Anti pd 1 antibody

Manufactured by BioLegend

Sourced in United States

The Anti-PD-1 antibody is a lab equipment product that functions as a receptor antagonist for the programmed cell death 1 (PD-1) protein. It is used for research purposes to study the PD-1 pathway and its role in immune responses.

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Lab products found in correlation

Isotype control, by BioLegend (1 mentions) Recombinant mouse ifn γ, by R&D Systems (1 mentions) Isotype control antibody, by BioLegend (1 mentions) Penicillin streptomycin, by Merck Group (1 mentions) Romidepsin, by Selleck Chemicals (1 mentions) Pd l1 neutralizing antibody, by BioLegend (1 mentions) Anti ctla 4, by BD (1 mentions) Eh12.2h7, by BioLegend (1 mentions) Anti human pdl1 antibody, by BD (1 mentions) Annexin 5 fitc, by Roche (1 mentions)

6 protocols using anti pd 1 antibody

Modulation of Astrocyte-Microglia Crosstalk

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For co-culture experiments, astrocytes and microglia were activated with 20 ng/ml recombinant mouse IFN-γ (R&D, #485-MI-100/CF) overnight. The next day, microglia were detached using TrypLE and seeded onto primary mouse astrocytes with 1 µg/ml anti-PD-1 antibody (Biolegend, #135202) or isotype controls (Biolegend, #402301). After 24 h, cells were detached using TrypLE and intracellular flow cytometry was performed. In case of supernatant experiments, astrocytes were activated with 20 ng/ml mouse IFN-γ in combination with or without 5 µM BMS202 for 24 h. After 24 h, medium was aspirated and fresh medium was added for another 24 h. After an additional 24 h, astrocyte-conditioned medium (ACM) was collected, cleared from debris by centrifugation and added onto activated microglia. After 24 h incubation, microglia were harvested for RNA isolation and RT-qPCR analysis.

Linnerbauer M., Beyer T., Nirschl L., Farrenkopf D., Lößlein L., Vandrey O., Peter A., Tsaktanis T., Kebir H., Laplaud D., Oellinger R., Engleitner T., Alvarez J.I., Rad R., Korn T., Hemmer B., Quintana F.J, & Rothhammer V. (2023). PD-L1 positive astrocytes attenuate inflammatory functions of PD-1 positive microglia in models of autoimmune neuroinflammation. Nature Communications, 14, 5555.

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Modulating PD1 Pathway in PBMC

Cited 1 time

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Freshly isolated PBMCs were cultured in 24-well plates at 2 × 10⁶ cells/well in RPMI 1640 containing 1% penicillin/streptomycin (Sigma), L-Glutamine and 10% heat-inactivated human serum, with or without γ-Mtb CFP-10 + ESAT-6 (10 μg/ml) at 37 °C in a humidified 5% CO₂ atmosphere. To neutralize PD1, 10 μg/ml anti-PD1 antibody (BioLegend) was added to the cells in presence of the antigen. Isotype control antibody μg/ml (BioLegend) was added to some cells this served as a control well for PD1 specific inhibition. After 96 h, cell-free culture supernatants were collected, aliquoted and stored at − 70 °C until cytokine concentrations were measured by ELISA as published in [8 (link)]. Cells were washed and stained for surface and intracellular markers.

Devalraju K.P., Neela V.S., Ramaseri S.S., Chaudhury A., Van A., Krovvidi S.S., Vankayalapati R, & Valluri V.L. (2018). IL-17 and IL-22 production in HIV+ individuals with latent and active tuberculosis. BMC Infectious Diseases, 18, 321.

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Dual Tumor Regression via cGAMP-MPs and ICB

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2×10⁵ B16F10 cells were subcutaneously injected into the right rear flank of C57BL/6 female mice at day 0. Another 2×10⁵ B16F10 cells were subcutaneously injected into the left rear flank at day 2 to mimic metastatic tumor. Seven days after primary tumor inoculation, B16F10 tumor-bearing mice were divided into 4 experimental groups (n = 8 for each group): untreated, 1×cGAMP-MPs, 3×ICB (anti-PD-1), and 1×cGAMP-MPs+3×ICB. cGAMP-MPs (10 μg of cGAMP-S, 5 PLGA-1 particles containing a total of 10 μg of cGAMP, and 5 PLGA-2 particles containing a total of 10 μg of cGAMP) in 50 μL MC solution were intratumorally injected into the primary tumor (on the right side). For 3×ICB and 1×cGAMP-MPs+3×ICB treated groups, 100 μg of anti-PD-1 antibody (Biolegend, cat. no. 114114) was intraperitoneally injected at days 7, 11, and 15 after primary tumor inoculation. The distant tumor (left side) did not receive any treatments. Tumor size was measured every other day starting at day 7 after tumor inoculation with a digital caliper. Tumor volume was calculated using the following formula: length (mm) × width² (mm) × 0.5. Animals were euthanized when they showed signs of poor health or when the tumor size on either side exceeded 1500 mm³.

