Mopc 173

Manufactured by BioLegend

Sourced in United States

MOPC-173 is a monoclonal antibody that recognizes the mouse immunoglobulin kappa light chain. It is commonly used in research applications as a control or isotype-matched antibody.

Automatically generated - may contain errors

Lab products found in correlation

Close spelling variants of this product

Clone MOPC-173 (9 citations) IgG2a (clone MOPC-173, (3 citations) Mouse IgG2a isotype control (Clone MOPC-173; (2 citations) PE mouse IgG2a kappa isotype control (clone MOPC-173 (2 citations) Antibody (clone MOPC-173, (2 citations) Mouse IgG2a-APC (clone MOPC-173, (2 citations) IgG2bκ (clone MOPC‐173; (2 citations) APC mouse IgG2a kappa isotype control (clone MOPC-173 (2 citations) IgG2 isotype control (clone MOPC-173, (2 citations) Isotype control antibody (clone MOPC-173, (2 citations)

16 protocols using mopc 173

Comprehensive Immunological Profiling of PD-L1 Expression

Check if the same lab product or an alternative is used in the 5 most similar protocols

Immunofluorescence: mouse α-EEA1 (1G11, eBioscience), mouse α-PD-L1 (MIH1, eBioscience), rabbit α-PD-L1 (EPFR19759, Abcam), mouse α-LAMP1 (H4A4, eBioscience), mouse α-Calnexin (AF18, Abcam), mouse α-GM130 (169276, Abcam), mouse α-TGN46 (2F7.1, Abcam), mouse α-Rab11 (47/Rab11, BD Transduction Laboratories), mouse α-TfR (236-15375, Thermo-Fisher), rabbit α-CMTM6 (HPA026980, Sigma-Aldrich).
Flow Cytometry: FITC mouse IgG2a κ isotype control (MOPC-173, Biolegend), APC mouse IgG2b κ isotype control (MPC-11, Biolegend), APC mouse IgG1 κ isotype control (MOPC-21), APC mouse α-PD-L1 (29E.2A3, Biolegend), APC mouse α-PD-L1 (MIH1, eBioscience), Alexa Fluor 488 rabbit α-PD-L1 (Abcam), APC mouse α-HLA-ABC (W6/32), Alexa Fluor 488 mouse α-HLA-ABC (W6/32, Biolegend), FITC mouse α-CD8 (HIT8a, BD Pharmingen), PE-Cy7 mouse α-TNFα (MAb11, eBioscience), APC mouse α-PD-1 (J105, eBioscience), PerCp-Cy5.5 mouse α-Perforin (B-D48, Biolegend).
Immunoprecipitation/Immunoblotting: goat anti-PDL1 (AF156, R&D Systems), rabbit α-PD-L1 (E1L3N, Cell Signalling Technology), mouse α-PD-L1 (405.9A11, Cell Signalling Technology), mouse α-PD-L1 (MIH1, eBioScience), rabbit α-CMTM6 (Sigma-Aldrich, HPA026980).

Burr M.L., Sparbier C.E., Chan Y.C., Williamson J.C., Woods K., Beavis P.A., Lam E.Y., Henderson M.A., Bell C.C., Stolzenburg S., Gilan O., Bloor S., Noori T., Morgens D.W., Bassik M.C., Neeson P.J., Behren A., Darcy P.K., Dawson S.J., Voskoboinik I., Trapani J.A., Cebon J., Lehner P.J, & Dawson M.A. (2017). CMTM6 maintains the expression of PD-L1 and regulates anti-tumour immunity. Nature, 549(7670), 101-105.

