Anti cd45.1 clone a20

Manufactured by BioLegend

Anti-CD45.1 (clone A20) is a mouse monoclonal antibody that recognizes the CD45.1 allotypic determinant expressed on the leukocyte common antigen. It is useful for identification and enumeration of CD45.1+ cells.

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Lab products found in correlation

Anti cd45.2 clone 104, by BioLegend (4 mentions) Anti cd44 clone im7, by BioLegend (4 mentions) Anti cd25 clone pc61, by BioLegend (3 mentions) Anti cd4 clone rm4 5, by BioLegend (3 mentions) Anti cd127 clone a7r34, by BioLegend (2 mentions) Anti foxp3 clone fjk 16s, by Thermo Fisher Scientific (2 mentions) Anti cd3 clone 145 2c11, by BioLegend (2 mentions) 4 6 diamidino 2 phenylindole (dapi), by Thermo Fisher Scientific (2 mentions) Anti b220 clone ra3 6b2, by BD (1 mentions) Anti cd45rb clone 16a, by BD (1 mentions) Anti cd8a clone 53 6.7, by BioLegend (1 mentions) Live dead fixable violet, by Thermo Fisher Scientific (1 mentions) Anti cd8α clone 53 6, by BioLegend (1 mentions) Zombie green fixable viability kit, by BioLegend (1 mentions) Anti cd5 clone 53 7, by BioLegend (1 mentions) Anti cd24 clone m1 69, by BioLegend (1 mentions) Anti cd8 clone mem 31, by Exbio (1 mentions) Anti klrg 1 clone 2f1, by BioLegend (1 mentions) Anti ly6c clone hk1, by BioLegend (1 mentions) Hoechst 33258, by Thermo Fisher Scientific (1 mentions)

6 protocols using anti cd45.1 clone a20

Flow Cytometry Analysis of Mouse Immune Cells

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The following anti-mouse antibodies were used for flow cytometric analysis: anti-CD4 (clone RM4-5, Biolegend); anti-CD8a (clone 53-6.7, Biolegend); anti-CD45.1 (clone A20, Biolegend); anti-CD45.2 (clone 104, Biolegend); anti-CD25 (clone PC61, Biolegend); anti-CD45RB (clone 16A, BD Biosciences); anti-B220 (clone Ra3-6B2, BD Biosciences); anti-CD11c (clone L3, BD Biosciences); anti-Thy1.1 (clone OX-7, Biolegend); anti-Thy1.2 (clone 53-2.1, Biolegend); anti-CCR7 (clone 4B12, Biolegend). Antibodies for total eIF2α and p-Ser51 eIF2α were from cell Signaling (clones D7D3 and D9G8 rabbit XP mAbs). OVA 257–264 and OVA 323–339 peptides were purchased from American Peptide Company. Live/Dead Fixable Violet was purchased from Invitrogen.

Van de Velde L.A., Guo X.Z., Barbaric L., Smith A.M., Oguin TH I.I.I., Thomas P.G, & Murray P.J. (2016). Stress kinase GCN2 controls the proliferative fitness and trafficking of cytotoxic T cells independent of environmental amino acid sensing. Cell reports, 17(9), 2247-2258.

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Flow Cytometry Antibody Panel for Immune Cell Profiling

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Antibodies to following antigens were used for flow cytometry: anti-CD4 (clone RM4-5, Biolegend #100536), anti-CD5 (clone 53-7-3, Biolegend #100627, #100625), anti-CD8α (clone 53-6.7, Biolegend #100738), anti-CD24 (clone M1/69, Biolegend #101806), anti-CD25 (clone PC61, Biolegend #102016 and #102036), anti-CD44 (clone IM7, Biolegend #103049), anti-CD45.1 (clone A20, Biolegend #110723), anti-CD45.2 (clone 104, Biolegend #109808), anti-CD49d (clone R1-2, Biolegend #103618), anti-CD127 (clone A7R34, Biolegend #135012), anti-TCRβ (clone H57-597, Biolegend #109218), anti-KLRG1 (clone 2F1, Biolegend #138421), anti-Ly6C (clone HK1.4, Biolegend #128006), biotin-conjugated anti-BST2 (clone 927, Biolegend #127006), anti-biotin (clone 1D4-C5, Biolegend #409004). Viability was stained with LIVE/DEAD™ Fixable Near-IR Dead Cell Stain Kit (Invitrogen #L34975), Zombie Green™ Fixable Viability Kit (Biolegend, #423111) or Hoechst 33258 (Invitrogen, #H3569). For the analysis of Jurkat cell lines, anti-CD8 (clone MEM-31, Exbio #1P-207-T025), anti-CD69 (clone FN50, Exbio #T7-552-T100) were used. Antibodies were conjugated with various fluorophores and used according to the manufacturer’s instructions.

