Anti cd45.1 a20

Manufactured by Thermo Fisher Scientific

Sourced in United States

The Anti-CD45.1 (A20) is a laboratory reagent used for the identification and quantification of CD45.1-positive cells in various research applications. It is a monoclonal antibody that specifically binds to the CD45.1 antigen, which is expressed on the surface of certain leukocyte subpopulations. This product can be used in flow cytometry, immunohistochemistry, and other techniques to study the phenotype and distribution of CD45.1-positive cells.

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Close spelling variants of this product

Anti-CD45 (119 citations) CD45.1 (A20, (22 citations) Anti-mouse CD45.1 PE-Cy7 (A20, (3 citations) Biotinylated anti-CD45.1 (clone A20; (3 citations) Anti-CD45.1 (clone A20; (2 citations)

7 protocols using anti cd45.1 a20

Murine T Cell Activation Assay

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Tamoxifen was purchased from Sigma-Aldrich (St. Louis, MO). Carboxyfluorescein succinimidyl ester (CFSE) and LIVE/DEAD Fixable Dead Cell stain was purchased from Molecular Probes, Invitrogen (Carlsbad, CA). Antibodies were purchased from either BD Pharmingen (San Diego, CA): anti-CD4 (RM4-5), anti-CD25 (PC61), anti-CD3 (2C11), and anti-CD28 (37.51); Biolegend (San Diego, CA): anti-CD45.2 (104), anti-CD45.1 (A20), and anti-CD4 (GK1.5), or eBioscience (San Diego, CA): anti-Foxp3 (FJK-16s), anti-CD69 (H1.2F3), anti-glucocorticoid-induced TNFR family related gene (GITR) (DTA-1), and anti-CTLA-4 (UC10-4B9).

Schmidt A.M., Lu W., Sindhava V.J., Huang Y., Burkhardt J.K., Yang E., Riese M.J., Maltzman J.S., Jordan M.S, & Kambayashi T. (2015). Regulatory T cells require TCR signaling for their suppressive function. Journal of immunology (Baltimore, Md. : 1950), 194(9), 4362-4370.

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Comprehensive Immunostaining Panel for Immune Cell Analysis

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Anti-Nono (Cat# 11058-1-AP and 66361-1-Ig) was from Proteintech. Anti-Elk3 (Cat# NBP1-83960) was from Novus Biologicals. Anti-Tmem241 (Cat# 203644-T32) was from Sinobiological. Anti-Lineage cocktail (Cat# 88-7772-72), Anti-CD127 (A7R34), anti-Sca-1 (D7), anti-Flt3 (A2F10), anti-α₄β₇ (DATK32), anti-Id2 (ILCID2), anti-PLZF (Mags.21F7), anti-Eomes (Dan11mag), anti-NKp46 (29A1.4), anti-NK1.1 (PK136), anti-CD45 (30-F11), anti-CD25 (PC61.5), anti-Gata3 (TWAJ), anti-RORγt (AFKJS-9), anti-Bcl11b (1F8H9), anti-CD3 (17A2), anti-CD19 (1D3), anti-KLRG1 (2F1), anti-CD90 (HIS51), anti-IL-22 (IL22JOP), anti-Ki67 (SolA15), anti-CD45.2 (104), anti-BrdU (BU20A), anti-PD-1 (J43) and anti-CD45.1 (A20) were purchased from eBiosciences (San Diego, USA). Anti-c-Kit (2B8), anti-CD150 (TC15-12F12.2), anti-CD48 (HM48-1), and anti-CD49a (HMα1) were purchased from Biolegend (California, USA). Anti-β-actin (Cat# RM2001) was purchased from Beijing Ray Antibody Biotech. Paraformaldehyde (PFA) and 4’,6-diamidino-2-phenylindole (DAPI) were from Sigma. IL-22 ELISA kit was purchased from Neobioscience.

Liu N., He J., Fan D., Gu Y., Wang J., Li H., Zhu X., Du Y., Tian Y., Liu B, & Fan Z. (2022). Circular RNA circTmem241 drives group III innate lymphoid cell differentiation via initiation of Elk3 transcription. Nature Communications, 13, 4711.