+ Open protocol

+ Expand

TIGIT and TIM-3 Blockade Enhances CD8+ T-cell Function

Check if the same lab product or an alternative is used in the 5 most similar protocols

We selected cryopreserved PBMCs from S1 (n=10) and S2 (n=10). Samples were previously characterized by the expression of TIGIT and TIM-3 on total CD8+ T-cells. PBMCs were thawed and rested for four hours at 37 °C in a 5% CO₂ incubator. Next, cells were incubated under RPMI complemented medium 10% FBS with 1 μl/ml of anti-CD28/CD49d and 1 μl/ml of Monensin A overnight at 37 °C in a 5% CO₂. PBMCs are divided in the following conditions; (1) unstimulated, (2) SEB (1 μg/ml, Sigma-Aldrich) and (3) HIV-1-Gag peptide pool (2 μg/peptide/ml) in the absence or presence of αTIGIT and/or αTIM-3, and its respective isotype antibodies. For the single blockade of TIGIT (αTIGIT), we included Ultra-LEAF purified anti-human TIGIT antibody (10 μg/ml, clone A15153G, Biolegend) or its control isotype Ultra-LEAF purified mouse IgG2a antibody (10 μg/ml, MOPC-173, Biolegend). For single TIM-3 blockade (αTIM-3), we used Ultra-LEAF purified anti-human TIM-3 antibody (10 μg/ml, clone F38-2E2, Biolegend) or its respective isotype Ultra-LEAF purified mouse IgG1 antibody (10 μg/ml, MOPC-21, Biolegend). Finally, we included αTIGIT+αTIM-3 or their respective IgG2 + IgG1 isotypes for a combinational blockade. The next day, PBMCs were surface and intracellularly stained with the panel of antibodies and the methodology described in the section above.

Blanch-Lombarte O., Ouchi D., Jimenez-Moyano E., Carabelli J., Marin M.A., Peña R., Pelletier A., Talla A., Sharma A., Dalmau J., Santos J.R., Sékaly R.P., Clotet B, & Prado J.G. (2023). Selective loss of CD107a TIGIT+ memory HIV-1-specific CD8+ T cells in PLWH over a decade of ART. eLife, 12, e83737.

+ Open protocol

+ Expand

Blocking Leukocyte Migration Factors

Check if the same lab product or an alternative is used in the 5 most similar protocols

The following mAbs were used: Mel-14 (rat IgG2a, anti–murine L-selectin ((45 (link)), Bioexcell) and PS/2 (rat IgG2b, anti–murine α4) mAbs were stored at −70°C in endotoxin-free saline at 1 mg/ml. P-selectin and E-selectin were blocked with RMP-1 and ultra-RME-1/CD62E (Biolegend) respectively. The integrin α4β7 (LPAM) was blocked with DATK32 (Biolegend), CCR9 with blocking antibodies (9B1, Biolegend) and CXCR3 with blocking antibodies (CXCR3–173, Biolegend). For inhibition studies, 100 μg of antibody was directly injected i.v. before cell transfer (app. 1–2 h) or co-injected with pDC suspension. Administration of the S1PR inhibitor FTY720 (1 mg/kg) or PBS was performed as indicated or by daily i.p. injections.
For long term blockade mice were fed with SFD (D12450J, research diets) or HFD (D12492, research diets) for 1 week and injected daily i.p. with 100 μg of mAb against P-selectin (Biolegend) or its isotype control (IgG2a, MOPC-173). Mice were weighed daily and the weight of VAT was analyzed on day 7. glucose tolerance was tested (IPGTT) by injecting 1 mg/kg glucose (Gibco) i.p. into 5 h starved mice. The level of blood glucose was measured at 10 min intervals until 90 min after glucose injection.

Stutte S., Ishikawa-Ankerhold H., Lynch L., Eickhoff S., Nasiscionyte S., Guo C., van den Heuvel D., Setzensack D., Colonna M., Maier-Begandt D., Weckbach L., Brocker T., Schulz C., Walzog B, & von Andrian U. (2022). High-Fat Diet Rapidly Modifies Trafficking, Phenotype, and Function of Plasmacytoid Dendritic Cells in Adipose Tissue. The Journal of Immunology Author Choice, 208(6), 1445-1455.