Paprckova D., Niederlova V., Moudra A., Drobek A., Pribikova M., Janusova S., Schober K., Neuwirth A., Michalik J., Huranova M., Horkova V., Cesnekova M., Simova M., Prochazka J., Balounova J., Busch D.H., Sedlacek R., Schwarzer M, & Stepanek O. (2022). Self-reactivity of CD8 T-cell clones determines their differentiation status rather than their responsiveness in infections. Frontiers in Immunology, 13, 1009198.

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Comprehensive Immune Cell Profiling by Flow Cytometry

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Flow cytometry was performed as previously described Fang et al. (2010 (link)). The following antibodies were used: anti-CD3e (clone 145-2C11, Biolegend), anti-CD11b (clone M1/70, Biolegend), anti-CD19 (clone 1D3, BD Biosciences), anti-CD27 (clone LG.3A10, Biolegend), anti-CD29 (clone HMβ1-1, Biolegend San Diego, CA), anti-CD44 (clone IM7, Biolegend), anti-CD45.1 (clone A20, Biolegend), anti-CD45.2 (clone 104, Biolegend), anti-CD49a (clone HMα1, Biolegend), anti-CD49b (clone DX5, Biolegend), anti-CD105 (clone MJ7/18, Biolegend), anti-CD106 (clone 429, Biolegend), anti-BrDU (clone PRB-1, eBioscience), anti-CXCR3 (clone CXCR3-173, Biolegend), anti-Eomes (clone Dan11mag, eBioscience), anti-KLRG1 (clone 2F1/KLRG1, Biolegend), anti-Ly-6A/E (Sca-1) (clone E13-161.7, Biolegend), anti-Ly49A (clone A1/Ly49A, Biolegend), anti-Ly49C/I (clone 5E6, Biolegend), anti-Ly49D (clone 4E5, Biolegend), anti-Ly49G2 (clone LGL-1, eBioscience), anti-Ly49H (clone 3D10, Biolegend), anti-NK1.1 (clone PK136, Biolegend), NKG2A/C/E (clone 20d5, Biolegend), anti-Tbet (clone 4B10, Biolegend), and anti-TRAIL (clone N2B2, Biolegend). At least 500 000 cells were analyzed by flow cytometry at the Fox Chase Cell Sorting Facility using an LSR II system (BD Biosciences).

Nair S., Fang M, & Sigal L.J. (2014). The natural killer cell dysfunction of aged mice is due to the bone marrow stroma and is not restored by IL-15/IL-15Rα treatment. Aging Cell, 14(2), 180-190.

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Multiparametric Flow Cytometry Analysis

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Blood or brain single-cell suspensions were washed in PBS supplemented with 1% FBS, 2 mM EDTA and 0.01% sodium azide, and incubated for 10 min at 4 • C with culture supernatant from the hybridoma cell line 2.4G2 to block Fc receptors. The cells were surface stained with fluorochrome-conjugated monoclonal antibody for 15 min at 4 • C. LIVE/DEAD fixable blue dye (Life Technologies) was used to exclude dead cells. The following antibodies were used: anti-CD45.1 (clone A20, BioLegend), anti-CD11b (clone M1/70, BD), anti-CD19 (clone 6D5, BioLegend), anti-CD3 (clone 145-2C11, BioLegend), anti-CD4 (clone RM4-5, BD), anti-CD8 (clone 53-6.7, BD or Biolegend), anti-FOXP3 (clone FJK-16s, eBioscience), anti-CD44 (clone IM7, BioLegend), anti-CD62L (clone MEL-14, BD), anti-CD127 (clone A7R34, BioLegend) and anti-KLRG1 (clone 2F1, eBioscience). OVA-specific CD8 + T cells were detected using H-2K b /SIINFEKL pentamers (ProImmune). Intracellular FoxP3 staining was performed using BioLegend FOXP3 Fix/Perm Buffer Set. Samples were acquired on LSRII Flow Cytometer (BD), and the data were analyzed using FlowJo software (TreeStar Inc.).

