Cd45.1 clone a20

Manufactured by BD

Sourced in United States

CD45.1 (Clone A20) is a lab equipment product that functions as a cell surface marker. It is used to identify and distinguish specific cell types in various biological research applications.

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Lab products found in correlation

Cd45.2 clone 104, by BioLegend (4 mentions) Cd45 clone 104, by BioLegend (3 mentions) Cd45.2 clone 104, by BD (3 mentions) Facsaria 3, by BD (2 mentions) Sca 1, by BioLegend (2 mentions) Lsrii flow cytometer, by BD (2 mentions) Cd8 clone 53 6, by BD (2 mentions) Ly6c clone al 21, by BD (2 mentions) Cd150 clone mshad150, by Thermo Fisher Scientific (2 mentions) Cd48 clone hm48 1, by BioLegend (2 mentions) Lineage cocktail, by BioLegend (1 mentions) Diva software, by BD (1 mentions) Cd44 clone im7, by BD (1 mentions) Cd62l mel 14, by BD (1 mentions) Cd45.2 clone 104, by Thermo Fisher Scientific (1 mentions) Rosettesep human t cell enrichment cocktail, by STEMCELL (1 mentions) Cd90 clone 53 2.1, by BD (1 mentions) Cd45rb clone c363.16a, by Thermo Fisher Scientific (1 mentions) Ccr7 clone 150503, by BD (1 mentions) Brefeldin a golgiplug, by BD (1 mentions)

Close spelling variants of this product

CD45.1 (A20; (13 citations) Anti-CD45.1 (clone A20) (3 citations) Anti-CD45.1-PE (clone A20; (2 citations)

10 protocols using cd45.1 clone a20

Multiparametric Flow Cytometry for Hematopoietic Stem Cells

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All samples were analyzed by flow cytometry using FACSAria III or LSR (BD biosciences, San Jose, CA, USA). CD45 (Clone 104) (Biolegend, San Diego, CA, USA), CD45.1 (Clone A20) (BD Biosciences), CD45.2 (Clone 104) (Biolegend), Lineage cocktail (include CD3, B220, Gr1, CD11b, Ter119; Biolegend), cKit (Clone 2B8, Biolegend), Sca1 (Clone D7, Biolegend), EPCR (CD201) (Clone eBio1560, eBioscience, San Diego, CA, USA), CD150 (Clone mShad150, eBioscience), and CD48 (Clone HM48-1, Biolegend) antibodies were used.

Castiglione M., Zhang H., Kaushansky K, & Zhan H. (2021). Cell competition between wild-type and JAK2V617F mutant cells in a murine model of a myeloproliferative neoplasm. Experimental hematology, 100, 52-62.

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Comprehensive T Cell Phenotyping Protocol

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Human cells were stained with fluorescence-conjugated antibodies to CD3 (clone HIT3a), CD4 (clone RPA-T4), CD8 (clone RPA-T8), CD45RA (clone HI100), CD45RO (clone UCHL1), CD62L (clone DREG–56), CCR7 (clone 150503) (BD Biosciences), and CD7 (clone 124-1D1, eBioscience). T cell subsets were enriched by negative selection with a RosetteSep Human T Cell Enrichment Cocktail (StemCell Technologies) from peripheral blood and ovarian cancer tissues. Naïve T cells were obtained with CD45RO bead–negative selection (Miltenyi Biotec) and sorted and confirmed by FACS with 98% purity. Mouse single-cell suspensions were prepared from thymus, bone marrow, peripheral blood, lymph nodes, and spleen and labeled with fluorescence-conjugated antibodies to CD3 (clone 17A2), CD4 (clone RM4–5), CD8 (clone 53-6.7), CD19 (clone 1D3), CD44 (clone IM7), CD45.1 (clone A20), CD62L (MEL–14), CD90 (clone 53-2.1) (BD Biosciences), CD45.2 (clone 104), and CD45RB (clone C363.16A, eBioscience). Then, if necessary, the cells were sorted (BD FACSAria) or acquired on an LSR II flow cytometer (BD Biosciences), and data were analyzed with DIVA software (BD Biosciences).

