Isotype control antibodies and blockade of Fc receptor Abs were provided from BioLegend (San Diego, CA, USA). Anti-CD3 (17A2), anti-CD11c (N418), anti-CD40 (HM40-3), anti-CD49b (DX5), anti-CD80 (16-10A1), anti-CD69 (H1.2F3), anti-CD86 (GL-1), anti-CD253 (TRAIL, N2B2), anti-granzyme B (GB11), anti-IFN-γ (R4-6A2), anti- IL-6 (MP5-20F3), anti-IL-12/23p40 (C17.8), anti- MHC class I (AF6-88.5), anti-MHC class II (M5/114.15.2), anti-NK1.1 (PK136), anti-perforin (S16009A), anti-TCR-β (H57-597) and anti-TNF-α (MP6-XT22) were purchased from BioLegend (San Diego, CA, USA). Anti- PD-L1 Abs were purchased from BioXcell (Lebanon, NH, USA). LPS (O111:B4) was purchased from Sigma-Aldrich.
Anti pd l1 ab
Manufactured by BioXCell
Sourced in United States
Anti-PD-L1 Ab is a monoclonal antibody that binds to the Programmed Death-Ligand 1 (PD-L1) protein. PD-L1 is a checkpoint protein expressed on the surface of certain cells that helps regulate the immune response.
Automatically generated - may contain errors
Lab products found in correlation
Lps o111 b4, by Merck Group (2 mentions)
Anti tnfα mp6 xt22, by BioLegend (1 mentions)
Anti tcrβ h57 597, by BioLegend (1 mentions)
Isotype control antibody, by BioLegend (1 mentions)
Isotype control, by BioLegend (1 mentions)
Cd3 fitc, by BioLegend (1 mentions)
Apc cyanine7 cd8, by BioLegend (1 mentions)
Fitc tnf α mp6 xt22, by BioLegend (1 mentions)
Percp cyanine5.5 cd11b, by BioLegend (1 mentions)
Pe cyanine7 cd86, by BioLegend (1 mentions)
Cd40 fitc, by BioLegend (1 mentions)
Percp cyanine5, by BioLegend (1 mentions)
Rat igg2b, by BioXCell (1 mentions)
Anti pd 1 ab, by BioXCell (1 mentions)
Rat igg2a, by BioXCell (1 mentions)
Female c57bl 6 mice, by Jackson ImmunoResearch (1 mentions)
Igg2a isotype control, by BioXCell (1 mentions)
Matrigel, by Corning (1 mentions)
C57bl 6j mice, by Japan SLC (1 mentions)
5 protocols using anti pd l1 ab
1
Immune Cell Activation Antibodies
2
Multicolor Flow Cytometry Panel
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LPS (O111:B4) was obtained from Sigma-Aldrich. Isotype controls and Fc receptor-blocking Abs were obtained from BioLegend (San Diego, CA, USA). FITC-CD3 (17A2, 1:40 dilution with PBS), PerCP5.5-CD4 (GK1.5, 1:50 dilution with PBS), APC/Cyanine7-CD8 (53-5.8, 1:40 dilution with PBS), PerCP/Cyanine5.5-CD11b (M1/70, 1:40 dilution with PBS), ACP/Cyanine7-CD11c (N418, 1:25 dilution with PBS), FITC-CD40 (HM40-3, 1:50 dilution with PBS), ACP/Cyanine7-CD69 (H1.2F3, 1:40 dilution with PBS), Brilliant Violet 605-CD80 (16-10A1, 1:25 dilution with PBS), PE/Cyanine7-CD86 (GL-1, 1:50 dilution with PBS), Alexa Fluor 647-CD197 (C-C chemokine receptor 7;CCR7, 4B12, 1:40 dilution with PBS) PerCP/Cyanine5.5-CD253 (tumor necrosis factor-related apoptosis-inducing ligand; TRAIL, N2B2, 1:25 dilution with PBS), Alexa Flour 647-granzyme B (GB11), PE/Cyanine7-IFN-γ (XMG1.2), PerCP/Cyanine5.5-major histocompatibility complex (MHC) class I (AF6-88.5, 1:40 dilution with PBS), PE-MHC class II (M5/114.15.2, 1:40 dilution with PBS), Brilliant Violet 510-NK1.1 (PK136, 1:40 dilution with PBS), PE-perforin (S16009A) and FITC-TNF-α (MP6-XT22) were also purchased from BioLegend. Anti-PD-L1 Abs were purchased from BioXcell (Lebanon, NH, USA). For the depletion assay of CD8 (YTS169.4) and NK1.1 (PK136)-positive cells, anti-CD8 and anti-NK.1 Abs were obtained from BioXcell.
