Apc anti mouse cd3 antibody

Manufactured by BioLegend

Sourced in United States

The APC anti-mouse CD3 antibody is a flow cytometry reagent that binds to the CD3 protein expressed on the surface of mouse T cells. It can be used to identify and enumerate T cells in mouse samples.

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Anti-CD3 (468 citations) CD3-APC (111 citations) Anti-CD3 antibody (77 citations) Anti-CD3-APC (66 citations) Anti-mouse CD3 (46 citations) APC anti-mouse CD3 (21 citations) Anti-mouse CD3 antibody (10 citations) APC-conjugated anti-mouse CD3 antibody (4 citations) APC-CD3 antibody (3 citations) APC/Cyanine7 anti-mouse CD3 antibody (2 citations)

7 protocols using apc anti mouse cd3 antibody

Evaluation of Morinda officinalis in Colitis

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Morinda officinalis (no. 20150603) was obtained from Shenzhen Lush Pharmaceutical Co., Ltd. Positive drug mesalazin enteric-coated tablets (no. 150308) was bought from Losan Pharma GmbH (Germany). DSS was purchased from MP Biomedicals (MW; 36,000–50,000; MP Biomedicals, Solon, OH, USA). The enzyme-linked immunosorbent assay (ELISA) kits for TNF-α, IL-6, and IL-17 was bought from Huamei (Cusabio Biotech Co. Ltd., China). All plastic materials were purchased from Falcon Labware (Becton-Dickinson, Franklin Lakes, NJ, USA). RPMI Medium 1640, fetal bovine serum (FBS), and phosphate buffer saline (PBS) were obtained from GIBCO Laboratories (Grand Island, NY, USA). Penicillin G/streptomycin, MTT, and dimethyl sulfoxide (DMSO) were purchased from Sigma (St. Louis, MO, USA). Apoptosis Assay Kit was purchased from Lianke Biology Inc. (Hangzhou, China). APC antimouse CD3 antibody, PE/Cy7 antimouse CD4 antibody, and APC antimouse CD8 antibody were purchased from BioLegend (USA). All other chemicals used were analytical grade and were supplied by the Beijing Chemical Agents Company (China).

Liang J., Liang J., Hao H., Lin H., Wang P., Wu Y., Jiang X., Fu C., Li Q., Ding P., Liu H., Xiong Q., Lai X., Zhou L., Chan S, & Hou S. (2017). The Extracts of Morinda officinalis and Its Hairy Roots Attenuate Dextran Sodium Sulfate-Induced Chronic Ulcerative Colitis in Mice by Regulating Inflammation and Lymphocyte Apoptosis. Frontiers in Immunology, 8, 905.

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Comprehensive T Cell Analysis in Murine Blood

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Blood was collected from mice (50 μL in each staining tube). Each tube received 1 × Lysis Buffer (Biolegend, San Diego, CA, USA) followed by incubation at 37 °C for 10 to 15 min. The cells were centrifuged at 300 × g for 5 min. The supernatant was removed and the cells were stained with peridinin chlorophyll protein complex (PerCP) anti-mouse CD45 antibody, APC anti-mouse CD3 antibody, PE anti-mouse CD8b antibody, FITC anti-mouse CD4 Antibody, True-Nuclear™ One Step Staining Mouse Treg Flow™ Kit (FOXP3 Alexa Fluor® 488/CD25 PE/CD4 PerCP), and FITC anti-mouse IgG1 antibody, PE anti-mouse IgG1 antibody, and APC anti-mouse IgG1 antibody (all from Biolegend, San Diego, CA, USA) as an isotype control. Samples were run on a BD Canto II flow cytometer. The percentage of proliferating T cells was analyzed using FlowJo version 10.

Bai X., Chen T., Li Y., Ge X., Qiu C., Gou H., Wei S., Liu T., Yang W., Yang L., Liang Y., Jia Z., Lv L, & Li T. (2023). PD-L1 expression levels in mesenchymal stromal cells predict their therapeutic values for autoimmune hepatitis. Stem Cell Research & Therapy, 14, 370.

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Multicolor Flow Cytometry Analysis of Immune Cells

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Primary cell suspensions from the thymus, spleen, or LNs or Raji cells with or without TRIB3 deletion were directly labeled. Fluorescently labeled antibodies against the following surface proteins were used for human/mouse cell staining: FITC anti-human CD19 antibody (Biolegend, 392508, 1:100), PE anti-human CD3 antibody (Biolegend, 981004, 1:100), APC anti-mouse CD3 antibody (Biolegend, 100236, 1:100), FITC anti-mouse/human CD45R/B220 antibody (Biolegend, 103205, 1:100), FITC anti-mouse Ki-67 antibody (Biolegend, 652410, 1:100), PE anti-mouse/human CD11b antibody (Biolegend, 101207, 1:100), FITC anti-human CD11b antibody (Biolegend, 301330,1:100). Data were acquired using a FACSCalibur or FACSCanto II flow cytometer (BD). FCS EXPRESS software was used for data analysis.

Li K., Wang F., Yang Z.N., Zhang T.T., Yuan Y.F., Zhao C.X., Yeerjiang Z., Cui B., Hua F., Lv X.X., Zhang X.W., Yu J.J., Liu S.S., Yu J.M., Shang S., Xiao Y, & Hu Z.W. (2020). TRIB3 promotes MYC-associated lymphoma development through suppression of UBE3B-mediated MYC degradation. Nature Communications, 11, 6316.