Xia H., Wang W., Crespo J., Kryczek I., Li W., Wei S., Bian Z., Maj T., He M., Liu R.J., He Y., Rattan R., Munkarah A., Guan J.L, & Zou W. (2017). Suppression of FIP200 and autophagy by tumor-derived lactate promotes naïve T cell apoptosis and affects tumor immunity. Science immunology, 2(17), eaan4631.

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Multiparametric Flow Cytometry of Immune Cells

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Cell cultures were performed in complete culture medium consisting of RPMI-1640 supplemented with 10% heat-inactivated FCS, 2 mM L-glutamine (Sigma, St. Louis, MO), 50 μg/ml gentamicin (PAA, Linz, Austria), and 50 μM beta-mercaptoethanol (Sigma).
Phenotypical analyses were performed by flow cytometry with mAb against CD4 (clone RM4-5), CD45.1 (clone A20), MHC class II (anti-I-A/I-E^diverse, clone 2G9), CD11c (clone HL3), CD8α (clone Ly-2), CD45 (clone 30-F11), CD103 (clone M290), EpCAM/CD326 (clone G8.8), CD19 (clone 1D3), NK1.1 (clone PK136), IL-12p40 (clone C15.6) (all from BD-Pharmingen, San Diego, CA), CD127 (clone A7R34, Biolegend, San Diego, CA), CD70 (clone FR70, Biolegend), and Langerin/CD207 mAb (clone 929F3; Dendritics, Lyon, France). When possible, viable cells were determined by exclusion of 7-AAD-positive dead cells (BD-Pharmingen). IL-12p40 and IL-10 stainings were performed on total lymph node cell suspension (10⁶ cells/ml) incubated for 3 h in 1 μg/ml Brefeldin A (Golgiplug, BD-Pharmingen). To stain for Langerin or intracellular cytokines, permeabilization was performed with Cytofix/perm kit (BD-Pharmingen).

Flacher V., Tripp C.H., Mairhofer D.G., Steinman R.M., Stoitzner P., Idoyaga J, & Romani N. (2014). Murine Langerin+ dermal dendritic cells prime CD8+ T cells while Langerhans cells induce cross-tolerance. EMBO Molecular Medicine, 6(9), 1191-1204.

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Isolation and Characterization of Lung Immune Cells

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BALF was recovered from mice at the specified timepoint in 1 ml sterile PBS containing 5 mM EDTA as previously described [90 (link)]. Cells were surface stained 30 min in cold FACS buffer (PBS with 2.5% FBS, 5 mM EDTA) with Siglec-F (clone E50-2440, Biolegend), CD11c (clone N418, Biolegend), Ly6G (clone 1A8, BD Biosciences), Ly6C (clone AL-21, BD Biosciences, Allschwil, Switzerland), CD11b (clone M1/70, Biolegend), CD45.1 (clone A20, BD Biosciences), CD45.2 (clone 104, BD Biosciences).
For intracellular staining of TNF (clone MP6-XT22, Biolegend), mice were injected i.p. with 50 μl of 5 mg/ml Brefeldin A in EtOH (diluted with 100 μl PBS) 3 hours prior to taking BALF. Lavage was performed with 1 ml PBS 5mM EDTA containing 5 μg/ml Brefeldin A, and was immediately placed on ice. After surface stain, cells were washed with FACS buffer and fixed, permeabilized and stained using the BD Biosciences Cytofix/Cytoperm Kit according to the manufacturer's instructions. Data were acquired on an LSRII (BD Biosciences) and analyzed with FlowJo software (TreeStar, Ashland, OR). An Aria III instrument (BD Biosciences) was used for cell sorting.

Ziltener P., Reinheckel T, & Oxenius A. (2016). Neutrophil and Alveolar Macrophage-Mediated Innate Immune Control of Legionella pneumophila Lung Infection via TNF and ROS. PLoS Pathogens, 12(4), e1005591.

