Anti cd48

Manufactured by BioLegend

Anti-CD48 is an antibody product offered by BioLegend. It recognizes the CD48 antigen, which is a cell surface glycoprotein involved in various cellular processes. The core function of this product is to detect and bind to the CD48 antigen, enabling its use in research applications.

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Lab products found in correlation

Close spelling variants of this product

Anti-CD48 FITC (2 citations) Anti-CD48 (HM48-1), (2 citations) ), anti-CD48-Alexa Fluor 647 (HM48–1; 103416) (2 citations)

5 protocols using anti cd48

Phenotypic Profiling of Myeloid Lineages

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Whole blood was stained with directly-conjugated antibodies and incubated for 30 mins at 4 °C. Following red blood cell lysis (BD FACS Lysing solution), myeloid lineages were identified using anti-CD45, anti-CD14, anti-CD11b (BD Pharmingen), anti-CD16 (eBioscience), and anti-CD62L (Beckman Coulter)(online resource 1A, B). For analysis of SLAM family receptor expression, anti-CD150, anti-CD84, anti-CD229 (eBioscience), anti-CD48, anti-CD244, anti-CD352 and anti-CD319 (Biolegend) antibodies were used. Acquisition and analysis were performed using a BD LSRFortessa™ and FlowJo v10.1 for Windows, respectively (Treestar). Median fluorescence intensity (MFI) was normalised to a mean equivalent of fluorochrome (MEF) SPHERO™ Rainbow Calibration Particles and the isotype control, as per the manufacturer’s instructions (online resource 1C).

Mak A., Thornhill S.I., Lee H.Y., Lee B., Poidinger M., Connolly J.E, & Fairhurst A.M. (2017). Brief report: Decreased expression of CD244 (SLAMF4) on monocytes and platelets in patients with systemic lupus erythematosus. Clinical Rheumatology, 37(3), 811-816.

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Immunophenotyping of Hematopoietic Progenitors

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The dissociated cells were incubated with antibody madoptixture at 4 °C for 30 min, followed by incubation with 7-AAD antibody at room temperature for 5 min. Cells were sorted and analyzed by flow cytometers FACS Aria II (BD Biosciences) and MoFlo XDP (Beckman Coulter) in the purity model. The FACS data were analyzed with FlowJo software (v10, Tree star). Surface markers for E10.0 early AECs in AGM regions were CD41⁻CD43⁻CD45⁻CD31⁺CD44⁺Kit⁻. Surface markers for E10.0 HECs in AGM regions were CD41⁻CD43⁻CD45⁻CD31⁺CD44⁺Kit⁺CD201⁺. Surface markers of E11.0 AGM pre-HSCs were CD31⁺Kit⁺CD201⁺. Surface markers for E14.5 fetal liver LT-HSCs were CD45⁺CD201⁺CD150⁺CD48⁻. Cells were stained using the following antibodies (1:100 diluted): Anti-CD31 (BD, MEC13.3, Catalog# 562939), Anti-CD41 (BD, MWReg30, Catalog# 553848), Anti-CD43 (BD, S7, Catalog# 553270), Anti-CD44 (BioLegend, Catalog# 103044), Anti-CD45 (BD, 30-F11, Catalog# 553079), Anti-CD48 (BioLegend, Catalog# 103432), Anti-CD201 (eBioscience, eBio1560, Catalog# 17-2012-82), Anti-Kit (eBioscience, 2B8, Catalog# 14-1171-82), and Anti-CD150 (BioLegend, Cat# 115904). 7-aminoactinomycin D (7-AAD; eBioscience, Catalog# 00-6993-50) was used to exclude dead cells. The FACS Diva 8 “index sorting” function was activated and sorting was performed in the single-cell mode.

Li C.C., Zhang G., Du J., Liu D., Li Z., Ni Y., Zhou J., Li Y., Hou S., Zheng X., Lan Y., Liu B, & He A. (2022). Pre-configuring chromatin architecture with histone modifications guides hematopoietic stem cell formation in mouse embryos. Nature Communications, 13, 346.

