Mouse Lewis lung carcinoma cell line (LLC) was purchased from Shanghai Cell Bank of Chinese Academy of Sciences. The MPE mouse model (C57BL/6J, six-week-old, male; Model Animal Research Center of Nanjing University) has been previously described (18 (link), 20 (link)) ( Figure S1 ) . Mice were reportedly administered with an intrathoracic injection of 200μg anti-PD1 mAb (InVivoMab anti-mouse PD-1 (CD279), Bioxcell)/0.9% normal saline on the 7th and 14th day of the model. We then injected 50 μL anti-CD8 mAb (4 mg/kg, InVivoPlus anti-mouse CD8A, Bioxcell) intraperitoneally multiple times to achieve CD8 depletion. The fixed mouse tissues were successively scanned using a CT machine (Siemens Somatom Sensation 16).
Invivomab anti mouse pd 1 cd279
Manufactured by BioXCell
Sourced in United States
InVivoMab anti-mouse PD-1 (CD279) is a laboratory product designed for research purposes. It is an antibody that binds to the mouse PD-1 protein, which is a cell surface receptor involved in immune system regulation.
Automatically generated - may contain errors
Lab products found in correlation
Invivomab rat igg2a isotype control, by BioXCell (5 mentions)
Penicillin, by Thermo Fisher Scientific (2 mentions)
Streptomycin, by Thermo Fisher Scientific (2 mentions)
Somatom sensation 16, by Siemens (1 mentions)
Pembrolizumab, by Selleck Chemicals (1 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (1 mentions)
3 4 5 dimethylthiazol yl 2 5 diphenyltetrazolium bromide mtt, by Merck Group (1 mentions)
Trypsin, by Thermo Fisher Scientific (1 mentions)
Anti erk1 2, by Santa Cruz Biotechnology (1 mentions)
Antibody against gapdh, by Proteintech (1 mentions)
Anti pd l1, by Santa Cruz Biotechnology (1 mentions)
Anti phospho erk1 2, by Santa Cruz Biotechnology (1 mentions)
P erk1 2, by Santa Cruz Biotechnology (1 mentions)
Dulbecco modified eagle medium (dmem), by Thermo Fisher Scientific (1 mentions)
Pd0325901, by Selleck Chemicals (1 mentions)
Dimethyl sulfoxide (dmso), by Merck Group (1 mentions)
Takinib, by Cayman Chemical (1 mentions)
Anti mouse cd8α, by Cell Signaling Technology (1 mentions)
5z 7 oxozeaenol, by Enzo Life Sciences (1 mentions)
Flag tag (flag), by Cell Signaling Technology (1 mentions)
9 protocols using invivomab anti mouse pd 1 cd279
1
Murine Lewis Lung Carcinoma Imaging
2
Investigating Immune Checkpoint Inhibition in Cancer
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PD0325901 and PD-1 blocking antibody (pembrolizumab) were purchased from Selleck Chemicals. Anti-PD-L1, anti-phospho ERK1/2 and anti-ERK1/2 antibodies were obtained from Santa Cruz Biotechnology. The antibody against GAPDH was purchased from Proteintech. The anti-mouse CD3 and granzyme B antibodies for immunohistochemistry were purchased from Abcam. The anti-human PD-L1 and p-ERK1/2 for immunohistochemistry were obtained from Santa Cruz Biotechnology. The anti-human CD3 and CD8 antibodies for immunohistochemistry were purchased from Zsbio. Dimethyl sulfoxide and 3-(4,5-dimethylthiazol-yl)-2,5-diphenyltetrazolium bromide (MTT) were products of Sigma–Aldrich. DMEM and fetal bovine serum were products of Gibco. Penicillin, streptomycin and trypsin were obtained from Thermo Fisher Scientific. InVivoMab anti-mouse PD-1 (CD279) was purchased from BioXcell.
3
Establishment and Evaluation of Murine Tumor Models
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Cells (5.0 × 105 cells in 100 μL phosphate-buffered saline) were mixed with 100 μL Matrigel Matrix Basement Membrane High Concentration (Corning, Inc., Corning, NY). The cells were injected subcutaneously into 8-week-old male NOD.CB17-Prkdcscid/J or C57BL/6 mice. For the establishment of orthotopic liver tumor models, the cells (1.0 × 106 cells in 100 μL phosphate-buffered saline) were injected via the spleen into 8-week-old male C57BL/6 mice. At 30 days after injection, hepatic tumors were evaluated by hematoxylin and eosin staining and immunohistochemistry.
Purified anti-mouse PD1 monoclonal antibody, InVivoMAb anti-mouse PD-1 (CD279) (catalog no. BE0146, clone: RMP1-14), and control Ig, InVivoMAb rat IgG2a isotype control (catalog no. BE0089, clone: 2A3), were purchased from BioXCell (Lebanon, NH). The anti-PD1 antibody (100 μg/mouse) was intraperitoneally administered to MHCF1- or MHCF5-derived tumor-bearing mice every 3 days from day 6 to 18 (total 5 times), and tumor volumes were evaluated until 60 days and 25 days, respectively.
Purified anti-mouse PD1 monoclonal antibody, InVivoMAb anti-mouse PD-1 (CD279) (catalog no. BE0146, clone: RMP1-14), and control Ig, InVivoMAb rat IgG2a isotype control (catalog no. BE0089, clone: 2A3), were purchased from BioXCell (Lebanon, NH). The anti-PD1 antibody (100 μg/mouse) was intraperitoneally administered to MHCF1- or MHCF5-derived tumor-bearing mice every 3 days from day 6 to 18 (total 5 times), and tumor volumes were evaluated until 60 days and 25 days, respectively.
