Pacific blue anti mouse human cd11b antibody

Manufactured by BioLegend

The Pacific Blue™ anti-mouse/human CD11b Antibody is a laboratory tool used for the detection and analysis of CD11b, a cell surface glycoprotein expressed on myeloid cells, including monocytes, macrophages, and granulocytes. This antibody is conjugated with the Pacific Blue™ fluorescent dye, allowing for the visualization and quantification of CD11b-expressing cells through flow cytometry or other immunoassay techniques.

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3 protocols using pacific blue anti mouse human cd11b antibody

Radiolabeling and Immune Profiling

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All chemical reagents, purchased from Meryer Chemical (China), Zhengzhou Alfa (China), J&K (China), Energy Chemical (China), Xi’an Ruixi Biological Technology Co., Ltd (China) and Macrocyclics, Inc. Cell counting kit-8 (CCK-8) was purchased from Biyuntian Biotechnology Institute. Annexin V-FITC/PI Apoptosis Detection Kit was purchased from Yeasen Biotechnology (Shanghai) Co., Ltd. ⁸⁹Zr was produced on an onsite cyclotron by the ⁸⁹Y(p,n)⁸⁹Zr reaction and purified to yield ⁸⁹Zr(C₂O₄)₂ solution. The cell Apoptosis 7-AAD Detection Kit was purchased from KeyGen Biotech Co., Ltd (Nanjing, China). FITC anti-mouse CD45 Antibody (#103107, 1:200), Pacific Blue™ anti-mouse/human CD11b Antibody (#101223, 1:100), APC/Cyanine7 anti-mouse F4/80 Antibody (#123117, 1:100), PE/Cyanine7 anti-mouse CD86 Antibody (#105013, 1:200), PE anti-mouse CD206 (MMR) Antibody (#141705, 1:100), PE-conjugated anti-mouse CD3 Antibody (#100205, 1:100), FITC-conjugated anti-mouse CD4 (#100406, 1:200), APC-conjugated anti-mouse CD8a Antibody (#100712, 1:200), Purified anti-HMGB1 Antibody (#651401, 1:100) were purchased from Biolegend. Mouse TNF-α, IL-6, IL-12p70 ELISA Kits and eFluor-450-conjugated anti-mouse Foxp3 antibody (#48-5773-82. 1:200) were obtained from Invitrogen. Alexa Fluor® 488 Anti-Calreticulin antibody [EPR3924] (ab196158, 1:100) was bought from Abcam.

Shi Y., Guo Z., Fu Q., Shen X., Zhang Z., Sun W., Wang J., Sun J., Zhang Z., Liu T., Gu Z, & Liu Z. (2023). Localized nuclear reaction breaks boron drug capsules loaded with immune adjuvants for cancer immunotherapy. Nature Communications, 14, 1884.

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Assessing Microglial Responses to Amyloid-Beta

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Mouse hippocampal and PFC extracts were prepared as reported previously⁵⁹. The accumulation of human Aβ_1–40 and Aβ_1–42 was quantified by ELISA (catalog nos. KHB3442 (Amyloid beta 42 Human ELISA Kit) and KHB3482 (Amyloid beta 40 Human ELISA Kit); Thermo Fisher Scientific). Microglia were isolated from the cortex and hippocampus of WT and APP/PS1 mice using fluorescence activated cell sorting (FACS) method and CD11b⁺and CD45^low cells were isolated. The antibodies used were Pacific Blue anti-mouse/human CD11b antibody (catalog no. 101224; BioLegend) and Alexa Fluor 647 anti-mouse CD45 antibody (catalog no. 103124; BioLegend). Cultured microglia were treated with Pink1 siRNA (E-004030–00-0005; Dharmacon) for 72 h and then treated with 50 μM UA for 24 h. The supernatants were analyzed by ELISA. The kits used for the cytokines were obtained from R&D Systems, including the mouse TNF-alpha (catalog no. MTA00B), IL-6 (catalog no. M6000B), and IL-10 (catalog no. M1000B) Quantikine ELISA Kits.

Fang E.F., Hou Y., Palikaras K., Adriaanse B.A., Kerr J.S., Yang B., Lautrup S., Hasan-Olive M.M., Caponio D., Dan X., Rocktäschel P., Croteau D.L., Akbari M., Greig N.H., Fladby T., Nilsen H., Cader M.Z., Mattson M.P., Tavernarakis N, & Bohr V.A. (2019). Mitophagy inhibits amyloid-β and tau pathology and reverses cognitive deficits in models of Alzheimer’s disease. Nature neuroscience, 22(3), 401-412.

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Detailed Immunoblot Antibody Protocol

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Antibodies used for immunoblot are listed here based on where they are acquired. 1. From Cell Signaling Technology (Danvers, MA): anti-pS6 (4858); anti-pS6K (9204); anti-S6 (2317); anti-p4E-BP1 (2855); anti-4E-BP1(9452); anti-mTOR (2983); anti-pAkt (Ser473) (4060); anti-pAkt (Thr308) (13038); anti-Akt (2920). 2. From BioRad (Hercules, California): anti-CD68 (MCA19571); 3. From Biolegend (San Diego, California): Alexa fluoro-labeled anti-CD 206 (141709); APC anti-mouse Ly-6G Antibody (127614); PE anti-mouse F4/80 Antibody (123110); Pacific Blue™ anti-mouse/human CD11b Antibody (101224); PE/Cy7 anti-mouse CD45 (103114); Brilliant Violet 711™ anti-mouse Ly-6C Antibody (128037); PerCP/Cy5.5 anti-mouse CD206 (MMR) Antibody (141716). 4. From Santa Cruz Biotechnology (Santa Cruz, CA): mouse anti-GAPDH antibody; anti-LAMP1 (sc-20011). 5. From R&D Systems (Minneapolis, MN): goat anti-CD31 (AF3628). 6. From Cedarlane (Burlington, Ontario, Canada): anti-Mouse Mac-3 (CL8993AP).

Chen G., Phan V., Lou X, & Cao D.J. (2021). The mechanistic target of rapamycin complex 1 critically regulates the function of mononuclear phagocytes and promotes cardiac remodeling in acute ischemia. Journal of molecular and cellular cardiology, 159, 62-79.

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