Mice were treated i.p. with 2.5 mg/kg poly I:C HMW (InvivoGen) every 2 days for 12 consecutive days. The weight of the mice was recorded daily. Mice were euthanized and analyzed on day 13. For blockading, CD22 mice were treated with an anti-CD22 antibody every 2 days for 12 consecutive days (InVivoMAb anti–mouse CD22; clone Cy34.1, Bio X Cell). A 20 μg dose of antibody in PBS was given in a final volume of 100 μL, or an isotype control mAb (clone MOPC-21; Bio X Cell) was given. For the CD20 depletion experiment, the mice were administered i.p. 10 μg anti-CD20 mAB (clone MB20-11; Bio X Cell) or an isotype control mAb (clone MOPC-21; Bio X Cell) every 2 days, 6 days before the start of the experiment.
Clone mopc 21
Manufactured by BioXCell
Sourced in United States
The Clone MOPC-21 is a laboratory equipment product for cell culture research. It is a monoclonal antibody that can be used for various applications in cell biology. The core function of this product is to provide a specific binding reagent for target antigens of interest.
Automatically generated - may contain errors
Lab products found in correlation
Clone 1d11, by BioXCell (3 mentions)
Anti h 2kb siinfekl antibody clone 25 d1, by BioXCell (2 mentions)
Dynabeads untouched mouse cd8 cells, by Thermo Fisher Scientific (2 mentions)
2 mercaptoehanol, by Thermo Fisher Scientific (2 mentions)
Recombinant murine il 2, by Thermo Fisher Scientific (2 mentions)
Carboxyfluorescein succinimidyl ester (cfse), by BioLegend (2 mentions)
Celltiter glo, by Promega (2 mentions)
Clone ltf 2, by BioXCell (2 mentions)
Anti tnfα clone xt3, by BioXCell (1 mentions)
Celltrace yellow, by Thermo Fisher Scientific (1 mentions)
Anti mouse cd47 mab clone miap410, by BioXCell (1 mentions)
Iscove modified dulbecco media (imdm), by Thermo Fisher Scientific (1 mentions)
Murine m csf, by Thermo Fisher Scientific (1 mentions)
Cd16 apc cy7, by BD (1 mentions)
Clone fn50, by BD (1 mentions)
Interleukin 2 (il 2), by Thermo Fisher Scientific (1 mentions)
Clone cd28, by BD (1 mentions)
Efluor 670, by BD (1 mentions)
Clone rpa t4, by BD (1 mentions)
Clone okt 3, by BioXCell (1 mentions)
21 protocols using clone mopc 21
1
Poly I:C-Induced Immune Response in Mice
2
Adoptive T Cell Transfer in Typhus Infection
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Mice were infected subcutaneously (s.c.) into the tail base with 2×106 sfu R. typhi in 50 μl PBS. Either 1×106 purified CD8+ or CD4+ T cells from naïve BALB/c, BALB/c IFNγ-/- or BALB/c Perforin-/- mice were adoptively transferred 1 day prior to infection. TNFα was neutralized by intraperitoneal application of 500 μg anti-TNFα (clone XT3.11, BioXCell, West Lebanon, US) in 200 μl PBS. Control mice received the same amount of isotype antibody (clone HRPN, BioXCell, West Lebanon, US). Treatment was performed every three days beginning on day 3 post infection. For the neutralization of IL-17A, anti-IL-17A (clone 13F3) was used (BioXCell, West Lebanon, US). 500 μg of the antibody were injected i.p. in 200 μl PBS every 2 days starting on day 2 post infection. Control mice received the same amount of isotype antibody (clone MOPC-21, BioXCell, West Lebanon, US).
