Anti mouse pd 1 clone rmp1 14

Manufactured by BioXCell

Sourced in United States

Anti-mouse PD-1 (clone RMP1–14) is a laboratory-grade antibody used in research applications. It recognizes the mouse PD-1 (Programmed Cell Death 1) protein, which is involved in the regulation of the immune system. This antibody can be used in various immunological assays and experiments, but its specific applications should be determined by the user based on their research objectives.

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Spelling variants of this product

RMP1-14 (124 citations) Anti-PD-1 (65 citations) Anti-PD-1 (clone RMP1-14, (54 citations) Clone RMP1-14 (36 citations) Anti-mouse PD-1 (13 citations) Anti-mouse PD-1 antibody (clone RMP1-14) (6 citations) Rat anti-mouse PD-1 (clone RMP1–14; (3 citations) Anti-mouse PD-1 (CD279) (clone RMP1-14) (3 citations) Anti-mouse PD-1 Ab (anti-PD-1 Ab; clone RMP1- 14) (2 citations) InVivoPlus anti-mouse PD-1 (clone RMP1-14; (2 citations)

15 protocols using anti mouse pd 1 clone rmp1 14

Vaccine Delivery and Efficacy Testing

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Vaccines were administered via intramuscular injections in the quadriceps. A volume of 100 μL containing 5×10⁸ vp was delivered (50 μL per side). For efficacy studies, anti-mouse PD1 (clone RMP114, BioXCell) was administered intraperitoneally, at a dosage of 200 μg twice a week, starting from day 0 until day 16.

Garzia I., Nocchi L., Avalle L., Troise F., Leoni G., Seclì L., Antonucci L., Cotugno G., Allocca S., Romano G., Conti L., Caiazza C., Mallardo M., Poli V., Scarselli E, & D'Alise A.M. (2024). Tumor Burden Dictates the Neoantigen Features Required to Generate an Effective Cancer Vaccine. Cancer Immunology Research, 12(4), 440-452.

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Tumor Inoculation and Immunization in Mice

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Female C57BL/6 mice aged 8–12 weeks were acquired from The Jackson Laboratory. Tumor inoculation was performed by subcutaneously (s.c.) injecting animals with either 5 × 10⁵ E.G7-OVA, 10⁵ B16-F10, or 10⁵ MC-38 cells in the right flank. Immunizations were administered at a dose of either 6 nmol (OVA1/2 & Adpgk I/II) or 9 nmol (M27/30) of each antigen and CpG by s.c. injection in the abdomen. Immunizations were administered as listed in the treatment schedule provided in respective figures by s.c. injection in the abdomen. For combination therapy with the immune checkpoint inhibitor anti-PD-1, mice were administered 100 μg anti-mouse PD-1 (clone RMP1–14, BioXCell) via intraperitoneal injection. Tumor growth was measured on pre-determined days and volume was calculated using the following equation: tumor volume = length × width² × 0.5. Animals were euthanized when tumor volumes reached either 2,000 mm³ (E.G7-OVA) or 1,500 mm³ (B16-F10, MC-38) or when the animal became moribund.

Xie S., Green J., Bixby J.B., Szafranska B., DeMartini J.C., Hecht S, & Roberts R.M. (1997). The diversity and evolutionary relationships of the pregnancy-associated glycoproteins, an aspartic proteinase subfamily consisting of many trophoblast-expressed genes. Proceedings of the National Academy of Sciences of the United States of America, 94(24), 12809-12816.

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Entinostat and NHS-murine IL12 Combination Therapy

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Entinostat and recombinant NHS-murine IL12 were provided by Syndax and EMD Serono, respectively, under Cooperative Research and Development Agreements with the National Cancer Institute (NCI). Low-fat diet of 35% sucrose was enriched with entinostat for a target dose of 6 mg/kg/day (Research Diets). Anti-mouse PD-1 (clone RMP1-14, no. BE0146), PD-L1 (clone 10F.9G2, no. BE0101), rat IgG2a isotype (clone 2A3, no. BE0089), CD8α (clone 2.43, no. BE0061), and CD4 (clone GK1.5, no. BP0003-1) antibodies were purchased from BioXCell. Rabbit anti-asialo GM1 was purchased from Wako Chemicals USA.