3
Curcumin Enhances Anti-PD-1/PD-L1 Therapy
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For combination with PD‐1/PD‐L1 blockade, mice received po administration with 5 mg/kg of curcumin diluted in water every day starting on day 3 after implantation with MC38 until killed (day 20‐25). Control mice received water. Anti‐PD‐1 Ab (BioXcell, clone: RMP1‐14) and anti‐PD‐L1 Ab (BioXcell, clone: 10F.9G2) were given ip on days 7, 10, and 13 at a dose of 200 µg/100 µL/mouse. Rat IgG2a (BioXcell, clone: 2A3) and rat IgG2b (BioXcell, clone: LTF‐2) were injected ip in the control group and the curcumin single treatment group on days 7, 10, and 13 at a dose of 200 µg/100 µL/mouse.
4
Anti-PD-L1 Immunotherapy in Murine Tumors
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Animal experiments were approved by the Japanese Foundation for Cancer Research, Institutional Animal Care and Use Committee, and were conducted in accordance with the institutional guidelines. Details of the housing and husbandry of the mice have been previously described.
11 (link) Six‐week‐old C57BL/6 female mice (Jackson Laboratory) were subcutaneously injected with 100 μL of a cell solution prepared as 1 × 105 cells in a 1:1 mixture of Hanks’ balanced salt solution and Matrigel (Corning) of the same quantity. When tumors reached an average size of ~90 mm3, animals were randomized into six groups (10 animals/group/experiment in two independent experiments). Mice were treated with intraperitoneal injections of anti‐PD‐L1 Ab (Cat: #BE0101; BioXcell) or IgG2a isotype control (Cat: #BE0089; BioXcell) at 1.5 mg/kg or 5 mg/kg in phosphate‐buffered saline (PBS) twice a week for 2 weeks. The length (L) and width (W) of the tumor mass were measured and the TV was calculated using the following formula: TV = (L × W2)/2. Procedures for the treatment with clodronate liposomes (CL) were described in AppendixS1 . At the end of the experiments, mice were euthanized by cervical dislocation.
11 (link) Six‐week‐old C57BL/6 female mice (Jackson Laboratory) were subcutaneously injected with 100 μL of a cell solution prepared as 1 × 105 cells in a 1:1 mixture of Hanks’ balanced salt solution and Matrigel (Corning) of the same quantity. When tumors reached an average size of ~90 mm3, animals were randomized into six groups (10 animals/group/experiment in two independent experiments). Mice were treated with intraperitoneal injections of anti‐PD‐L1 Ab (Cat: #BE0101; BioXcell) or IgG2a isotype control (Cat: #BE0089; BioXcell) at 1.5 mg/kg or 5 mg/kg in phosphate‐buffered saline (PBS) twice a week for 2 weeks. The length (L) and width (W) of the tumor mass were measured and the TV was calculated using the following formula: TV = (L × W2)/2. Procedures for the treatment with clodronate liposomes (CL) were described in Appendix
5
Evaluating Combination Therapy for Liver Cancer
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Male wild-type C57BL/6J mice 8-weeks of age were obtained from Japan SLC (Shizuoka, Japan). After making a small incision under anesthesia to expose the spleen, 0.1 mL of a viable cell suspension containing 5 × 106 cells/mL was injected into the spleen. We chose SL4 cells because the cells grow rapidly, even in the liver of wild-type mice [21 (link)]. The animals were then each intraperitoneally injected with or without 0.4 mg PRI-724 (Prism Pharma, Yokohama, Japan) and/or 200 μg of an anti-PD-L1 Ab (10F.9G2; Bio X Cell, NH, USA) three times per week. In addition, some mice treated with PRI-724 and the anti-PD-L1 Ab were administrated anti-mouse CD4 or CD8 Ab (250 μg/mouse; Bio X Cell) three times per week. After the course of treatment, the mice were anesthetized and humanely sacrificed by exsanguination 14 days post cell-inoculation. The livers of the animals were immediately removed, washed in ice-cold PBS, and weighed before a portion of the dissected liver tissue was frozen in liquid nitrogen. Additional animals were maintained and used for survival analysis.
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