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Anti cd45.1 a20

Manufactured by BioLegend
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Anti-CD45.1 (A20) is a lab equipment product from BioLegend that detects the CD45.1 allotype. It is a widely used marker for identification and analysis of cells expressing the CD45.1 antigen.

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Lab products found in correlation

Anti cd45.2 104, by BioLegend (4 mentions) Anti cd11c n418, by BioLegend (3 mentions) Anti cd45 30 f11, by BioLegend (3 mentions) Anti nk1.1 pk136, by BioLegend (3 mentions) Anti cd3 17a2, by BioLegend (3 mentions) Anti cd44 im7, by BioLegend (3 mentions) Celltrace violet, by Thermo Fisher Scientific (3 mentions) Anti cd16 32 2.4g2, by BD (2 mentions) Anti il 7rα a7r34, by BioLegend (2 mentions) Anti siglecf e50 2440, by BD (2 mentions) Anti cd11b m1 70, by BioLegend (2 mentions) Anti cd19 6d5, by BioLegend (2 mentions) Anti ly6g 1a8, by BioLegend (2 mentions) Anti ki67 sola15, by Thermo Fisher Scientific (2 mentions) Aurora spectral cytometer, by Tree Star (2 mentions) Anti human vβ5.1 lc4, by Thermo Fisher Scientific (2 mentions) Anti vβ11 kt11, by BioLegend (2 mentions) Anti mouse cd185 cxcr5 l138d7, by BioLegend (2 mentions) Anti mouse cd279 pd 1 29f 1a12, by BioLegend (2 mentions) Zombie red, by BioLegend (2 mentions)

Close spelling variants of this product

Anti-CD45.1-PerCP/Cy5.5 (A20), Anti-CD45.1 FITC (clone A20, Anti-CD45.1 (clone A20, Alexa647 anti-mouse CD45.1 (clone A20), (PerCP)-anti-CD45.1 (A20), Anti-mouse CD45.1 (Clone A20, Biotin-conjugated anti-CD45.1 (clone A20, CD45.1 (A20, Anti-CD45

10 protocols using anti cd45.1 a20

1

Cytofluorimetry Gating and Proliferation

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For cytofluorimetry, gating was performed on viable cells based on forward scatter and side scatter profiles. Fluorescently labeled antibodies were purchased from eBioscience (anti-CD45.1, A20; anti-CD4, RM4–5; anti-CD11c, N418) or BioLegend (anti-CD45.2, 104; anti–I-A/I-E, M5/114.15.2). Proliferation was assessed using the dye CellTrace Violet (Life Technologies) according to manufacturer’s instructions. Staining for cell surface markers was performed at 4°C for 30 m. Flow cytometry data were acquired on an LSRFortessa (Becton Dickinson) instrument and analyzed with the FlowJo software package (Tri-Star).
Cheloha R.W., Woodham A.W., Bousbaine D., Wang T., Liu S., Sidney J., Sette A., Gellman S.H, & Ploegh H.L. (2019). Recognition of MHC-II peptide ligands that contain β-amino acids. Journal of immunology (Baltimore, Md. : 1950), 203(6), 1619-1628.
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2

Immunophenotyping of Murine Splenocytes and Thymocytes

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Mouse splenocytes and thymocytes were prepared from pooled thymus or spleen. Thymic and splenic cells were pre-incubated with Fc-block before staining with other antibodies. The mouse antibodies used included anti-CD4 (RM4–5, eBioscience); anti-CD8 (53–6.7, eBioscience); anti-CD45.1 (A20, BioLegend), anti-CD45.2 (104, eBioscience); anti-CD25 (PC61.5, eBioscience); anti-CD69 (H1.2F3, eBioscience); anti-CD24 (M1/69, eBioscience); anti-Helios (22F6, eBioscience) before flow cytometry analysis. For intracellular staining of Ki-67 (B56, BD), pSTAT5 (47/Stat5(pY694), BD) and Foxp3 (NRRF-30, eBioscience), cells were fixed and permeabilized with BD Cytofix/Cytoperm™ Fixation / Permeabilization Solution Kit (554,714, BD) and stained according to the manufacturer’s protocols. The CD4+ T cells from the thymus and spleen of WT or RelB−/− mice were analyzed by flow cytometry and gated as shown in the Supplementary Figure 1. The samples were analyzed using a BD LSRFortessa flow cytometer and FlowJo software (Tree Star Inc). All single-cell suspensions from the tissues were stained with Abs diluted in PBS containing 2% FCS for 30 min on ice.
Zhou S., Wu W., Wang Z., Wang Z., Su Q., Li X., Yu Y., Zhang W., Zhu M, & Lin W. (2020). RelB regulates the homeostatic proliferation but not the function of Tregs. BMC Immunology, 21, 37.
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3