+ Open protocol

+ Expand

Monitoring Glycolytic Shift in T-Cell Activation

Check if the same lab product or an alternative is used in the 5 most similar protocols

To monitor the glycolytic switch upon activation, CD4⁺ and CD8⁺ T-cells were resuspended in serum-free XF Assay media supplemented with 11.1 mM glucose and 2 mM l-glutamine (Sigma). ECAR and OCR were measured simultaneously throughout the experiment, i.e., 1 h before activation and 4 h after. T-cells were activated via the multi-injection port with anti-CD3 (0.2 µg/ml; HIT3a, BioLegend) and CD28 (20 µg/ml; CD28.2, BioLegend). A final injection of 2-DG (100 mM) was used to immediately arrest glycolysis. Isotype controls, mIgG2a κ (0.2 µg/ml; MOPC-173, BioLegend) and mIgG1 κ (20 µg/ml; MOPC-21; BioLegend) were used. The OCR/ECAR ratio was calculated by dividing the eight corresponding OCR and ECAR measurements pre- (dotted boxes) or post- (dashed boxes) antibody injection. Fold ECAR change was calculated by dividing the single point post antibody injection by the single point pre antibody injection. Peptide stimulation relied on the cross presentation of specific peptides by corresponding T-cell clones.

Jones N., Cronin J.G., Dolton G., Panetti S., Schauenburg A.J., Galloway S.A., Sewell A.K., Cole D.K., Thornton C.A, & Francis N.J. (2017). Metabolic Adaptation of Human CD4+ and CD8+ T-Cells to T-Cell Receptor-Mediated Stimulation. Frontiers in Immunology, 8, 1516.

+ Open protocol

+ Expand

Isolating and Infecting Primary Macrophages with H. pylori

Check if the same lab product or an alternative is used in the 5 most similar protocols

Primary macrophages cells were isolated from PBMC by adhesion to tissue culture plastic plates as described previously (21 (link)). Macrophages were >90% pure as determined by staining with CD14 and CD68. Isolated primary macrophages were harvested using a rubber policeman and were then infected with H. pylori at MOI of 20. For blocking experiments, H. pylori-infected macrophages were pre-treated with anti-MR-1 mAb (10 μg/ml, clone 26.5, Biolegend) or their matched isotype control mouse IgG2a (10 μg/ml, MOPC-173, Biolegend) before coculture with autologous effector cells.

Booth J.S., Salerno-Goncalves R., Blanchard T.G., Patil S.A., Kader H.A., Safta A.M., Morningstar L.M., Czinn S.J., Greenwald B.D, & Sztein M.B. (2015). Mucosal-Associated Invariant T Cells in the Human Gastric Mucosa and Blood: Role in Helicobacter pylori Infection. Frontiers in Immunology, 6, 466.

+ Open protocol

+ Expand

MAIT Cell Activation by Bacterial Ligands

Check if the same lab product or an alternative is used in the 5 most similar protocols

For co-cultures, C1R.MR1 and Jurkat.MAIT cells (at a ratio of 2:1) were incubated with supernatant from an overnight bacterial culture grown in LB. For ligand experiments, RL-6,7-DiMe (Toronto Research Chemicals) or 10 μM methylglyoxal with 5-A-RU^{26 (link)} was added directly to C1R.MR1 cells. Cells were then incubated for 18–20 h.
For PBMCs, cells were infected with live S. Paratyphi A strain NVGH308 or E. coli strain DH5α from frozen mid-log phase stocks. After 1 h, 100 μg/mL gentamicin was added and samples left for 16–18 h. For intracellular staining, 5 μg/mL brefeldin A solution was then added and incubated for 6 h. Intracellular Fixation & Permeabilization Buffer (eBioscience) was used for intracellular staining.
For MR1 blocking experiments, 10 μg/mL anti-MR1 antibody (clone 26.5, Biolegend) or mouse IgG2a isotype control (MOPC-173, Biolegend) were used. For IL-12 blocking experiments, LEAF™ purified anti-human IL12/23 p40 antibody (C11.5, Biolegend) and LEAF™ purified mouse IgG1 isotype control (MG1-45, Biolegend) were used. Antibodies were pre-incubated for 1 h with C1R.MR1 cells or PBMCs prior to addition of bacteria, supernatant, or ligand.