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Synthesis and Characterization of Cy5.5-Labeled Immune Checkpoint Inhibitor

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2-Chlorotrityl chloride resin (1.2 mmol/g) was purchased from Nankai University resin Co., Ltd. Fmoc-amino acids were obtained from GL Biochem (Shanghai, China). Cy5.5 NHS ester (non-sulfonated) purchased from APEXBIO. Alkali phosphatase (30 U/μL) was obtained from Takara (D2250, Dalian, China) Bio. Inc. Enhanced ATP Assay Kit (S0027) purchased from Beyotime. Both of anti-HMGB1 antibody (ab79823) and anti-carlreticulin antibody (ab92516) were purchased from Abcam. Calreticulin Rabbit mAb (Alexa Fluor 488 conjugate, #62304) was obtained from Cell Signaling Technology. HMGB1 ELISA Kit (OKEH00424) was purchased from AVIVA systems biology. InvivoMAb antimouse PD-L1 (B7-H1) was obtained from BioXcell. APC anti-mouse CD3 antibody (100235), FITC anti-mouse CD8a antibody (100705), APC anti-mouse IFN-γ (505810), PE anti-mouse CD80 (104707), FITC anti-mouse CD86 (105005), Alexa Fluor 647 anti-mouse CD11c (117314), ELISA MAX™ Deluxe Mouse IFN-γ, ELISA MAX™ Deluxe Mouse TNF-α and ELISA MAX™ Deluxe Mouse IL-6 were purchased from Biolegend. Collagenase IV and DNase I were both purchased from Solarbio. Commercially available reagents and solvents were used without further purification, unless noted otherwise.

Zheng D., Liu J., Xie L., Wang Y., Ding Y., Peng R., Cui M., Wang L., Zhang Y., Zhang C, & Yang Z. (2021). Enzyme-instructed and mitochondria-targeting peptide self-assembly to efficiently induce immunogenic cell death. Acta Pharmaceutica Sinica. B, 12(6), 2740-2750.

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Comprehensive Flow Cytometry Antibody Panel

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The following flow cytometry antibodies were purchased from BioLegend company (San Diego, USA): CD16/32 (clone: 93), BV510 anti-mouse CD45 antibody (clone: 30-F11), BV711 anti-mouse CD8a antibody (clone: 53-6.7), APC anti-mouse CD3 antibody (clone: 17A2), BV421 anti-mouse Foxp3 antibody (clone: MF-14), PE anti-mouse CD206 antibody (clone: C068C2), BV605 anti-mouse CD11b antibody (clone: M1/70), PE/Cy7 anti-mouse F4/80 antibody (clone: QA17A29), BV650 anti-mouse CD25 antibody (clone: PC61), Percp/Cy5.5 anti-mouse Gr-1 (Ly-6G/Ly-6C) antibody (clone: RB6-8C5), APC/Cy7 anti-mouse CD86 antibody (clone: GL-1), FITC anti-mouse CD80 antibody (clone: 16-10A1), PE/Cy5 anti-mouse CD19 (clone: 6D5), and Alexa fluor 700 anti-mouse MHC class II antibody (clone: M5/114.15.2). Flow cytometry antibodies for BUV563 anti-mouse CD4 (clone: RM4-5) was purchased from BD Company (New York, USA).

Cao Z., Li D., Zhao L., Liu M., Ma P., Luo Y, & Yang X. (2022). Bioorthogonal in situ assembly of nanomedicines as drug depots for extracellular drug delivery. Nature Communications, 13, 2038.

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Quantifying Antigen-Specific T-Cell Responses

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The antigen-specific CD4⁺ and CD8⁺ T-lymphocyte responses to each immunized group were measured through an intracellular cytokine staining (ICS) assay. Briefly, 1 × 10⁶ splenocytes were seeded in a 6-well plate and treated with M. hyopneumoniae protein at a concentration of 10 μg/mL. Cells were labeled with APC anti-mouse CD3 antibody (Biolegend, San Diego, CA, USA), PE anti-mouse CD4 antibody (Biolegend), and FITC anti-mouse CD8a antibody (Biolegend) (PBS:CD3:CD4:CD8a = 377:10:5:8) at 4 °C for 25 min, away from light. After being washed twice with cold PBS buffer, cells were resuspended in PBS and analyzed using Accuri C6 Plus Flow Cytometry (BD Biosciences, Franklin Lakes, NJ, USA).

Liu W., Jiang P., Song T., Yang K., Yuan F., Gao T., Liu Z., Li C., Guo R., Xiao S., Tian Y, & Zhou D. (2023). A Recombinant Chimera Vaccine Composed of LTB and Mycoplasma hyopneumoniae Antigens P97R1, mhp390 and P46 Elicits Cellular Immunologic Response in Mice. Vaccines, 11(8), 1291.

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Flow Cytometry Analysis of T Cell Infiltration

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T cell infiltration was analyzed with flow cytometry. Tumor tissues from mice with different treatments were collected and digested into single cell suspension. Then cell suspension was incubated with APC anti-mouse CD3⁺ antibody (Biolegend) and PE anti-mouse CD8⁺ antibody (Biolegend) and analyzed with flow cytometry.

Cai S., Chen Z., Wang Y., Wang M., Wu J., Tong Y., Chen L., Lu C, & Yang H. (2021). Reducing PD-L1 expression with a self-assembled nanodrug: an alternative to PD-L1 antibody for enhanced chemo-immunotherapy. Theranostics, 11(4), 1970-1981.

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