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Immunofluorescence Analysis of Murine Ear Tissue

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Ear pinnae were split with forceps, fixed in 1% paraformaldehyde in PBS (Electron Microscopy Sciences) overnight at 4°C and blocked in 1% BSA + 0.25% Triton X in PBS for 2 hours at room temperature. Tissues were first stained with anti-CD4 (RM4–5, eBioscience), anti-CD8α (clone 53–6.7, eBioscience), anti-CD45.1 (A20, eBioscience), anti-CD49f (GoH3, eBioscience) and/or anti-GFP (A21311, Life Technologies) antibodies overnight at 4°C, washed three time with PBS and then stained with 4,6-diamidino-2-phenylindole (DAPI, Sigma-Aldrich) overnight before being mounted with ProLong Gold (Molecular Probes) antifade reagent. Ear pinnae images were captured on a Leica TCS SP8 confocal microscope equipped with HyD and PMT detectors and a 40× oil objective (HC PL APO 40X/1.3 oil). Images were analyzed using Imaris software (Bitplane).

Harrison O.J., Linehan J.L., Shih H.Y., Bouladoux N., Han S.J., Smelkinson M., Sen S.K., Byrd A.L., Enamorado M., Yao C., Tamoutounour S., Van Laethem F., Hurabielle C., Collins N., Paun A., Salcedo R., O’Shea J.J, & Belkaid Y. (2018). Commensal-specific T cell plasticity promotes rapid tissue adaptation to injury. Science (New York, N.Y.), 363(6422), eaat6280.

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Detailed Immunological Assay Protocol

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The clone numbers for the antibodies used in these experiments: anti- F. tularensis lipopolysaccharide (1.B.288, US Biologicals; Salem, MA), anti-MHC I H2-Kd (SF1-1.1.1, eBioscience; San Diego, CA), anti-MHC I H2-Kb (AF6-88.5.5.3, eBioscience), anti-MHC I HLA-A2 (BB7.2, eBioscience), anti-CD45.1 (A20, eBioscience), anti-CD45 (30-F11, eBioscience), anti-MHC I H2-Kb-SIINFEKL (25-D1.16, eBioscience).
The catalog number and company for critical reagents used in these experiments: Cell Trace Red DDAO-SE (C34553, Life Technologies; Carlsbad, CA), Calcein-AM (C3099, Life Technologies), Soy Lecithin (Cas number 8002-43-5, Acros; Waltman, MA), phalloidin (A22287, Life Technologies), 3 um pore Transwells (3402 Costar; Corning, NY), gentamicin (15750-060, Gibco; Carlsbad, CA)
The beads (M-1002-010, Solulink; San Diego, CA) used in these experiments were labeled with AF488 succinimidyl ester (A-20100, Life Technologies) to make fluorescent beads.

Steele S., Radlinski L., Taft-Benz S., Brunton J, & Kawula T.H. (2016). Trogocytosis-associated cell to cell spread of intracellular bacterial pathogens. eLife, 5, e10625.

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In Vivo Cytotoxicity Assay with CFSE-Labeled Cells

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LCMV-infected (weeks 4–6) or naive mice received CFSE⁺CD45.1⁺ splenocytes pulsed with gp33 peptide and a nonpulsed CFSE⁺CD45.2⁺ reference population. 5 µM CFSE was used to label the cell populations. Cells were pulsed for 60 min with 5 µM gp33 peptide. 2 h after the transfer of the target cells, total splenocytes were harvested and stained with anti-CD45.1 (A20; eBioscience) and self-conjugated anti-CD45.2 (custom purified from hybridoma 104; Bio X Cell) and analyzed by flow cytometry. Using the no-peptide reference population, the fraction of eliminated peptide-containing target cells was calculated.

Utzschneider D.T., Alfei F., Roelli P., Barras D., Chennupati V., Darbre S., Delorenzi M., Pinschewer D.D, & Zehn D. (2016). High antigen levels induce an exhausted phenotype in a chronic infection without impairing T cell expansion and survival. The Journal of Experimental Medicine, 213(9), 1819-1834.