Howson L.J., Napolitani G., Shepherd D., Ghadbane H., Kurupati P., Preciado-Llanes L., Rei M., Dobinson H.C., Gibani M.M., Teng K.W., Newell E.W., Veerapen N., Besra G.S., Pollard A.J, & Cerundolo V. (2018). MAIT cell clonal expansion and TCR repertoire shaping in human volunteers challenged with Salmonella Paratyphi A. Nature Communications, 9, 253.

+ Open protocol

+ Expand

Flow cytometric analysis of immune cells

Check if the same lab product or an alternative is used in the 5 most similar protocols

For flow cytometric staining, antibodies against the following were used (Clone name, dilution, manufacturer and catalog number in brackets): CD3ε (145-2C11, 1:300–400, Biolegend #100312/100306), TCRβ (H57-597, 1:400, Biolegend #109222), CD4 (GK1.5, 1:300 Biolegend #100414), NKp46 (29A1.4, 1:50, Biolegend #137604), CD49b (HMα2, 1:200 Biolegend #103517), CD45 (30-F11, 1:500, Biolegend #103140), H2-K^b/D^b (28-8-6, 1:100, Biolegend #114606/114607), Qa-2 (695H1-9-9, 1:100, Biolegend #121709), Qa-1b (6A8.6F10.1A6, 1:10, Miltenyi Biotec #130-104-220), CD16/32 (93, 0.5 μg/10⁶ cells, Biolegend #101302), IL-17A (TC11-18H10.1, 1:125, Biolegend 506904), IFNγ (XMG1.2, 1:200, Biolegend #505809/eBioscience #17-7311-82), Mouse IgG2a, κ Isotype Ctrl (MOPC-173, 1:100, Biolegend #400207/400211), Mouse IgG1, κ Isotype Ctrl (MOPC-21, 1:66.7, Biolegend #400119), CD45.1 (A20, 1:300, Biolegend #110729), CD45.2 (104, 1:300, Biolegend #109806). For T-cell stimulation, antibodies against CD3ε (145-2C11, 3 μg/ml, eBioscience #16-0031-86) and CD28 (37.51, 1 μg/ml, eBioscience #16-0281-85) were used. For NK depletion assays, antibodies against NK1.1 (PK136, BioXCell #BP0036) or Isotype Control (C1.18.4, BioXCell #BP0085) were used.

Kang Y.H., Biswas A., Field M, & Snapper S.B. (2019). STAT1 signaling shields T cells from NK cell-mediated cytotoxicity. Nature Communications, 10, 912.

+ Open protocol

+ Expand

Evaluating HLA-II Expression in Ba/F3 Cells

Check if the same lab product or an alternative is used in the 5 most similar protocols

Ten μg/mL of DONQ52, control Ab (anti-KLH antibody, clone IC17-SG181), DQN0139, SPV-L3 (anti-pan HLA-DQ antibody, Beckman Coulter), Tu39 (anti-HLA-DR, DP, DQ antibody, BioLegend), or MOPC-173 (mouse IgG2a isotype antibody, BioLegend) was incubated with 1 × 10⁵ cells/100 μL/well Ba/F3 cells expressing HLA II in FACS buffer for 30 min at room temperature, followed by antibody detection with ×50 diluted Goat F(ab′)2 anti-Human IgG, Mouse ads-PE (Southern Biotech), or Goat F(ab′)2 anti-Mouse IgG2a, Human ads-PE (Southern Biotech) by flow cytometry (BD LSRFortessa X-20, Becton, Dickinson and Company). Gating strategy to determine the MFI is shown in Supplementary Fig. 11. MFI was calculated using BD FACSDiva Ver.8.0.1 software (Becton, Dickinson and Company). Staining antibodies used in flow cytometry experiments with experimental dilutions are listed in Supplementary Table 8.