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Multiparametric flow cytometry for hematopoietic stem cells

Cited 1 time

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All samples were analyzed by flow cytometry using a FACSAriaTM III (BD biosciences, San Jose, CA, USA). CD45 (Clone 104) (Biolegend, San Diego, CA, USA), CD45.1 (Clone A20) (BD Biosciences), CD45.2 (Clone 104) (Biolegend), EPCR (CD201) (Clone eBio1560, eBioscience, San Diego, CA, USA), CD150 (Clone mShad150, eBioscience), and CD48 (Clone HM48–1, Biolegend) antibodies were used to enumerate CD45⁺EPCR(CD201)⁺CD48⁻CD150⁺ (E-SLAM) cells.[22 (link),23 (link)] MKs were sorted on the basis of CD41 expression (Clone MWReg30, Biolegend) and cell size.[15 (link)] CD45 (Clone 104) (Biolegend) and CD31 (Clone 390, BD biosciences) antibodies were used to isolate CD45⁻CD31⁺ marrow ECs.[24 (link)]

Zhang Y., Lin C.H., Kaushansky K, & Zhan H. (2018). JAK2V617F Megakaryocytes Promote Hematopoietic Stem/Progenitor Cell Expansion in Mice through Thrombopoietin/MPL signaling. Stem cells (Dayton, Ohio), 36(11), 1676-1684.

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Multiparametric Immune Cell Profiling

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Lymphocytes were counted and subjected to viability staining (Fixable Aqua Dead Cell staining, Life Technologies, #L34957) and subsequently to receptor Fc blockade (BD #553142). Staining was performed using the following antibodies: CD4 (clone RM4-4, BD), CD45 (clone 30-F11 eBioscience), CD45.1 (clone A20, BD), CD45.2 (clone 104, BD) CD8a (clone 53-6.7, eBioscience or BD), CD8b (clone YTS156.7.7, BioLegend), CD11b (clone RM2817, Thermo Fisher), Ly6c (clone AL-21, BD), CD3ε (clone 145-2C11, BD), CD19 (clone 1D3, BD), CD90.2 (53-2.1, BD), CD127 (clone A7R34, BD), CX3CR1 (clone SA011F11, BioLegend), CXCR3 (clone CXCR3-173, Biolegend), CD103 (clone 2E7, eBioscience or Biolegend), CD69 (clone H1.2F3, BD), TCR-β (clone H57-597, BD, TCR-γδ (clone eBio-GL3, eBioscience). H2-K^b:SIINFEKL or H2-M3:fMIGWII tetramers were either produced in house or, for the former, obtained thorugh the NIH tetramer core. Samples were fixed (IC Fixation Buffer, eBioscience), washed, resuspended in FACS buffer and acquired with a LSRII flow cytometer (BD) either immediately or on the following day. For intracellular staining, the following antibodies were used: IFN-ϒ (clone XMG1.2, BD), TNF-α (clone MP6-XT22, BD) in Permeabilization buffer (eBioscience).

Becattini S., Littmann E.R., Seok R., Amoretti L., Fontana E., Wright R., Gjonbalaj M., Leiner I.M., Plitas G., Hohl T.M, & Pamer E.G. (2020). Enhancing mucosal immunity by transient microbiota depletion. Nature Communications, 11, 4475.

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Characterization of Hematopoietic Stem Cells

Cited 1 time

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All samples were analyzed by flow cytometry using a CytoFLEX flow cytometer (Beckman Coulter, Indianapolis, IN, USA). CD45 (Clone 104) (Biolegend, San Diego, CA, USA), CD45.1 (Clone A20) (BD Biosciences, San Jose, CA), CD45.2 (Clone 104) (Biolegend), Lineage (Lin) cocktail (include CD3, B220, Gr1, CD11b, Ter119; Biolegend), cKit (Clone 2B8, Biolegend), Sca1 (Clone D7, Biolegend), CD150 (Clone mShad150, eBioscience, San Diego, CA, USA), CD48 (Clone HM48-1, Biolegend), CD31 (Clone 390, BD Biosciences), and Ter119 (Clone Ter-119, Biolegend) antibodies were used. Intracellular CXCL12 expression and membranal SCF expression were analyzed using an anti-mouse CXCL12 antibody (Clone 79018, R&D Systems, Minneapolis, MN, USA) and an anti-mouse SCF antibody (Clone #40215, R&D Systems) as we previously described (Zhan et al., 2018 (link)).