Grasso C., Field C.S., Tang C.W., Ferguson P.M., J Compton B., Anderson R.J., Painter G.F., Weinkove R., F Hermans I, & Berridge M.V. (2020). Vaccines adjuvanted with an NKT cell agonist induce effective T-cell responses in models of CNS lymphoma. Immunotherapy, 12(6).

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Fluorescent Nanoparticle Synthesis and Characterization

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ReCl₃·6H₂O (Er, Yb, Y > 99%), oleic acid (OA), oleylamine (OM), 1-octadecene (ODE), yttrium trifluoroacetate ((CF₃COO)₃Y), sodium trifluoroacetate (CF₃COONa), sodium hydroxide (NaOH), ammonium fluoride (NH₄F), N-hydroxy-succinimide (NHS), 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide (EDC), fluorescein isothiocyanate (FITC), m ethanol, ethanol, acetone, cyclohexane, N,N-dimethylformamide (DMF), dimethyl sulfoxide (DMSO), and polyacrylic acid (PAA) were purchased from Sigma-Aldrich. All chemicals were used without further purification. DAPI and anti-CD86-FITC (clone GL1) were purchased from Thermo Fisher. Anti-CD69-PE (clone H1.2F3), anti-CD62L-APC (clone MEL-14), and anti-CD3 (clone 145-2C11) were purchased from BioLegend. Alpha-Amino-omega-carboxy poly(ethylene glycol) hydrochloride (H₂N-PEG-COOH*HCL, MW 3.000 Dalton) was purchased from Iris Biotech. Cell Titer 96 AQueous MTS Reagent Powder was purchased from Promega. Anti-CD45.1 (clone A20), CD8α + (clone 53–6,7), and anti-CD43 (clone 1B11) were purchased from BioLegend.

Yu Z., Vepris O., Eich C., Feng Y., Que I., Camps M.G., Zhang H., Ossendorp F.A, & Cruz L.J. (2022). Upconversion nanoparticle platform for efficient dendritic cell antigen delivery and simultaneous tracking. Mikrochimica Acta, 189(10), 368.

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Profiling Murine Splenic Immune Cells

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Mouse spleen cells were washed with PBS with 1% BSA (staining buffer), blocked with mouse IgG in staining buffer, then stained with conjugated antibodies. Antibodies used were anti-CD4 (clone RM4-5, Biolegend, San Diego, CA), anti-CD25 (clone PC61, Biolegend), anti-PD-1 (clone 29F.1A12, Biolegend), anti-PD-L1 (clone 10F.9G2 Biolegend), anti-Nrp-1 (Cat FAB5994N, R&D Systems, Minneapolis, MN), anti-FoxP3 (clone FJK-16s, eBiosciences), anti-CD44 (clone IM7, Biolegend), anti-CD62L (clone MEL-14, Biolegend), anti-CD45.1 (clone A20, Biolegend), and anti-CD45.2 (clone 104, Biolegend).
Stained cells were analyzed in the Oklahoma Medical Research Facility (OMRF) Flow Cytometry Core Facility on a BD LSRII (BD Biosciences) data was analyzed using FlowJo Software (Tree Star, Inc., Ashland, OR).

Muhammad F., Avalos P.N., Mursalin M.H., Ma J.X., Callegan M.C, & Lee D.J. (2020). Kallistatin Attenuates Experimental Autoimmune Uveitis by Inhibiting Activation of T Cells. Frontiers in Immunology, 11, 975.

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