Okura Y., Ikawa-Teranishi Y., Mizoroki A., Takahashi N., Tsushima T., Irie M., Harfuddin Z., Miura-Okuda M., Ito S., Nakamura G., Takesue H., Ozono Y., Nishihara M., Yamada K., Gan S.W., Hayasaka A., Ishii S., Wakabayashi T., Muraoka M., Nagaya N., Hino H., Nemoto T., Kuramochi T., Torizawa T., Shimada H., Kitazawa T., Okazaki M., Nezu J., Sollid L.M, & Igawa T. (2023). Characterizations of a neutralizing antibody broadly reactive to multiple gluten peptide:HLA-DQ2.5 complexes in the context of celiac disease. Nature Communications, 14, 8502.

+ Open protocol

+ Expand

Expansion of MAIT Cells in Vitro

Check if the same lab product or an alternative is used in the 5 most similar protocols

Vα7.2⁺ cells were stained with 1.25 μM CTV (Thermo Fisher Scientific Life Sciences), according to the manufacturer’s instructions. CTV-labeled Vα7.2⁺ cells were then cultured at 2 × 10⁵ cells/well for 3, 5, or 7 d in complete medium with monocytes (Vα7.2⁺ cell:monocyte ratio of 2:1) and fixed E. coli at the indicated microbial dose and in the presence of 1.25 μg/ml anti-CD28 mAb and 20 μg/ml anti-MR1 mAb (26.5; BioLegend) or IgG2a isotype control (ctrl) (MOPC-173; BioLegend). After 24 h, recombinant human IL-2 (PeproTech, Rocky Hill, NJ, USA) was added at a final concentration of 100 IU/ml and replenished every 48 h. Anti-MR1 mAb and IgG2a isotype ctrl were replenished at half the original concentration 72 h after they were first added and 48 h later. An expansion index of 0 was attributed to MAIT cells cultured without E. coli or for 3 d in the presence of anti-MR1 and at any microbial dose, because of the lack of discernible CTV dilution peaks in these conditions. All other expansion indices were determined by the FlowJo software v. 9.8 (Tree Star, Ashland, OR, USA).

Dias J., Sobkowiak M.J., Sandberg J.K, & Leeansyah E. (2016). Human MAIT-cell responses to Escherichia coli: activation, cytokine production, proliferation, and cytotoxicity. Journal of Leukocyte Biology, 100(1), 233-240.

+ Open protocol

+ Expand

Bifidobacteria modulate cytokine expression and ETEC response in PIE cells

Check if the same lab product or an alternative is used in the 5 most similar protocols

PIE cells were seeded at 3 × 10⁴ cells/12-well plate on type I collagen-coated
plates (Iwaki, Tokyo, Japan) and cultured for 3 days. After changing the medium,
bifidobacteria (5 × 10⁷ cells/ml) were added for 48 hr, and the cytokine
expression was analyzed. Similarly, in an ETEC challenge experiment, bifidobacteria (5 ×
10⁷ cells/ml) were added, and 48 hr later, each well was washed vigorously
with medium at least three times to eliminate all stimulants; and then cells were
stimulated with heat-stable ETEC PAMPs (equivalent to 5 × 10⁷ cells/ml) for 12
hours. In a blocking experiment, unlabeled anti-human TLR2-mouse IgG (Clone #TL2.1, Cat.
#309710, Biolegend, San Diego, CA, USA) and its isotype control antibody (MOPC-173, Cat.
#400224) were used. Cultured PIE cells were incubated with the unlabeled anti-TLR2 or
isotype control antibody for 12 hr before stimulation with bacteria.

MURATA K., TOMOSADA Y., VILLENA J., CHIBA E., SHIMAZU T., ASO H., IWABUCHI N., XIAO J.Z., SAITO T, & KITAZAWA H. (2014). Bifidobacterium breve MCC-117 Induces Tolerance in Porcine Intestinal Epithelial Cells: Study of the Mechanisms Involved in the Immunoregulatory Effect. Bioscience of Microbiota, Food and Health, 33(1), 1-10.

+ Open protocol

+ Expand

About PubCompare

Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.

We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.

However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.

Ready to get started?

Revolutionizing how scientists
search and build protocols!