3
PDAC Organoids Potentiate CD8+ T Cell Killing
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Mouse PDAC cells stably expressing OVA and carrying doxycycline (Dox)-inducible mTurquoise2-Atg4BC74A were grown as organoids, treated with or without Dox (1 μg/mL) for 96 hrs. Organoids were dissociated into single cells, which were then incubated with either anti-H-2Kb-SIINFEKL antibody (clone 25-D1.16, BioXCell, BE0207) or isotype control (clone MOPC-21, BioXCell, BE0083) at 100 μg/mL for 30 min at 4 °C. Total splenocytes were harvested from OT-I mice and CD8+ T cells were enriched using Dynabeads® Untouched™ Mouse CD8 Cells (Invitrogen, 11417D) following manufacturer’s instructions. Isolated CD8+ T cells were labelled with 10 μM CFSE (BioLegend) for 10 min at RT in the dark, washed 3x with RPMI-1640 supplemented with 10% FBS. Ten thousand PDAC cells and forty thousand CD8+ cells were seeded in 96-well plates and cultured in 100 μL 50% DMEM and 50% RPMI-1640 supplemented with 10% FBS, 10 ng/mL recombinant murine IL-2 (Peprotech), 27.5 μM 2-Mercaptoehanol (Gibco), and 100 μg/mL of respective antibodies. After 48 hrs, CD8+ T cells were harvested and stained with anti-CD8a antibody (AF647, clone 53–6.7, BioLegend) and DAPI, and proliferation was analyzed by CFSE dilution using flow cytometry. After removal of CD8+ T cells, the viability of remaining PDAC cells were measured by CellTiter-Glo (Promega).
4
Phagocytosis Assay of JAK2 Mutant Erythrocytes
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Spleen cells were harvested from WT and JAK2 mutant mice and macrophages were generated by culturing the cells in IMDM (Invitrogen) with 10% FBS and murine M-CSF (PeproTech) for 6 days. Macrophages were detached and plated in a 24-well plate with 2.5 × 105 cells per well with 1 ml medium 1 day prior to the assay. The following day, RBCs from PV mice were incubated with CellTrace™ Yellow (Invitrogen) according to the manufacturers protocol. The RBCs were added to the macrophages at an effector to target ratio of 1:10 and the anti-mouse CD47 mAb (clone MIAP410; BioXCell) or a mouse IgG1 isotype control (clone MOPC-21; BioXCell) were added at a concentration of 5 μg/ml. The macrophages and RBCs were co-incubated for 2 h, then washed, stained, and analyzed. Phagocytosis of RBCs was defined by flow cytometry as the percentage of CellTrace™ Yellow-labeled cells out of F4/80+Aqua- cells.
5
Activated T Cell Phenotyping by Flow Cytometry
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Flow cytometry was performed on a FACS LSR II or Fortessa (BD Biosciences). For lymphocyte activation studies, soluble anti-CD3 (500 ng/ml; clone OKT3, Bio X Cell) ± anti-CD28 (1 μg/ml; clone CD28.2, BD Biosciences) or IL-2 (PeproTech) was added to media for 2 days. Proliferation was measured by CFSE (eFluor 670, BD Biosciences) after 3 days. EVs were added every 24 hours (5 μg/ml). PD1 blockade was achieved by adding anti-PD1 (10 μg/ml; EH12) or immunoglobulin G control (clone MOPC-21, Bio X Cell). Cells were gated by FSC (forward scatter)/SCC (side scatter) while excluding duplets by FSC-A/FSC-H. Subsequently, CD3+CD56− T cells were gated with further classification of CD4+ and CD8+ T cells. Finally, CD4+ and CD8+ T cells were measured for their CD69, CD25, PD1, and TIM3 expression (seen as a downloadable figure). Cells were stained with the following antibodies: CD3/PE-Cy5.5 (1:100; clone SK7, eBioscience), CD4/FITC (1:100; clone RPA-T4, BD Biosciences), CD8/AmCyan (1:10; clone SK1, BD Biosciences), CD16/APC-Cy7 (1:100, BD Biosciences), CD25/PE-Cy7 (1:10; clone BC96, eBioscience), CD56/V450 (1:30; clone B159, BD Biosciences), CD69/APC (1:10; clone FN50, BD Biosciences), TIM3/APC-Cy7 (1:100, clone F38-2E2, BD Biosciences), and PD1/PE (1:100; clone J105, eBioscience).