Lu X., Miao L., Gao W., Chen Z., McHugh K.J., Sun Y., Tochka Z., Tomasic S., Sadtler K., Hyacinthe A., Huang Y., Graf T., Hu Q., Sarmadi M., Langer R., Anderson D.G, & Jaklenec A. (2020). Engineered PLGA microparticles for long-term, pulsatile release of STING agonist for cancer immunotherapy. Science translational medicine, 12(556), eaaz6606.

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Mouse Colon Cancer Cell Lines and Romidepsin Treatment

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The mouse colon cancer cell lines CT26 and MC38 were obtained from the ATCC (Manassas, VA, USA) and cultured in RPMI-1640 with 10% FBS. All cells were maintained under humidified condition (37 °C, 5% CO₂), and continual culture did not exceed 2 months. Romidepsin was purchased from Selleck Chemicals (Houston, TX, USA). An anti-PD-1 antibody was purchased from Biolegend (San Diego, CA, USA). Nontarget and BRD4-targeting siRNAs were purchased from RiboBio Co., Ltd. (Guangdong, China).

Shi Y., Fu Y., Zhang X., Zhao G., Yao Y., Guo Y., Ma G., Bai S, & Li H. (2020). Romidepsin (FK228) regulates the expression of the immune checkpoint ligand PD-L1 and suppresses cellular immune functions in colon cancer. Cancer Immunology, Immunotherapy, 70(1), 61-73.

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Neutralizing PD-1/PD-L1 and CTLA-4 Suppression

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To investigate neutralizing the suppressor effect, purified anti-PD-1 antibody (0.5 and 1 μg/mL; Biolegend) was added in the same culture conditions described for the suppression assay above, using the corresponding isotype-matched mAb as control. Purified PD-L1 neutralizing antibody (Biolegend) was used at 5 μg/mL. To test involvement of CTLA-4 added together with anti-PD-1 in the suppression mechanism, 5 μg/mL of anti-CTLA-4 (BD Biosciences) was used.

Machicote A., Belén S., Baz P., Billordo L.A, & Fainboim L. (2018). Human CD8+HLA-DR+ Regulatory T Cells, Similarly to Classical CD4+Foxp3+ Cells, Suppress Immune Responses via PD-1/PD-L1 Axis. Frontiers in Immunology, 9, 2788.

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Tumor-Lymphocyte Apoptosis Regulation

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To examine the effect of tumor cells on lymphocyte apoptosis, 5×10⁶ MDA-MB-231 SCR cells and KD cells were cocultured with 5×10⁵ Jurkat leukemia T cells in the presence of mouse IgG (2 or 5 μg/ml; 17314; IBL Co., Ltd., Gunma, Japan), anti-human PD-L1 antibody (5 μg/ml; MIH1; BD) or anti-PD-1 antibody (2 μg/ml; EH12.2H7; BioLegend) in a 6-well plate for 24 hours. To block the PD-1/PD-L1 pathway, SCR cells were preincubated with anti-human PD-L1 antibody (5 μg/ml; BD) for 30 min or Jurkat cells were preincubated with anti-PD-1 antibody (2 μg/ml; BioLegend) for 30 min. Extent of apoptosis in Jurkat cells was determined by flow cytometry using FITC-Annexin V (11858777001; Roche) according to the manufacturer's instructions.

Tanaka T., Kutomi G., Kajiwara T., Kukita K., Kochin V., Kanaseki T., Tsukahara T., Hirohashi Y., Torigoe T., Okamoto Y., Hirata K., Sato N, & Tamura Y. (2017). Cancer-associated oxidoreductase ERO1-α promotes immune escape through up-regulation of PD-L1 in human breast cancer. Oncotarget, 8(15), 24706-24718.

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