Cd45 hi30

Manufactured by BioLegend

Sourced in United States

CD45 (HI30) is a cell surface glycoprotein tyrosine phosphatase expressed on all hematopoietic cells except mature erythrocytes. It plays a crucial role in the regulation of a variety of cellular processes.

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Lab products found in correlation

Cd16 3g8, by BioLegend (4 mentions) Cd14 m5e2, by BioLegend (4 mentions) Hla dr l243, by BioLegend (4 mentions) Cd56 hcd56, by BioLegend (3 mentions) Cd4 okt4, by BioLegend (3 mentions) Mertk 2b10c42, by BioLegend (2 mentions) Ccr2 k036c2, by BioLegend (2 mentions) Lyve1 aly7, by Thermo Fisher Scientific (2 mentions) Bv421 sa, by BD (2 mentions) Cd64 10.1, by BioLegend (2 mentions) Ccr2 475301, by R&D Systems (2 mentions) Cd20 2h7, by BioLegend (2 mentions) Cd66b g10f5, by BioLegend (2 mentions) Cd11c 3, by Thermo Fisher Scientific (2 mentions) Cd8b 2st8.5h7, by Beckman Coulter (2 mentions) Cd123 6h6, by BioLegend (2 mentions) Attune nxt flow cytometer, by Thermo Fisher Scientific (2 mentions) Live dead fixable aqua dead cell stain kit, by Thermo Fisher Scientific (1 mentions) Cd80 2d10, by BioLegend (1 mentions) Cd3 sp34 2 and sk7, by BD (1 mentions)

Close spelling variants of this product

CD45 (clone HI30, (7 citations) CD45-BV421 (clone HI30, (4 citations) Anti-CD45 (clone HI30) (3 citations) Human CD45 (clone HI30, (3 citations) CD45-BV510 (clone HI30, (3 citations) PE-conjugated human CD45 antibody (clone HI30, (2 citations) PacBlue anti-CD45 (clone HI30); (2 citations) Anti-CD45-FITC mAbs (clone HI30; (2 citations) CD45-Pacific Blue (HI30) (2 citations) APC-CD45 (clone HI30, (2 citations)

8 protocols using cd45 hi30

Multiparameter Flow Cytometry Panel

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Cell suspensions were incubated with LIVE/DEAD Fixable Aqua Dead Cell Stain Kit (Life Technologies). Fc receptors were blocked using FcR block (Miltenyi) followed by staining with antibodies against surface molecules CD1c (AD5‐8E7; Miltenyi), CD3 (SP34‐2 and SK7; BD), CD11c (B‐Ly6; BD), CD14 (M5E2; BD), CD16 (3G8; Biolegend), CD19 (HIB19; Biolegend), CD20 (L27; BD), CD45 (HI30; Biolegend), CD56 (HCD56; Biolegend), CD66abce (TET2; Miltenyi), CD80 (2D10; Biolegend), CD123 (7G3; BD), CD141 (AD5‐14H12; Miltenyi), CCR7 (G043H7; Biolegend), and HLA‐DR (G46‐6; BD) for 15 min at 4°C in PBS with 2% FCS and fixed with 1% paraformaldehyde. Cells were analyzed using an LSRII flow cytometer (BD) and data were analyzed using FlowJo X software (Tree Star).

Lepzien R., Rankin G., Pourazar J., Muala A., Eklund A., Grunewald J., Blomberg A, & Smed‐Sörensen A. (2019). Mapping mononuclear phagocytes in blood, lungs, and lymph nodes of sarcoidosis patients. Journal of Leukocyte Biology, 105(4), 797-807.