Lee S, & Zhan H. (2024). Deciphering the differential impact of thrombopoietin/MPL signaling on hematopoietic stem/progenitor cell function in bone marrow and spleen. Stem Cell Reports, 19(2), 211-223.

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Multicolor Flow Cytometry Analysis

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Antibodies against B220 (clone RA3-6B2), CD3 (clone 145-2C11), CD4 (clone GK1.5), CD8 (clone 53-6.7), CD11b (clone M1/70), CD45.1 (clone A20), CD45.2 (clone 30-F11), H-2K d (clone SF1-1.1), H-2K b (clone 25-D1.16), and IA/IE (clone M5/114) labeled with various fluorescent dyes, their appropriate isotype controls, and 7aminoactinomycin D (7-AAD) were purchased from BD Biosciences (Franklin Lakes, NJ, USA). Cells were analyzed using a BD fluorescence-activated cell sorting (FACS) Canto II Flow Cytometer and BD FACS Diva software (BD Biosciences). Cells positive for 7-AAD staining were excluded from the analysis as nonviable cells.

Igarashi K., Hori T., Yamamoto M., Sohma H., Suzuki N., Tsutsumi H., Kawasaki Y, & Kokai Y. (2022). CCL8 deficiency in the host abrogates early mortality of acute graft-versus-host disease in mice with dysregulated IL-6 expression. Experimental hematology, 106.

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Bone Marrow Transplant Reconstitution Assay

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Young CD45.2 WT or mPGES-1 KO female mice were used as donors. Bone marrow was harvested under aseptic conditions from the femur, tibia, and iliac of donor mice. Erythrocytes were lysed and the remaining cells were used for bone marrow transplant. Recipient mice were lethally irradiated using 2 doses of 5 Gy on the same day. After the last irradiation, the mice received 5 million donor cells via tail vein. Mice received ampicillin (2 g/L in drinking water) for 6 weeks, and bone marrow reconstitution was assessed after 2 months using the expression of CD45.2 marker by flow cytometry. The following antibodies were used: CD45.1 clone A20 from BD biosciences and CD45.2 clone 104 from Biolegend. Samples were run on a BD LSR II flow cytometer and analyzed with Flowjo (Treestar) software.

Peña Silva R.A., Mitchell I.J., Kung D.K., Pewe L.L., Granja M.F., Harty J.T., Faraci F.M., Heistad D.D, & Hasan D.M. (2015). Paradoxical Increase in Mortality and Rupture of Intracranial Aneurysms in mPGES-1 Deficient Mice: Attenuation by Aspirin. Neurosurgery, 77(4), 613-620.

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LPS Induction of Immune Responses

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Escherichia coli O111 LPS was from Sigma-Aldrich (L2630, purified by phenol extraction; St. Louis, MO, USA). Neisseria meningitidis LPS, purified from a group B (L3,7,9) strain, was a generous gift from M. Apicella (University of Iowa Medical Center, Iowa City, IA, USA). [20] [21] [22] E. coli O14 was obtained from the Statens Serum Institut (Copenhagen, Denmark) and its LPS (Ra chemotype) 23 was prepared by the phenol-chloroform-light petroleum method. 24 The ability of each of the LPS preparations to induce TNF-a production was at least 20-fold less in TLR4 À/À macrophages than in TLR4 +/+ macrophages. E. coli O14 LPS-FITC was made as described. 25, 26 OVA-Alexa 594 was from Life Technologies (Carlsbad, CA, USA). Abs used for flow cytometry: anti-CD69 (Clone H1.2F3), anti-CD86 (Clone GLI), B220 (Clone RA3-6B2), CD45.1 (Clone A20) and CD45.2 (Clone 104) were from BD Biosciences (Franklin Lakes, NJ, USA).

Lu M, & Munford R. (2016). LPS stimulates IgM production in vivo without help from non-B cells. Innate immunity, 22(5).

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