Minnar C.M., Chariou P.L., Horn L.A., Hicks K.C., Palena C., Schlom J, & Gameiro S.R. (2022). Tumor-targeted interleukin-12 synergizes with entinostat to overcome PD-1/PD-L1 blockade-resistant tumors harboring MHC-I and APM deficiencies. Journal for Immunotherapy of Cancer, 10(6), e004561.

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Immunohistochemistry and Western Blot Analysis

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VPA was purchased from Enzo Life Sciences. Stock solutions were prepared in sterile water. Entinostat (MS-275) and vorinostat were from Selleck Chemicals; panobinostat (LBH589) from Novartis International; 5-Azacytidine (azacitidine) was from Sigma; GSK126 was from Active Biochem. Stock solutions were prepared in DMSO. Monoclonal antibodies anti-mouse PD-1 (clone RMP1-14, #BE0146) and anti-mouse CTLA-4 (clone 9H10, #BE0131) were purchased from Bioxcell. Actinomicin D was purchased from Sigma Aldrich.
All media, serum, antibiotics, and glutamine were from Corning.
Primary antibodies (Abs) for western blotting: β-Actin-Ab (Sigma-Aldrich, cod.A5316), Programmed death-ligand 1 (PD-L1)-Ab (Abcam, cod.Ab58810); (PD-L1)-Ab (cod.#13684), acetyl-H3-Ab (cod.#9649), PARP-Ab (cod.#9542) (Cell signaling Technology), and acetyl-H4-Ab (Millipore cod.06946). For IHC: monoclonal anti-mouse Ki67-Ab (Cell signaling Technology; cod.#12202), monoclonal anti-mouse CD4-Ab (Abcam, cod.Ab183685), anti-mouse CD8-Ab (eBioscience, clone 56-6.7). For immunofluorescence on fresh frozen tissues: anti-Foxp3-efluor570 (eBioscience clone FJK-16s, #41-5773-80).

Terranova-Barberio M., Thomas S., Ali N., Pawlowska N., Park J., Krings G., Rosenblum M.D., Budillon A, & Munster P.N. (2017). HDAC inhibition potentiates immunotherapy in triple negative breast cancer. Oncotarget, 8(69), 114156-114172.

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Combination Immunotherapy for Murine Colon Cancer

Cited 1 time

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Mice were housed and maintained by the Center for Comparative Medicine following animal protocols approved by the Massachusetts General Hospital Institutional Animal Care and Use Committee. CT26 murine colon carcinoma cells were acquired from ATCC (Manassas, VA) and cultured in RPMI. Monthly mycoplasma testing was performed by PCR screening and cells were discarded after 15 passages. All cell-based experiments were done with cells acquired within 6 months from ATCC during 2016 in order to ensure fidelity of the cell line identity. 1 × 10⁶ CT26 cells were prepared in a 1:1 (v:v) ratio in Matrigel (Corning, Tewksbury, MA) and injected subcutaneously into the right shoulder of Balb/C mice. Anti-mouse PD-1 (clone RMP1–14) and anti-mouse CTLA-4 (clone 9D9) were obtained from Bio X Cell (West Lebanon, NH). All mice were treated by intraperitoneal injection of saline (vehicle), 200 μg anti-PD1 (monotherapy), or both 200 μg anti-PD1 and 100 μg anti-CTLA-4 (combination therapy) on days 3, 6, and 9 following CT26 tumor inoculation(17 (link)).

Larimer B.M., Wehrenberg-Klee E., Dubois F., Mehta A., Kalomeris T., Flaherty K., Boland G, & Mahmood U. (2017). Granzyme B PET Imaging as a Predictive Biomarker of Immunotherapy Response. Cancer research, 77(9), 2318-2327.

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Characterization of 4T1 Mouse Mammary Tumor Cells

Cited 2 times

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Mouse mammary tumor cell line, 4T1, was purchased from ATCC (Manassas, VA), and maintained in the lab as described previously [21 (link), 6 (link), 12 (link)]. This cell line was retested recently and confirmed to be free of Ectromelia, EDIM, LCMV, LDEV, MHV, MNV, MPV, MVM, Mycoplasma pulmonis, Mycoplasma spp., Polyoma, PVM, REO3, Sendai, and TMEV by IMPACT III PCR Profile (IDEXX BioAnalytics, Columbia, MO). Engineering of oncolytic adenoviruses in this study (Ad(E-).null, Ad.sT, AdLyp.sT, mHAd.sT, and mHAdLyp.sT) was described in our previous publications [22 (link)–24 (link), 6 (link)]. Good Laboratory Practice (GLP) mass productions of them for animal studies were prepared by Gene Vector Core of Baylor College of Medicine (Houston, TX). Anti- mouse PD-1 (clone RMP1-14) and anti- mouse CTLA-4 (clone 9H10) antibodies were purchased from Bio X Cell (Lebanon, NH). All other materials used in this study were purchased from the vendors based on technique requirement and their past performance in the laboratory [6 (link), 12 (link), 16 (link), 25 (link)–28 (link)].