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Multiparameter Immune Cell Profiling

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CD45 (HI30); MerTK (2B10C42); CD14 (M5E2); CD64 (10.1); CCR2 (K036C2); HLA-DR (L243).
All antibodies listed above were purchased from Biolegend with the exception of CCR2 (475301) (R&D Systems), Lyve1 (ALY7) (eBiosciences) and B220 (RA3–6B2); BV421-SA (BD Biosciences). Antibody clone is provided in parentheses. For more details see the Life Sciences Reporting Summary

Dick S.A., Macklin J.A., Nejat S., Clemente-Casares X., Momen A., Kantores C., Hosseinzadeh S., Barbu I., Chen J., Althagafi M.G., Besla R., Wong A., Aronoff L., Zaman R., Lavine K.J., Razani B., Ginhoux F., Husain M., Cybulsky M.I., Robbins C.S, & Epelman S. (2018). Self-renewing resident cardiac macrophages limit adverse remodeling following myocardial infarction. Nature immunology, 20(1), 29-39.

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Isolation and Purification of T-Cell Subsets

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Patients’ blood was collected in heparin tubes (40–50 ml in total), and peripheral blood mononuclear cells (PBMCs) were isolated by density gradient centrifugation using Ficoll density gradient medium (GE Healthcare Bio-Sciences AB, Uppsala, Sweden). CD4+ cells and CD8+ cells were isolated from the PBMCs via positive selection using CD4 or CD8 MicroBeads on an autoMACS® Pro Separator (Miltenyi Biotec Norden AB, Lund, Sweden). Flow cytometry was used to determine the purity of some of the sorted T-cell samples, and over 90% of CD45+ cells expressed CD4 (n = 5) or CD8 (n = 5). The following antibodies were used: CD45 (HI30; BioLegend, San Diego, CA, USA), CD4 (OKT4; BioLegend), and CD8 (SK1; BD Biosciences, Stockholm, Sweden).

Houtman M., Ekholm L., Hesselberg E., Chemin K., Malmström V., Reed A.M., Lundberg I.E, & Padyukov L. (2018). T-cell transcriptomics from peripheral blood highlights differences between polymyositis and dermatomyositis patients. Arthritis Research & Therapy, 20, 188.

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Multiparameter Immune Cell Profiling

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Characterization of Dermal APCs upon Plasmodium Stimulation

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Lysed single cell skin suspensions were counted and plated out in 48 well plates at 2*10⁵ cells per well. Cells were stimulated with mCherry expressing Pb or Wild-type Pf SPZ for 1 hour (Pb uptake experiments) or 24 hours (Pb and Pf stimulated dermal APC phenotyping). Cells were stained with 7AAD live/dead dye (uptake) or with Aqua fixable live/dead dye (Thermo Fischer Scientific; phenotyping), CD45 (HI30; Biolegend), HLA-DR (L243; eBioscience), CD11c (Bu15, Biolegend), PD-L1 (MIH1; BD Biosiences), CD80 (L307.4; BD Biosciences) and analyzed by Flow Cytometry using an LSRFortessa (BD Biosciences). Data was analyzed in FlowJo version 9.9.6 (FlowJo LLC). Gates were set using ‘fluorescence minus one’ (FMO) stained control samples.

Winkel B.M., Pelgrom L.R., van Schuijlenburg R., Baalbergen E., Ganesh M.S., Gerritsma H., de Korne C.M., Duszenko N., Langenberg M.C., Chevalley-Maurel S.C., Smits H.H., de Jong E.C., Everts B., Franke-Fayard B, & Roestenberg M. (2020). Plasmodium sporozoites induce regulatory macrophages. PLoS Pathogens, 16(9), e1008799.

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Multiparametric Phenotyping of Decidual Leukocytes

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1-2 X 10⁶ fresh decidual leukocytes were washed with PBS and stained using the following cocktail of antibodies: CD45 (HI30, Biolegend), CD66b (G10F5, BioLegend), CD20 (2H7, BioLegend), CD4 (OKT4, BioLegend), CD8b (2ST8.5H7, Beckman Coulter), CD14 (M5E2, BioLegend), HLA-DR (L243, BioLegend), CD56 (HCD56, Biolegend), CD16 (3G8, BioLegend), CD11c (3.9, ThermoFisher Scientific), CD123 (6H6, BioLegend), for 20 min in dark at 4C. Samples were washed twice in FACS buffer and resuspended in 400 uL. All samples were acquired with the Attune NxT Flow Cytometer (ThermoFisher Scientific, Waltham MA), immediately after addition of SYTOX Red Dead Cell Stain (1:1000). Data were analyzed using FlowJO (Ashland OR).