6
PDAC Organoids Potentiate CD8+ T Cell Killing
7
Anti-IFNAR1 Antibody Modulates Mycobacterial Burden
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In each group, 12 mice C57BL/6 mice were treated twice with 500 μg of anti-IFNAR1 antibody (clone MAR1-5A3; Biolegend San Diego, CA, USA) or mouse IgG1 (500 μg) isotype control (clone MOPC21; Bio X Cell) one day prior to M. bovis infection and at 6 weeks post infection through intraperitoneal (i.p) injection. For the enumeration of total viable bacilli, lung and spleen tissues were lysed with small ceramic beads in phosphate buffered saline (PBS), in a tissue homogenizer apparatus (WKT technology) in accordance with the guidelines of manufacturer. An appropriate tenfold serial dilution was prepared in PBS. The dilutions were separately plated in triplicates on Middlebrook 7H11 agar supplemented with ampicillin (10 μg/ml) and sodium pyrovate (2-4 mg/liter). After 2–3 weeks of incubation at 37 °C, M. bovis colonies were counted [21 (link)].
8
In vivo Murine Klebsiella pneumoniae Infection
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For infection experiments, mice were i.n. challenged with 5 × 104 CFUs of K. pneumoniae in 30 μl of sterile PBS. For in vivo antibody-mediated blocking experiments, mice were intraperitoneally (i.p.) injected with the following antibodies 24–48 h before infection: αMR1 (150 μg/mouse; clone 26.5; Biolegend, Mouse IgG2a), αIFNAR1 (200 μg/mouse; clone MAR1-5A3; BioXCell, Mouse IgG1), αSiglecH (150 μg/mouse; clone 440c; BioXCell, Rat IgG2b), or their respective isotype controls: Mouse IgG2a (150 μg/mouse; clone MOPC-173; Biolegend), Mouse IgG1 (200 μg/mouse; clone MOPC-21; BioXCell), and Rat IgG2b (150 μg/mouse; clone LTF-2; BioXCell). When indicated, MAIT cell expansion in vivo was performed through a repeated i.n. inoculation (3×) of 5-OP-RU (100 μM) and LPS (17.4 μg/mouse; Invivogen).
Bacterial loads were determined by counting CFUs after plating 100-fold dilution series of tissue homogenates obtained from bacteria-infected mice. Colonies were counted at 24 h.
Bacterial loads were determined by counting CFUs after plating 100-fold dilution series of tissue homogenates obtained from bacteria-infected mice. Colonies were counted at 24 h.
9
Anti-IFNAR1 Modulates Staphylococcus Infection
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Mice were i.p. inoculated with 2.5 mg anti-mouse IFNAR1 neutralizing mAb (clone MAR1-5A3, BioXcell) or 2.5 mg IgG isotype control (Clone MOPC-21, BioXcell). Twenty-four hours later, the mice were anesthetized with pentobarbital sodium and i.v. challenged with 2 × 108 CFU of Staphylococcus suspended in 200 μL PBS. Mortality was monitored.
10
Anti-CSF1R Treatment in Murine Cancer
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Mice were treated intraperitoneally (i.p.) with 30 mg/kg of either mouse-hamster chimeric IgG1 anti-mouse CSF1R, clone 2G2 (Roche) (Ries et al., 2014 (link)); or a control mouse IgG1 (clone MOPC-21; BioXCell), both dissolved in PBS (Dulbecco’s PBS, Bioconcept). Treatments were initiated 4 days after cancer cell inoculation. All mice received two (MC38 model) or three (E0771 model) doses of the drugs, once per week.
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