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Multiparametric Flow Cytometry Analysis

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Human or mouse cells were harvested and Fc receptors of cells were blocked with anti-CD16/32 (BioLegend) or human IgG (Sigma-Aldrich) for 10 min followed by staining of cells for 15 min at room temperature. For intracellular staining, cells were fixed in 2% formaldehyde and then permeabilized using the Perm/Wash buffer kit (BD Biosciences) followed by antibody incubation. Monoclonal antibodies used for flow cytometry include the following: anti–c-Kit, anti-Sca1, anti-CD48, anti-CD150, anti-Flt3 ligand, anti-CD3, anti-TER119, anti–Gr-1, and anti-B220 (BioLegend) for mouse studies and anti-CD68 (BioLegend), anti-CD163 (Biolegend), and anti-206 (eBioscience) for human studies. Rabbit antitrimethylated H3K4 (Abcam) and antitrimethylated H3K27 (Millipore) were used to detect histone marks, followed by secondary stain with FITC-AffiniPure goat anti-rabbit IgG (Jackson ImmunoResearch). All populations were routinely back gated to verify purity and gating. Samples were analyzed on an LSR II (BD Biosciences). One million viable cells were analyzed. Data were analyzed using FlowJo software version 9.0 (Tree Star, Inc.) and compiled using Prism (GraphPad Software).

Gallagher K.A., Joshi A., Carson W.F., Schaller M., Allen R., Mukerjee S., Kittan N., Feldman E.L., Henke P.K., Hogaboam C., Burant C.F, & Kunkel S.L. (2014). Epigenetic Changes in Bone Marrow Progenitor Cells Influence the Inflammatory Phenotype and Alter Wound Healing in Type 2 Diabetes. Diabetes, 64(4), 1420-1430.

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Phenotypic Characterization of Lymphocytes

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Cells isolated from spleens and graft-draining axillary and brachial lymph nodes (dLN) were stained with anti-CD4 (BD Biosciences), anti-CD8 (Invitrogen) and anti-Thy1.1 (BD Biosciences). For phenotypic analysis cells were also surface-stained with anti-PD-1 (BioLegend), anti-2B4 (BD Biosciences or eBioSciences), anti-Thy1.1 (BD Biosciences), anti-LAG-3 (BioLegend), anti-CD127 (BioLegend), anti-KLRG-1 (eBioSciences), anti-CD44 (BioLegend or BD Biosciences), and anti-CD48 (BioLegend). Absolute numbers of lymphocytes from the spleen and draining lymph nodes were calculated using a Cellometer Auto T4 Cell Viability Counter (Nexcelom) according to the manufacturer’s instructions. Samples were analyzed on an LSRII flow cytometer (BD Biosciences). Data was analyzed using FlowJo 9 software (Treestar, San Carlos, CA) and Prism 6 software (GraphPad Software Inc.). For intracellular cytokine staining, lymphocytes were restimulated ex vivo with 1 μg/mL phorbol 12-myristate 13-acetate (PMA) (Sigma Life Sciences) and 1 μg/mL ionomycin (Sigma Life Sciences) where indicated, in the presence of 1 μg/mL Brefeldin A (BD Biosciences) for 4 hours. The Fix/Perm intracellular staining kit (BD Pharmingen) was used to detect IL-2 (BD Biosciences), TNF (BioLegend), and IFN-γ (BD Biosciences), according to manufacturer’s instructions.

Laurie S.J., Liu D., Wagener M.E., Stark P.E., Terhorst C, & Ford M.L. (2018). 2B4 Mediates Inhibition of CD8+ T Cell Responses via Attenuation of Glycolysis and Cell Division. Journal of immunology (Baltimore, Md. : 1950), 201(5), 1536-1548.

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Immunohistochemistry of Bone Tissue Sections

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Frozen tissue sections prepared from 4% paraformaldehyde-fixed and decalcified bones were thawed at room temperature and then rehydrated with PBS. The slides were stained with the following antibodies (eBiosciences, San Diego, unless otherwise noted) following standard procedures: anti-Osteopontin (Abcam), anti-Nestin (MAB353, Millipore), anti-Gr-1 (RB6-8C5), anti-Mac-1 (M1/70), anti-B220 (RA3-6B2), anti-Ter-119 (TER-119), anti-CD3 (145-2C11, BD Biosciences), anti-CD115 (AFS98), anti-CD150 (TC15-12F12.2, BD Biosciences), anti-CD31 (MEC13.3, Biolegend), anti-CD48 (HM48-1), and anti-CD41 (eBioMWReg30) antibodies. Images were acquired using Olympus Confocal Laser Scanning Biological Microscope FV1000 equipped with four lasers ranging from 405 to 635 nm. Images were processed with ImageJ software.

Dong L., Yu W.M., Zheng H., Loh M.L., Bunting S.T., Pauly M., Huang G., Zhou M., Broxmeyer H.E., Scadden D.T, & Qu C.K. (2016). Leukaemogenic effects of Ptpn11 activating mutations in the stem cell microenvironment. Nature, 539(7628), 304-308.

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