Multicolor Flow Cytometry Staining

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Anti-CD11c (N418), anti-F4/80 (BM8), anti-CD19 (6D5), anti-CD11b (M1/70), and anti-CD45.1 (A20) were from BioLegend. Anti-ST2 (U29-93), anti-CD3 (17A2), anti-CD4 (RM4-5), anti-IFNγ (XMG1.2), and anti-CD16/32 (2.4G2) were from BD Biosciences. CellTrace Violet, Fixable Viability Dye eFluor 780, and Foxp3 (FJK-16s) were from Thermo Fisher Scientific. Anti-4-1BB (158332) was from R&D.
Lu D.R., Wu H., Driver I., Ingersoll S., Sohn S., Wang S., Li C.M, & Phee H. (2020). Dynamic changes in the regulatory T-cell heterogeneity and function by murine IL-2 mutein. Life Science Alliance, 3(5), e201900520.
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4

Characterization of CD8+ T Cell Phenotypes

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Single-cell suspensions were prepared from the spleen, LNs, and surface or intracellularly stained as described (Shan et al., 2022a (link)). The fluorochrome-conjugated antibodies were as follows: anti-CD8 (53–6.7), anti-TCRβ (H57-597), anti-CD45.2 (104), anti-Granzyme B (GB12), anti-IFN-γ (XMG1.2), anti-Tbet (4B10), anti-CD62L (MEL-14), anti-KLRG1 (2F1), anti-CD25 (PC61.5), anti-CD69 (H1.2F3), and anti-CD44 (IM7) were from Thermo Fisher Scientific; anti-Tcf1 (C63D9) from Cell Signaling Technology; anti-IL-7Rα (A7R34) and anti-CD45.1 (A20) were from BioLegend. For detection of Tcf1 and Tbet proteins, surface-stained cells were fixed and permeabilized with the Foxp3/Transcription Factor Staining Buffer Set (eBiosciences, Thermo Fisher Scientific), followed by incubation with corresponding fluorochrome-conjugated antibodies. Apoptotic cells were detected with FLICA 660 Caspase-3/7 detection kit (Bio-Rad). Data were collected on FACSCelesta or FACSVerse (BD Biosciences) and were analyzed with FlowJo software V10.2 (TreeStar). For validation of CTCF deletion efficiency, cell lysates from sorted GFP+ naive or early effector CD8+ T cells were immunoblotted with anti-CTCF antibody (D31H2; Cell Signaling Technology) following standard protocols.
Liu J., Zhu S., Hu W., Zhao X., Shan Q., Peng W, & Xue H.H. (2023). CTCF mediates CD8+ effector differentiation through dynamic redistribution and genomic reorganization. The Journal of Experimental Medicine, 220(4), e20221288.
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5

Multiparametric Flow Cytometry Profiling

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For cell surface staining, cells were pre-incubated with 2.4G2 Fcγ RII/RIII blocking mAb for 15 minutes and then stained with the appropriate combinations of mAbs diluted in HBSS + 2% FBS for 30 minutes at 4°C. Cells were analyzed on a BD FACS CantoII, or Cytek Aurora Spectral Cytometer and data were processed with FlowJo software (TreeStar). The complete list of antibodies use is as follows: anti-human Vβ5.1 (LC4, Invitrogen), anti-TCRβ (H57–597, BioLegend), anti-CD44 (IM7, BioLegend), anti-Vβ11 (KT11, BioLegend), anti-CD45.2 (104, BioLegend), anti-CD45.1 (A20, BioLegend), anti-SiglecF (E50–2440, BD Bioscience), anti-CD45 (30-F11, BioLegend), anti-CD11c (N418, BioLegend), anti-CD11b (M1/70, BioLegend), anti-Ly6G (1A8, BioLegend), anti-CD19 (6D5, BioLegend), anti-NK1.1 (PK136, BioLegend), anti-CD3 (17A2, BioLegend), Zombie Red (BioLegend), anti-mouse CD185/CXCR5 (L138D7, Biolegend), anti-mouse CD279/PD-1 (29F.1A12, Biolegend), and anti-Ki67 (SolA15, Invitrogen).
Morgun E., Zhu J., Almunif S., Bobbala S., Aguilar M.S., Wang J., Conner K., Cui Y., Cao L., Seshadri C., Scott E.A, & Wang C.R. (2023). Vaccination with mycobacterial lipid loaded nanoparticle leads to lipid antigen persistence and memory differentiation of antigen-specific T cells. bioRxiv.
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6

