Anti-Mouse PD-1 [Clone RMP1-30] — Purified in vivo GOLD™ Functional Grade

Anti-Mouse PD-1 [Clone RMP1-30] — Purified in vivo GOLD™ Functional Grade

Product No.: C3442

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Clone
RMP1-30
Target
PD-1
Formats AvailableView All
Product Type
Monoclonal Antibody
Alternate Names
Programmed Death-1, CD279
Isotype
Rat IgG2b κ
Applications
Depletion
,
FC
,
in vivo

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Antibody Details

Product Details

Reactive Species
Mouse
Host Species
Rat
Recommended Isotype Controls
Recommended Dilution Buffer
Immunogen
Mouse PD-1 transfected BHK cells
Product Concentration
≥ 5.0 mg/ml
Endotoxin Level
<1.0 EU/µg as determined by the LAL method
Purity
≥95% monomer by analytical SEC
>95% by SDS Page
Formulation
This monoclonal antibody is aseptically packaged and formulated in 0.01 M phosphate buffered saline (150 mM NaCl) PBS pH 7.2 - 7.4 with no carrier protein, potassium, calcium or preservatives added. Due to inherent biochemical properties of antibodies, certain products may be prone to precipitation over time. Precipitation may be removed by aseptic centrifugation and/or filtration.
Product Preparation
Functional grade preclinical antibodies are manufactured in an animal free facility using in vitro cell culture techniques and are purified by a multi-step process including the use of protein A or G to assure extremely low levels of endotoxins, leachable protein A or aggregates.
Storage and Handling
Functional grade preclinical antibodies may be stored sterile as received at 2-8°C for up to one month. For longer term storage, aseptically aliquot in working volumes without diluting and store at ≤ -70°C. Avoid Repeated Freeze Thaw Cycles.
Country of Origin
USA
Shipping
Next Day 2-8°C
Applications and Recommended Usage?
Quality Tested by Leinco
FC
IHC FF
Additional Applications Reported In Literature ?
FC
Depletion
Each investigator should determine their own optimal working dilution for specific applications. See directions on lot specific datasheets, as information may periodically change.

Description

Description

Specificity
RMP1-30 activity is directed against mouse PD-1 (CD279).
Background
PD-1, a member of the CD28/CTLA-4 subfamily of the Ig superfamily, is a transmembrane protein expressed on activated T cells, B cells, a subset of thymocytes, macrophages, dendritic cells, and some tumor cells1,2. PD-1 is also retained in the intracellular compartments of human and mouse regulatory T cells (Tregs) and is co-expressed with CD25 on activated CD4+ T cells3. When stimulated via the T cell receptor (TCR), Tregs translocate PD-1 to the cell surface3. PD-1 is absent on naïve T cells and is inducibly expressed on T cells by T cell antigen receptor (TCR). B7-H1 (PD-L1; CD274) and B7-DC (PD-L2; CD273) have been identified as PD-1 ligands1. PD-1 is co-expressed with PD-L1 on tumor cells and tumor-infiltrating antigen-presenting cells (APCs)2.

PD-1 acts as a T cell inhibitory receptor and plays a critical role in peripheral tolerance induction and autoimmune disease prevention as well as important roles in the survival of dendritic cells, macrophage phagocytosis, and tumor cell glycolysis2. PD-1 prevents uncontrolled T cell activity, leading to attenuation of T cell proliferation, cytokine production, and cytolytic activities. Additionally, the PD-1 pathway, consisting of PD-1 on T cells and PD-L1 on APCs, is a major mechanism of tumor immune evasion, and, as such, PD-1 is a target of cancer immunotherapy2.

RMP1-30 does not block the binding of B7-H1 or B7-DC to PD-11. However, recent studies show that RMP1-30 does deplete PD-1 expressing cells 28.
Antigen Distribution
PD-1 is expressed on activated T cells, B cells, a subset of thymocytes, macrophages, dendritic cells, and some tumor cells.
NCBI Gene Bank ID
Research Area
Immunology

Leinco Antibody Advisor

Powered by AI: AI is experimental and still learning how to provide the best assistance. It may occasionally generate incorrect or incomplete responses. Please do not rely solely on its recommendations when making purchasing decisions or designing experiments.

Clone RMP1-30 is a rat monoclonal antibody specific for mouse PD-1 (CD279), and its common in vivo applications in mice are primarily for detection and characterization of PD-1 expression, such as via flow cytometry and immunophenotyping, rather than for functional blocking of PD-1 signaling.

