Pd l1 mih5

Manufactured by Thermo Fisher Scientific

Sourced in United States

The PD-L1 (MIH5) is a monoclonal antibody targeting the Programmed Death-Ligand 1 (PD-L1) protein. It is designed for use in research and experimental applications.

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Lab products found in correlation

H 2 db kh95, by BD (1 mentions) H 2kb af6 88.5, by BD (1 mentions) Hla abc w6 32, by BioLegend (1 mentions) Cd3 17a2, by Thermo Fisher Scientific (1 mentions) Cd4 gk1, by BioLegend (1 mentions) Cd62l mel 14, by Thermo Fisher Scientific (1 mentions) Cd44 im7, by BioLegend (1 mentions) Cd8a 53 6, by BioLegend (1 mentions) 7 aad, by Thermo Fisher Scientific (1 mentions) Lsrfortessa x 20, by BD (1 mentions) Facsverse flow cytometer, by BD (1 mentions) Cd8 53 6, by BD (1 mentions) Fcs express, by De Novo Software (1 mentions) Cd4 gk1, by BD (1 mentions) Cd1d 1b1, by BD (1 mentions) Cd5 53 7, by BD (1 mentions) Pd l2 ty25, by Thermo Fisher Scientific (1 mentions) Anti mouse cd19 1d3, by BD (1 mentions) Facscalibur flow cytometer, by BD (1 mentions) Fitc labeled annexin 5 propidium iodide apoptosis detection kit, by BD (1 mentions)

3 protocols using pd l1 mih5

Multiparametric flow cytometry for immune cells

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Samples were blocked with 2% normal mouse serum. Fixable yellow (L34959, Invitrogen, Carlsbad, CA, USA) or 7AAD (Cat 00699350, eBioscience, Waltham, MA, USA) was used to stain live/dead cells. Anti-mouse antibodies used were CD3 (17A2, eBioscience), CD4 (GK1.5, BioLegend, San Diego, CA, USA), TCR Vα2 (KB5-C20, BD Pharmingen), CD8a (53-6.7, BioLegend), CD44 (IM7, Biolegend), CD62L (MEL-14, eBioscience), H-2Db (KH95, BD Pharmingen), H-2Kb (AF6–88.5, BD Pharmingen), PD-L1 (MIH5, eBioscience). Anti-human antibodies used were HLA ABC (W6/32, Biolegend), PD-L1 (29E.283, Biolegend), anti-mouse IgG2a K Isotype (Biolegend), and anti-mouse IgG2b K (Biolegend).
Fluorescence was measured on BD LSR Fortessa X-20 or BD FACSVerse flow cytometer (BD Biosciences, North Ryde, NSW, Australia) and data analyzed using the FlowJo, LLC software (BD Biosciences, Franklin Lakes, NJ, USA).

Lau P.K., Cullinane C., Jackson S., Walker R., Smith L.K., Slater A., Kirby L., Patel R.P., von Scheidt B., Slaney C.Y., McArthur G.A, & Sheppard K.E. (2021). Enhancing Adoptive Cell Transfer with Combination BRAF-MEK and CDK4/6 Inhibitors in Melanoma. Cancers, 13(24), 6342.

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Phenotypic Characterization of Immune Cells

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The individual performing FACS analysis was not blinded to genotype. Anti-mouse Abs CD4 (GK1.5, BD Pharmingen, Franklin Lakes, NJ, USA) and CD8 (53-6.7, BD Pharmingen) were used for the proliferation assay. Anti-mouse CD19 (1D3, BD Pharmingen), CD1d (1B1, BD Pharmingen), CD5 (53-7.3, BD Pharmingen), CD28 (37.51, BD Pharmingen), CD152 (CTLA-4, UC10-4B9), ICOS (C398-4A, BD Pharmingen), PD-L1 (MIH5, eBioscience), and PD-L2 (TY25, eBioscience) were used for this study. Single-cell suspensions were washed with staining medium (PBS containing 0.1% NaN₃ and 2% FCS). After incubation with mAb and washing with staining buffer, propidium iodide (PI) was added to identify dead cells. FACS data acquisition was performed on a FACSCalibur flow cytometer (BD Biosciences, San Jose, CA, USA) and data were analyzed using isotype control Abs to set quadrants before calculating the percentage of positive cells, using FCS Express (De Novo Software, Los Angeles, CA, USA).

Bodhankar S., Chen Y., Lapato A., Vandenbark A.A., Murphy S.J, & Offner H. (2014). Targeting immune co-stimulatory effects of PD-L1 and PD-L2 might represent an effective therapeutic strategy in stroke. Frontiers in Cellular Neuroscience, 8, 228.

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Characterization of Immune Cell Phenotypes

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The cell surfaces was stained with antibodies at 4°C for 30 min. Specific antibodies against CD11b (M1/70), F4/80 (BM8), CD206 (MMR), PD-1 (J43) and PD-L1 (MIH5) were purchased from eBioscience (San Diego, CA). The samples were washed using PBS and fixed with 4% paraformaldehyde. The samples were analyzed using FACSCantoⅡ^TM flow cytometer (BD Biosciences, San Jose, CA). The data were analyzed using FlowJo software (Tree Star, Ashland, OR). Cell apoptosis was measured using a double staining with FITC-labeled Annexin V/Propidium Iodide (PI) Apoptosis Detection Kit (BD Biosciences) according to the manufacturer’s instruction. The apoptosis levels were examined by flow cytometry. For reactive oxygen species (ROS) measurement, the cells were pretreated with 10 mM NAC (Sigma-Aldrich, St. Louis, MO) for 24 h. Then, 10 μM DCF-DA (Sigma-Aldrich) was added to the cells for 30 min at 37°C, and DCF fluorescence was measured by flow cytometry.

Lee S., Lee A., Lim J, & Lim J.S. (2022). Regulation of tumor-associated macrophage (TAM) differentiation by NDRG2 expression in breast cancer cells. BMB Reports, 55(2), 81-86.

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