4
Melanoma Xenograft Immunotherapy in Mice
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For the in vivo treatment model, 1.5 × 106 YUMMER1.7 cells in 50 µL of PBS were subcutaneously inoculated in the right hind paw of 6-week-old C57BL-6 mice. Cells were harvested by trypsinization and resuspended in PBS (pH 7.4) before animals were injected.
During inoculation, mice were kept under inhalational anesthesia with isoflurane 2% in 2 L/min airflow.
Tumor-bearing mice were injected intraperitoneally (i.p) with PD1 antibody or isotype control antibodies (10 mg/kg) 3 times per week (days0-2-4-7-9-11) for 2 weeks at a size of 300 ± 50 mm³. Antibodies (in vivo MAb rat IgG2a isotype control and in vivo MAb anti-mouse PD-1 (CD279)) were purchased from BioXcell (Lebanon, NH, USA). After the treatments were interrupted, tumor regrowth was longitudinally monitored using an electronic caliper and calculated via the formula: 4/3 × π×(L/2) × (l/2)2. The growth delay of melanoma xenografts was calculated as the time taken (in days) to reach a volume of 1000 mm3.
During inoculation, mice were kept under inhalational anesthesia with isoflurane 2% in 2 L/min airflow.
Tumor-bearing mice were injected intraperitoneally (i.p) with PD1 antibody or isotype control antibodies (10 mg/kg) 3 times per week (days0-2-4-7-9-11) for 2 weeks at a size of 300 ± 50 mm³. Antibodies (in vivo MAb rat IgG2a isotype control and in vivo MAb anti-mouse PD-1 (CD279)) were purchased from BioXcell (Lebanon, NH, USA). After the treatments were interrupted, tumor regrowth was longitudinally monitored using an electronic caliper and calculated via the formula: 4/3 × π×(L/2) × (l/2)2. The growth delay of melanoma xenografts was calculated as the time taken (in days) to reach a volume of 1000 mm3.
5
Investigating TAK1 Signaling Pathway
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The commercial antibodies used were ESRP2, β‐actin, TAK1, phospho‐TAK1 (Ser439), GSDME (Abcam), p38, phospho‐p38, JNK, phospho‐JNK, p65, phospho‐p65, IKK, phospho‐IKK, cleaved caspase‐3, TAB1, TAB2, GSDMD, anti‐mouse CD8α, FLAG (Cell Signaling Technology), TAB3 (Santa Cruz Biotechnology), InVivoMAb anti‐mouse PD‐1 (CD279), and InVivoMAb rat IgG2a isotype control (Bio X Cell). The TAK1 inhibitor, takinib, was purchased from Cayman Chemical, and 5Z‐7‐oxozeaenol was obtained from Enzo Life Sciences.
6
Intraperitoneal Anti-PD1 Antibody Injection
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For intraperitoneal injections of anti-PD1 antibody, InVivoMAb anti-mouse PD1 (CD279) (BE0146, BioXCell) was diluted to 3 μg/μL in sterile PBS and 300 μg was intraperitoneally injected on day 37 and day 44 after locoregional APTKA implantation. IgG2a isotype control (BE0089, BioXCell) in the same dilution served as control.
7
Murine Immune Checkpoint Blockade Protocol
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The InVivoMab anti-mouse CD8alpha (BE0061), InVivoMab anti-mouse NK1.1 (BE0036), InVivoMab anti-mouse PD-1 (CD279) (BE0273), InVivoMab anti-mouse PD-L1 (B7-H1) (BE0101), InVivoMab rat IgG2a isotype control (BE0089), and InVivoMab rat IgG2b isotype control (BE0090) were purchased from Bio X Cell, diluted in InVivoPure pH 7.0 Dilution Buffer (IP0070), and administered as indicated in the corresponding figures. Rabbit anti-murine RANTES (CCL5) (500-P118) was purchased from PeproTech.
8
Anti-PD-1 Immunotherapy in Mice
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When tumors reached measurable size (ca. 30 mm3), mice were treated with 10 mg/kg of anti-PD-1 (InVivoMAb anti-mouse PD-1 (CD279), Bio X Cell) by intraperitoneal injection. Control animals were treated with 10 mg/kg of IgG2a isotype (InVivoMAb rat IgG2a isotype control, Bio X Cell). After the first injection, mice received injections every third day for a total of approximately 2–4 weeks, depending on the duration of the study.
9
HDAC6 Inhibitor Nexturastat A Protocol
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The HDAC6 selective inhibitor Nexturastat A (NextA) was purchased from StarWise Pharmaceuticals, Madison, Wisconsin, USA. NextA was kept at a stock solution of 10 mg/mL and diluted with a buffer provided by the manufacturer to the concentration used for each particular experiment. Tubastatin A (S8049) was purchased from Selleckchem. MS275 (50-148-306) was purchased from Biotang Inc. The pan-HDACi LBH589 (50-148-338) was purchased from Biotang Inc. In Vivo MAb anti-mouse PD-1 (CD279) (BE0146) was manufactured and purchased from Bio X Cell (West Lebanon, New Hampshire, USA) and stored at 4 °C for use throughout the study.
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