Key in vivo uses of RMP1-30 in mice include:

  • Flow cytometric analysis of PD-1 expression on activated and unactivated T cells, B cells, dendritic cells, macrophages, and certain tumor cells.
  • Immunophenotyping of immune cell populations in mice to monitor PD-1 surface protein, especially in studies on immune regulation and tumor immunology.
  • Identification and isolation of PD-1+ cell populations (e.g., by FACS—fluorescence-activated cell sorting) for downstream analyses, such as transcript enrichment and functional studies.

Limitations and Important Notes:

  • RMP1-30 is not considered suitable for in vivo functional PD-1 blockade (i.e., it does not block the PD-1/PD-L1 interaction in live animals). For functional/therapeutic in vivo blockade, other clones like 29F.1A12 are used preferentially.
  • While RMP1-30 can bind PD-1 in in vivo settings, its primary role is as a detection and characterization tool rather than for manipulating PD-1-mediated immune pathways in therapeutic or mechanistic studies.

Summary Table: RMP1-30 in Vivo Applications in Mice

ApplicationDescriptionFunctional Blockade?
Flow cytometry & immunophenotypingLabels and monitors PD-1 expression on immune cell subsets and tumor cells in vivoNo
FACS sorting for downstream analysisIsolates PD-1+ cell populations from in vivo samples for transcriptomic or proteomic workNo
In vivo detection reagentUsed for the detection of PD-1 presence in various murine disease modelsNo

When planning experiments requiring functional modulation (blockade or agonism) of mouse PD-1 signaling in vivo, alternative clones should be selected.

RMP1-30 is commonly used in combination with several other antibodies and proteins in murine immune checkpoint research, particularly for validation, epitope mapping, and functional studies.

Anti-PD-1 Antibodies

29F.1A12 is one of the most frequently paired antibodies with RMP1-30. These two clones recognize overlapping populations, with co-staining studies showing that approximately 90-97% of PD-1-positive cells are dual positive for both antibodies. The 29F.1A12 antibody is particularly notable because it functions as a strong blocking antibody that can prevent PD-1/PD-L1 interactions, whereas RMP1-30 cannot block this interaction. This complementary functionality makes them useful together in experimental designs where one antibody blocks immune checkpoint signaling while the other serves as a detection reagent.

RMP1-14 is another anti-PD-1 antibody frequently used alongside RMP1-30. Like 29F.1A12, RMP1-14 has blocking capability, though it requires approximately 100-fold higher concentrations to achieve similar effects. RMP1-30 can be used to stain for PD-1 in experiments where RMP1-14 is employed therapeutically, though with an approximately 25% decrease in fluorescence when the therapeutic antibody is already bound to PD-1.

PD-L1 and Checkpoint Proteins

PD-L1 (also known as B7-H1) is frequently investigated in the same experimental systems as RMP1-30. As the primary ligand for PD-1, PD-L1 antibodies such as clone 10F.9G2 are often used in combination studies to understand the complete checkpoint interaction pathway. Additionally, PD-L2, the second ligand for PD-1, is relevant in these studies as it can also mediate inhibitory signals through PD-1 binding.

These antibody combinations are critical for dissecting immune checkpoint roles in health and disease, with researchers often using RMP1-30 for detection purposes while employing blocking antibodies like 29F.1A12 to modulate immune responses therapeutically.

Clone RMP1-30 is a monoclonal antibody against mouse PD-1 that has been widely cited in the scientific literature primarily as a PD-1 detection reagent in flow cytometry and immunophenotyping applications, but it does not function as a blocking antibody for the PD-1/PD-L1 interaction.

Key findings from scientific literature citations of RMP1-30:

