Mouse anti cd16 32 antibody

Manufactured by Cytek Biosciences

Sourced in United States

The mouse anti-CD16/32 antibody is a laboratory reagent used for flow cytometry analysis. It binds to the CD16 and CD32 receptors, which are expressed on the surface of various immune cells, such as macrophages, dendritic cells, and natural killer cells. This antibody can be used to detect and quantify the presence of these receptors on cell populations.

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Lab products found in correlation

Zombie yellow fixable viability kit, by BioLegend (1 mentions) Fluorescein isothiocyanate conjugated anti cd4, by Thermo Fisher Scientific (1 mentions) Cytexpert software, by Beckman Coulter (1 mentions) Lsrfortessa analyzer, by BD (1 mentions) Ter119, by BioLegend (1 mentions) Alexa fluor 700, by BioLegend (1 mentions) Cd135, by BioLegend (1 mentions) Ly6g 6c, by BioLegend (1 mentions) Cd11b, by BioLegend (1 mentions) Lsrfortessa cell analyzer, by BD (1 mentions) Donkey serum, by Merck Group (1 mentions) Prolong diamond antifade mountant, by Thermo Fisher Scientific (1 mentions)

Close spelling variants of this product

Anti-mouse CD16/32 antibody (2.4G2) Anti-mouse CD16/32 Anti-CD16/32 CD16/32

4 protocols using mouse anti cd16 32 antibody

Phenotypic Analysis of Murine Cervical Lymphocytes

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A single-cell suspension was prepared from the cervical lymph nodes removed from three mice. After lysis of red blood cells in 1 × RBC Lysis Buffer Solution (eBioscience, San Diego, CA, USA) and selection of live cells using the Zombie Yellow Fixable Viability Kit (BioLegend, San Diego, CA, USA), lymphocytes were incubated with mouse anti- CD16/32 antibody (Tonbo Biosciences, San Diego, CA, USA) followed by incubation with fluorescein allophycocyanin-conjugated mouse anti-CD8a (BD Biosciences, San Jose, CA, USA), phycoerythrin-Cy7-conjugated mouse anti-CD69 (BioLegend), BrilliantViolet785-conjugated anti-mouse CD3 (BioLegend), fluorescein isothiocyanate-conjugated anti-CD4 (eBioscience), or fluorophore-conjugated isotype controls (eBioscience). The cells were analyzed using CytoFLEX (V5-B5-R3 configuration, Beckman Coulter, Fullerton, CA, USA), and data were analyzed with CytExpert software (Beckman Coulter).

Uchihashi T., Nakahara H., Fukuhara H., Iwai M., Ito H., Sugauchi A., Tanaka M., Kogo M, & Todo T. (2021). Oncolytic herpes virus G47Δ injected into tongue cancer swiftly traffics in lymphatics and suppresses metastasis. Molecular Therapy Oncolytics, 22, 388-398.

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ALDH1 and CD44 Expression Analysis

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Cells were trypsinated at about 80% confluence and the cell number was counted. 20E4 cells were resuspended in 1 ml ALDEFUOR™ Assay buffer, and 5 ul of activated substrate was added into the cell suspension. Then, 0.5 ml of the mixture was transferred to another tube with 5 ul DEAB to inactivate the ALDH enzymatic reaction. Both the DEAB and test samples were incubated at 37°C for 45 min. For CD44 staining, cells were blocked with mouse anti-CD16/32 antibody (Tonbo Biosciences) for 10 minutes and then stained with APC conjugated CD44 antibody (Tonbo Biosciences) on ice for 30 minutes. ALDH+ cells and CD44 expression were then examined with BD LSR Fortessa Analyzer, and analyzed with FlowJo v10.0 (BD). The percentage of ALDH1+ population in test samples was determined with the same gate containing 0.1% positive cells in the corresponding DEAB sample.

Zhang W., Bado I.L., Hu J., Wan Y.W., Wu L., Wang H., Gao Y., Jeong H.H., Xu Z., Hao X., Lege B.M., Al-Ouran R., Li L., Li J., Yu L., Singh S., Lo H.C., Niu M., Liu J., Jiang W., Li Y., Wong S.T., Cheng C., Liu Z, & Zhang X.H. (2021). The bone microenvironment invigorates metastatic seeds for further dissemination. Cell, 184(9), 2471-2486.e20.

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Comprehensive Immune Cell Profiling by Flow Cytometry

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Bone marrow cells, thymocytes, splenocytes, or peripheral blood cells were isolated, and RBC lysis buffer was added to remove the RBCs. Cells were stained at a 1:200 dilution with 15 mouse fluorochrome-conjugated monoclonal antibodies specific for the following murine cell surface markers encompassing the major immune lineages: B220, CD19, IgM, IgD, CD3ε, CD4, CD5, CD11c, CD44, CD43, CD25, CD21, CD23, BP-1 (BD PharMingen), CD8α, CD11b, NK1.1 (Biolegend), F4/80, and CD62L (Tonbo Biosciences), and in the presence of anti-mouse CD16/32 antibody (Tonbo Biosciences) for 1 h at 4°C. After staining, cells were washed twice in PBS and analyzed by flow cytometry. To stain the hematopoietic progenitor compartment, bone marrow was isolated and stained with Alexa Fluor 700–conjugated lineage markers (CD3, Ly-6G/6C, CD11b, B220, and Ter-119 at a 1:50 dilution; Biolegend), c-Kit, Sca-1, CD16/32, CD34, IL-7Rα (BD PharMingen), and CD135 (Biolegend) at a 1:100 dilution for 1 h at 4°C. After staining, cells were washed twice in PBS and analyzed by flow cytometry. Data were acquired on an LSRFortessa cell analyzer (BD Bioscience) and analyzed with FlowJo software (Treestar).

Choi J.H., Zhong X., Zhang Z., Su L., McAlpine W., Misawa T., Liao T.C., Zhan X., Russell J., Ludwig S., Li X., Tang M., Anderton P., Moresco E.M, & Beutler B. (2020). Essential cell-extrinsic requirement for PDIA6 in lymphoid and myeloid development. The Journal of Experimental Medicine, 217(4), e20190006.

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Circulating Tumor Cell Isolation from Mice

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500μl blood were draw from the right ventricle of anesthetized NRG mice inoculated with mammary tumors or bone metastases after 6 weeks. Blood samples were immediately mixed with 8 ml of red blood cell lysis buffer and incubated on ice for 10 minutes. Samples were then centrifuged at 250 g for 10 minutes at 4°C and the supernatant was discarded. The same steps were repeated once to completely remove red blood cells. Cell pellets were then re-suspended with cold PBS and transferred to poly-L-lysine coated slides. The slides were placed in the 37°C incubator for 30 minutes, and fixed with 4% PFA for 10 minutes. Fixed cells were rinsed with PBS for three times, and permeated with 0.3% Triton-X 100 for 30 minutes at RT. Slides were then blocked with donkey serum (Sigma) and anti-mouse CD16/32 antibody (Tonbo Biosciences) for 2 hours and incubated with fluorescence conjugated primary antibodies at 4 °C overnight. The next day, slides were stained with DAPI and mounted with Prolong™ Diamond Antifade Mountant (Molecular Probe). Circulating tumor cells were identified and imaged by a CyteFinder® instrument (Rarecyte) with same exposure setting.

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