Xu W., Shin S.C., Vickman R., Filimon B., Yang Y., Hu Z., Mangold K., Prabhakar B, & Schreiber H. (2023). The Safety and Efficacy of Systemic Delivery of a New Liver-de-targeted TGFβ Signaling Inhibiting Adenovirus in an Immunocompetent Triple Negative Mouse Mammary Tumor Model. Research Square.

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Combination Immunotherapy for Melanoma

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B16F10-OVA cells (0:1 × 10⁶) were implanted intradermally on day 0. After 7 days mice were treated with 1 million OT-I CD8+ T-cells (sorted on chromatography channel or unsorted) i.v. On day 8 and 11, mice were i.t injected with 150 μ g of anti-mouse PD-1 (clone RMP1–14; BioXCell) in 30 μ l of saline.

Camargo C.P., Muhuri A.K., Alapan Y., Sestito L.F., Khosla M., Manspeaker M.P., Smith A.S., Paulos C.M, & Thomas S.N. (2023). Adhesion analysis via a tumor vasculature-like microfluidic device identifies CD8+ T cells with enhanced tumor homing to improve cell therapy. Cell reports, 42(3), 112175.

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Combination Immunotherapy for Melanoma

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Vesper S.J., Magnuson M.L., Dearborn D.G., Yike I, & Haugland R.A. (2001). Initial Characterization of the Hemolysin Stachylysin from Stachybotrys chartarum. Infection and Immunity, 69(2), 912-916.

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Liposomal Echinomycin and Immune Checkpoint Inhibitors

Cited 1 time

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Echinomycin was formulated with liposomes as previously described (35 (link)). Recombinant ipilimumab was provided by Lakepharma Inc. Remaining therapeutic antibodies were from BioXCell as follows: anti–mouse CTLA-4, clone 9D9 (BE0164); anti–mouse PD-1, clone RMP1-14 (BE0146); anti–mouse IFN-γ, clone XMG1.2 (BE0055); anti–mouse CD4, clone GK1.5 (BE0003-1); anti–mouse CD8α, clone YTS 169.4 (BE0117); and anti–mouse NK1.1, clone PK136 (BE0036).

Bailey C.M., Liu Y., Liu M., Du X., Devenport M., Zheng P., Liu Y, & Wang Y. (2022). Targeting HIF-1α abrogates PD-L1–mediated immune evasion in tumor microenvironment but promotes tolerance in normal tissues. The Journal of Clinical Investigation, 132(9), e150846.

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Antibody Targeting of PD-1/PD-L1 Axis

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PS-targeting antibody chimeric mch1N11 is a mouse IgG2a-kappa with human variable heavy and light chain regions; C44 is an IgG2a isotype. Purified anti-mouse PD-1 (clone RMP1-14) was obtained from BioXcell (West Lebanon, NH, USA). All in-vivo antibodies tested negative for endotoxin with a limit of detection of 0.5 EU/ml of stock antibody. Antibodies for flow cytometer analysis were CD45 (clone 30-F11), CD3e (clone 17A2), CD4 (clone GK1.5), CD8a (clone 53–6.7), and CD25 (clone PC61.5), all obtained from Bioscience (San Diego, CA, USA). Antibodies for immunohistochemistry were anti-PD-L1 antibody (clone MIH6, catalog number ab80276; Abcam, Cambridge, MA, USA) and anti-PD-1 antibody (clone RMP1-14, catalog number ab63477; Abcam).

Gray M.J., Gong J., Hatch M.M., Nguyen V., Hughes C.C., Hutchins J.T, & Freimark B.D. (2016). Phosphatidylserine-targeting antibodies augment the anti-tumorigenic activity of anti-PD-1 therapy by enhancing immune activation and downregulating pro-oncogenic factors induced by T-cell checkpoint inhibition in murine triple-negative breast cancers. Breast Cancer Research : BCR, 18, 50.

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