Comprehensive Flow Cytometry and ChIP Analysis

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For flow cytometry analysis, the following antibodies were used at a 1:100 dilution: anti-CD3 (17A2; Biolegend), anti-CD4 (GK1.5; Biolegend), anti-CD25 (PC61; Biolegend), anti-CD44 (IM7; Biolegend), anti-CD62L (MEL-14; Biolegend), anti-CD45.1 (A20; Biolegend), anti-CD45.2 (104; Biolegend), anti-FasL (MFL3; Biolegend), anti-IL-17A (TC11-18H10.1; Biolegend), anti-IFN-γ (XMG1.2; Biolegend), anti-JunB (C-11; Santa Cruz Biotechnology, Santa Cruz, CA, USA), anti-GATA3 (16E10A23; Biolegend), anti-ROR-γt (Q31-378; BD), anti-T-bet (4B10; Biolegend), and anti-rabbit IgG (Poly4064; Biolegend). For ChIP analyses, anti-JunB (1 μg per ChIP, 210; Santa Cruz), anti-BATF (1 μg per ChIP, WW8; Santa Cruz), anti-IRF4 (1 μg per ChIP, M-17; Santa Cruz) and rabbit IgG (1 μg per ChIP, I5006; Sigma) were used.
Hsieh T., Sasaki D., Taira N., Chien H., Sarkar S., Seto Y., Miyagi M, & Ishikawa H. (2022). JunB Is Critical for Survival of T Helper Cells. Frontiers in Immunology, 13, 901030.
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7

Monoclonal Antibodies for Hematopoietic Cell Analysis

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The following monoclonal antibodies (Abs) were used for flow cytometry and cell sorting: anti-c-Kit Microbeads (Miltenyi Biotec), anti-c-Kit (2B8, BD Biosciences / Biolegend), anti-Sca-1 (E13–161.7, BD Biosciences / Biolegend), anti-CD45.1 (A20, Biolegend), anti-CD45.2 (104, BD Biosciences), anti-CD4 (RM4–5, BD Biosciences), anti-B220 (RA3–6B2, BD Biosciences), anti-TER-119 (TER-119, Biolegend), anti-Gr-1 (RB6–8C5, BD Biosciences/Biolegend), anti-Mac-1 (M1/70, BD Biosciences), anti-CD71 (C2, BD Biosciences), anti-CD43 (S7, BD Biosciences), anti-CD16/32 (2.4G2, BD Biosciences), anti-Ki67 (B56, BD Biosciences), anti-CD34 (RAM34, eBioscience), anti-CD135 (A2F10, Biolegend), The primary Ab used for immunocytochemistry (ICC) was goat anti-Tfe3 (p-16; Santa Cruz Biotechnology). Anti-phospho mTor (Ser2448), anti-phospho Akt (Thr308), and anti-beta actin from Cell Signaling were used for western blotting.
Baba M., Toyama H., Sun L., Takubo K., Suh H.C., Hasumi H., Nakamura-Ishizu A., Hasumi Y., Klarmann K.D., Nakagata N., Schmidt L.S., Linehan W.M., Suda T, & Keller J.R. (2016). Loss of Folliculin disrupts hematopoietic stem cell quiescence and homeostasis resulting in bone marrow failure. Stem cells (Dayton, Ohio), 34(4), 1068-1082.
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8