Sureshchandra S., Zulu M.Z., Doratt B.M., Jankeel A., Tifrea D., Edwards R., Rincon M., Marshall N.E, & Messaoudi I. (2022). Single-cell RNA sequencing reveals immunological rewiring at the maternal-fetal interface following asymptomatic/mild SARS-CoV-2 infection. Cell Reports, 39(11), 110938.

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Multiparameter Analysis of PBMC and Blister NK Cells

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Multiparameter analysis of PBMC and blister NK cell phenotype was performed on an ARIA II (BD Biosciences). PBMCs and blister cells were stained with different combinations of antibodies specific to human CD3(UCHT1;Biolegend), CD8(HIT8a;Biolegend), CD16(3G8;Biolegend), CD45(HI30;Biolegend), CD56(MEM-188;Biolegend), CD62L(DREG-56;Biolegend), CD69(FN50;Biolegend), CD107a(H4A3;Biolegend), NKG2D(1D11;Biolegend), CXCR6(K041E5;Biolegend), T-bet(04–46; BD), and EOMES(WD1928;ThermoFisher). All surface staining was performed for 30 min on ice after prior incubation of cells with Human Fc block and Murine Fc block (BD) for 10 minutes on ice. Following surface staining, cells were fixed and permeabilized using Foxp3 Transcription Factor Fixation/Permeabilization Concentrate and Diluent Kit (eBiosciences, Thermo Fisher), as per the manufacturers protocol, and stained for the expression of the transcription factors T-bet and Eomes. Fluorescence minus one control stains were performed using PBMC to verify the staining specificity and as a guide for setting markers to delineate positive and negative populations. Gating was set on the live lymphocyte population using forward- and side-scatter profiles to include lymphocytic blasts, followed by single cell gating using forward- and side-scatter heights and widths, before identification of hematopoietic cells using human CD45 staining.

Nikzad R., Angelo L.S., Aviles-Padilla K., Le D.T., Singh V.K., Bimler L., Vukmanovic-Stejic M., Vendrame E., Ranganath T., Simpson L., Haigwood N.L., Blish C.A., Akbar A.N, & Paust S. (2019). Human Natural Killer Cells Mediate Adaptive Immunity to Viral Antigens. Science immunology, 4(35), eaat8116.

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Comprehensive Immune Profiling of Decidual Leukocytes

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1–2 × 10⁶ fresh decidual leukocytes were washed with PBS and stained using the following cocktail of antibodies: CD45 (HI30, Biolegend), CD66b (G10F5, BioLegend), CD20 (2H7, BioLegend), CD4 (OKT4, BioLegend), CD8b (2ST8.5H7, Beckman Coulter), CD14 (M5E2, BioLegend), HLA-DR (L243, BioLegend), CD56 (HCD56, Biolegend), CD16 (3G8, BioLegend), CD11c (3.9, ThermoFisher Scientific), CD123 (6H6, BioLegend), for 20 minutes in dark at 4C. Samples were washed twice in FACS buffer and resuspended in 400 uL. All samples were acquired with the Attune NxT Flow Cytometer (ThermoFisher Scientific, Waltham MA), immediately after addition of SYTOX Red Dead Cell Stain (1:1000). Data were analyzed using FlowJO (Ashland OR).

Sureshchandra S., Zulu M.Z., Doratt B., Jankeel A., Tifrea D., Edwards R., Rincon M., Marshall N.E, & Messaoudi I. (2021). Deep immune profiling of the maternal-fetal interface with mild SARS-CoV-2 infection. bioRxiv.

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