paplinska-Goryca M., Zagozdzon A., Proboszcz M., Krzemiński Ł., von der Thüsen J.H., Górska K., Dzwonek K., Zasłona Z., Dobrzanski P, & Krenke R. (2022). Pharmacological Inhibition of Chitotriosidase (CHIT1) as a Novel Therapeutic Approach for Sarcoidosis. Journal of Inflammation Research, 15, 5621-5634.

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Multilineage Hematopoietic Reconstitution Assay

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Cell suspensions from embryos at different stages were injected into irradiated adult recipients (CD45.1/2) either directly (E13.5 FL, 0.2 embryo equivalents per recipient mouse) or after explant culture (E11.5 AGM, 0.1 embryo equivalents per recipient mouse), along with 80,000 CD45.1/1 bone marrow carrier cells. Recipients were irradiated by split dose (472 + 472 rad with 3-hr interval) of γ irradiation. Donor-derived chimerism was monitored in blood at 6 and 16 weeks after transplantation using FACScalibur (BD). Peripheral blood was collected by bleeding the lateral tail vein into 500 μL 5 mM EDTA/PBS, and erythrocytes were depleted using PharM Lyse (BD). Cells were stained with anti-CD16/32 (Fc-block, cat. no. 16-0161-86), CD45.1-APC (clone A20, cat. no. 17-0453-82), and anti-CD45.2-PE (clone 104, cat. no. 12-0454-83) monoclonal antibodies (eBioscience). Appropriate isotype controls were used. Dead cells were excluded using 7AAD (eBioscience). Mice demonstrating ≥5% donor-derived multilineage chimerism after 8 weeks were considered to be reconstituted.

Senserrich J., Batsivari A., Rybtsov S., Gordon-Keylock S., Souilhol C., Buchholz F., Hills D., Zhao S, & Medvinsky A. (2018). Analysis of Runx1 Using Induced Gene Ablation Reveals Its Essential Role in Pre-liver HSC Development and Limitations of an In Vivo Approach. Stem Cell Reports, 11(3), 784-794.

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Intravascular Leukocyte Discrimination Protocol

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Intravascular staining for the discrimination of vascular and lung tissue leukocytes was conducted as previously described [26 (link)]. Anti-CD45.2 PE (clone 104, catalog number 12-0454-81; eBioscience, San Diego, California) was diluted to 25 mg/mL in sterile PBS; 100 μL of the solution was intravenously injected via the mouse tail 3 minutes before sacrificing the mice. The stained CD45.2⁺ cells are blood-borne in the mouse lungs and can be discriminated from cells localized in the lung tissue. Cellular staining and flow cytometry analyses for T helper (Th) cells and neutrophils were conducted as previously described [27 (link)]. In brief, cells were stained for surface markers with anti-CD4 fluorescein isothiocyanate (FITC; clone: GK1.5; eBioscience) for Th cells, and with anti-CD11b FITC (clone: M1/70; Biolegend, San Diego, California) and anti-Gr-1 APC (clone: RB6-8C5; Biolegend) for neutrophils. For intracellular staining, fixed cells were permeabilized and stained with anti–IL-17 FITC (catalog number 11-7177-81; eBioscience) and anti–interferon gamma (IFN-γ) APC (clone: XMG1.2; eBioscience) for 60 minutes in the dark at 4°C. Cells were washed with 1 mL of permeabilization buffer and resuspended in staining buffer for analysis. Samples were analyzed using a FACSCalibur or FACSCanto flow cytometer (BD Biosciences) and the FlowJo software (Tree Star, Ashland, Oregon).

Fan X., Li N., Xu M., Yang D, & Wang B. (2021). Intrapulmonary Vaccination Induces Long-lasting and Effective Pulmonary Immunity Against Staphylococcus aureus Pneumonia. The Journal of Infectious Diseases, 224(5), 903-913.

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