Comprehensive Immune Cell Profiling

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For cell surface staining, cells were pre-incubated with 2.4G2 Fcγ RII/RIII blocking mAb for 15 min and then stained with the appropriate combinations of mAbs diluted in HBSS + 2% FBS for 30 min at 4 °C. Cells were analyzed on a BD FACS CantoII, or Cytek Aurora Spectral Cytometer, and data were processed with FlowJo software (TreeStar). The complete list of antibodies use is as follows: anti-human Vβ5.1 (LC4, Invitrogen), anti-TCRβ (H57-597, BioLegend), anti-CD44 (IM7, BioLegend), anti-Vβ11 (KT11, BioLegend), anti-CD45.2 (104, BioLegend), anti-CD45.1 (A20, BioLegend), anti-SiglecF (E50-2440, BD Bioscience), anti-CD45 (30-F11, BioLegend), anti-CD11c (N418, BioLegend), anti-CD11b (M1/70, BioLegend), anti-Ly6G (1A8, BioLegend), anti-CD19 (6D5, BioLegend), anti-NK1.1 (PK136, BioLegend), anti-CD3 (17A2, BioLegend), Zombie Red (BioLegend), anti-mouse CD185/CXCR5 (L138D7, Biolegend), anti-mouse CD279/PD-1 (29 F.1A12, Biolegend), and anti-Ki67 (SolA15, Invitrogen).
Morgun E., Zhu J., Almunif S., Bobbala S., Aguilar M.S., Wang J., Conner K., Cui Y., Cao L., Seshadri C., Scott E.A, & Wang C.R. (2023). Vaccination with mycobacterial lipid loaded nanoparticle leads to lipid antigen persistence and memory differentiation of antigen-specific T cells. eLife, 12, RP87431.
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9

Detailed Antibody Staining for Hematopoietic Cells

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The following antibodies (all from eBioscience, unless otherwise stated) were used: BM cell populations: anti-CD11b (M1/70) and anti-Gr1 (RB6-8C5). Assessment reciprocal BM chimeric mice: anti-CD45.1 (A20, BioLegend) and anti-CD45.2 (104). Hematopoietic stem and progenitor cells (HSPCs): anti-CD3ɛ (145-2C11), anti-CD4 (GK1.5, BioLegend), anti-CD8α (53-6.7, BioLegend), anti-B220 (RA3-6B2), anti-CD11b (M1/70), anti-Gr1 (RB6-8C5), anti-Ter119 (TER-119, BioLegend), anti-IL7Rα (A7R34), anti-NK1.1 (PK136, BioLegend), anti-cKit (2B8), anti–Sca-1 (D7, BioLegend), anti-CD34 (RAM34), anti-FcγR (93), anti-CD48 (HM48-1), and anti-CD150 (TC15-12F12.2, BioLegend). Classical and plasmacytoid dendritic cells: anti-CD3e (145-2C11), anti-CD19 (MB19-1), anti-NK1.1 (PK136; Becton Dickinson), anti-CD45RA (14.8; BD Biosciences), anti-CD11c (N418), and anti-MHCII (M5/114.15.2). BM non-hematopoietic BM stromal cells: anti-Sca1 (D7, BioLegend), anti-Ter119 (TER-119, BioLegend), anti-CD45 (30-F11, BioLegend), anti-CD31 (390), anti-CD105 (MJ7/18), and anti-CD140b (APB5). Interleukin-6 (IL-6) receptor α and β subunits: anti-CD126 (D7715A7, BioLegend) and anti-CD130 (KGP130).
Gerosa R.C., Boettcher S., Kovtonyuk L.V., Hausmann A., Hardt W.D., Hidalgo J., Nombela-Arrieta C, & Manz M.G. (2021). CXCL12-abundant reticular cells are the major source of IL-6 upon LPS stimulation and thereby regulate hematopoiesis. Blood Advances, 5(23), 5002-5015.
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10

Spleen Immunophenotyping After Infection

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Spleens from infected mice were harvested 7 d after infection and processed as described previously 66 . Sections were stained with anti-CD3 (17A2), anti-CD45.R/B220 (RA3-6B2) and anti-CD45.1 (A20; BioLegend). Spleen compartments were visualized by setting the density threshold of areas rich in CD3 (T cell zone) and B220 (B cell zone).
Scherer S., Oberle S.G., Kanev K., Gerullis A.K., Wu M., de Almeida G.P., Puleston D.J., Baixauli F., Aly L., Greco A., Nizharadze T., Becker N.B., Hoesslin M.V., Donhauser L.V., Berner J., Chu T., McNamara H.A., Esencan Z., Roelli P., Wurmser C., Kleiter I., Vehreschild M.J.G.T., Mayer C.A., Knolle P., Klingenspor M., Fumagalli V., Iannacone M., Prlic M., Korn T., Pearce E.L., Höfer T., Schulz A.M, & Zehn D. (2023). Pyrimidine de novo synthesis inhibition selectively blocks effector but not memory T cell development. Nature immunology, 24(3).
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