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Intestinal Epithelial Cell Phenotyping

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Colon cells were stained with Fc Block (Clone 2.4G2) followed by staining with fluorescently labelled antibodies in IEC buffer (PBS with 5% FBS and 2 mM EDTA to reduce cell clumping) for IECs or 2% FBS in PBS for T cells on ice in 1.5 ml microcentrifuge tubes. For intracellular staining, cells were fixed and permeabilized using BD Cytofix/Cytoperm kit (BD Bioscience). Samples were acquired on an Attune NxT flow cytometer (Life Technologies) and analysed with FlowJo software. Cells were sorted on either a BD ARIA II or S6 Enceladus (BD Biosciences). The following antibodies/reagents were used: anti-CD45 (30-F11; Biolegend), anti-EPCAM1 (G8.8; ThermoFisher), anti-FABP2 (polyclonal; R&D/Fisher), anti-goat IgG (ThermoFisher), anti-LY6G (1A8; BioLegend), anti-MHCII (M5/114.15.2; Fisher), Live/Dead Fixable Near-IR dead cell dye (ThermoFisher), anti-CD4 (RM4-5; BioLegend), anti-TCRβ (Η57−597; ThermoFisher), anti-CD44 (IM7; BioLegend), anti-CD45.1 (A20; ThermoFisher), anti-CD45.2 (104; BD Biosciences), anti-IL-17A (TC11-18H10; BD Biosciences), anti-IFNγ (XMG1.2; ThermoFisher), and anti-IL-22 (1H8PWSR; ThermoFisher).

Zindl C.L., Wilson C.G., Chadha A.S., Duck L.W., Cai B., Harbour S.N., Nagaoka-Kamata Y., Hatton R.D., Gao M., Figge D.A, & Weaver C.T. (2024). Distal colonocytes targeted by C. rodentium recruit T-cell help for barrier defence. Nature, 629(8012), 669-678.

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Comprehensive Antibody Panel for Murine Cell Analysis

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Antibodies used in the experiments with mouse cells were obtained from Biolegend unless otherwise stated: anti-CD3 (17A2); anti-CD4 (GK1.5); anti-CD8 (53-6.7); anti-B220 (RA3-6B2); anti-CD25 (PC61.5); anti-CD44 (IM7); anti-CD45 (30-F11); anti-CD62L (MEL-14); anti-CD45.1(A20); anti-CD45.2 (104); anti-NKG2D (CX5); anti-PD-1 (29F.1A12); anti-TIM-3 (RMT3-23); anti-CD200 (OX-90); anti-EpCAM (G8.8); anti-MHC I (36-7-5); anti-MHC II (M5/114.15.2); anti-LAG-3 (C9B7W; Thermo Fisher Scientific); anti-pan NK cells (DX5; Thermo fisher); anti-Foxp3 (FJK-16s; Thermofisher); IL-17 (TC11-18H10.1); anti-TNF-α (TN3-19.12); anti-IFN-γ (XMG1.2); anti-Eomes (Dan11mag; Thermo fisher); anti-TOX (REA473; Miltenyi Biotec), anti-pStat1 (Tyr701) (58D6; Cell Signaling); anti-pStat5 (Tyr694) (C71E5; Cell Signaling); anti-Actin (SCBT). The following mAbs raised against human antigens were all purchased from Biolegend: CD3 (OKT3); CD4 (OKT4); CD8 (SK1); NKG2D (1D11); IL-17 (BL168); IFN-γ (B27). Viable cell populations were gated based on forward and side scatters and by fixable blue (Thermo Fisher Scientific) staining. Samples were collected on an LSR II and analyzed with FlowJo software (Tree Star).

Dai Z., Sezin T., Chang Y., Lee E.Y., Wang E.H, & Christiano A.M. (2022). Induction of T cell exhaustion by JAK1/3 inhibition in the treatment of alopecia areata. Frontiers in Immunology, 13, 955038.

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