  • Specificity: RMP1-30 binds specifically to PD-1 on the surface of murine immune cells and tumor cells, with high specificity confirmed by lack of binding to PD-1 knockout cells.
  • Epitope recognition and overlap: RMP1-30 and other anti-PD-1 clones (such as 29F.1A12 and RMP1-14) recognize overlapping but non-identical PD-1 epitopes. RMP1-30 can be used alongside 29F.1A12 and RMP1-14 for co-staining, with evidence for dual positivity in almost all PD-1^+^ cells.
  • Blocking function: RMP1-30 does not block the interaction between PD-1 and PD-L1, and does not reverse PD-1 mediated inhibition of T cell signaling. This contrasts with clones 29F.1A12 and RMP1-14, which are capable of blocking activity. Studies confirmed that RMP1-30 does not enhance T cell activation via PD-1 pathway blockade.
  • Use in combination with blocking antibodies: Because it is non-blocking, RMP1-30 is often used to stain or track PD-1 in the presence of blocking antibodies (e.g., 29F.1A12 or RMP1-14) to verify PD-1 status without interfering with the action of therapeutic or functional antibodies.
  • Application in tumor and immune cell phenotyping: RMP1-30 is used to detect PD-1 on live and dead tumor cells, T cells, and other immune cell types in both 2D and 3D culture systems, revealing context-dependent differences in expression.
  • Cooperative binding: There is some evidence of cooperative binding between RMP1-30 and certain other anti-PD-1 antibodies, potentially leading to increased fluorescence signal in antibody panels.

Additional notes:

  • Agonist activity: Recent studies suggest potential for RMP1-30 (or closely related antibodies) to exhibit PD-1 agonist activity in specific in vitro or in vivo contexts, possibly modulating inflammation. However, evidence for robust agonist function in vivo remains limited and may depend on experimental conditions.
  • Widespread citation: RMP1-30 is extensively cited as a key reagent in immuno-oncology studies, immune checkpoint validation, and comparative studies of anti–PD-1 antibodies in murine preclinical research.

In summary, RMP1-30 is a reliable, non-blocking anti–PD-1 clone for detection and phenotyping of murine PD-1, frequently used in combination antibody panels to dissect immune cell and tumor PD-1 expression, but is not used for functional PD-1 blockade in immunotherapy research.

Dosing regimens for clone RMP1-30 (anti-mouse PD-1 antibody) vary substantially across mouse models and research applications, with no single standardized protocol universally adopted. Most commonly, RMP1-30 is used for flow cytometry and immunophenotyping, not in vivo functional blockade; dosing and scheduling are therefore tied to experimental goals, cell numbers, and application type.

Key context for RMP1-30 dosing:

  • Primary applications: Unlike clones such as 29F.1A12 or RMP1-14, RMP1-30 is typically utilized as a non-blocking antibody to detect PD-1 expression (e.g., by flow cytometry), rather than functional blockade or therapeutic intervention.
  • Blocking activity: RMP1-30 does not block PD-L1 binding or stimulate T cell responses in vivo, so its dosing is usually not optimized for checkpoint blockade studies.

Dosing Regimen Variation

For flow cytometry:

  • Typical working dilutions range from 1:100 to 1:200 of antibody stock (usually at 1 mg/ml).
  • Commonly, 10 μl of dilution is used to stain 1 million cells in 100 μl buffer.

For in vivo use:

  • Dosing guidelines are rarely published for RMP1-30, and when used, regimens are tailored to model demands (e.g., mouse strain, research question, marker expression).
  • General recommendations for in vivo antibodies (other anti-PD-1 clones) are between 100–500 μg per mouse by intraperitoneal injection every 3–4 days, but these are specific to functional blocking clones and not directly applicable to RMP1-30.
  • Leinco notes that RMP1-30 dosing "is not standardized and can vary considerably across different mouse models depending on the research application" and should be "optimized by the end user".

Essential Points

  • No consensus dosing regimen exists for RMP1-30 in vivo; protocols are customized per experiment and are typically provided as starting points or require optimization by users.
  • Published studies using RMP1-30 may report diverse protocols based on model type (i.e., tumor, autoimmune, or infectious disease), but generally do not specify standardized doses as they would for blocking antibodies.
  • For detection or immunophenotyping, use recommended flow cytometry dilutions; for any functional or depletion study (rare for RMP1-30), start with comparable doses to other anti-PD-1 antibodies, but verify antibody activity and intent.

If your application is functional PD-1 blockade (e.g., immunotherapy or checkpoint studies), RMP1-30 is not appropriate due to its lack of blocking activity—use clones such as 29F.1A12 or RMP1-14 for those protocols.

In summary, doses of RMP1-30 are not standardized and vary with the mouse model and experimental objective, most commonly employed for PD-1 detection rather than in vivo modulation.

References & Citations

1. Matsumoto K, Inoue H, Nakano T, et al. J Immunol. 172(4):2530-2541. 2004.
2. Zhao Y, Harrison DL, Song Y, et al. Cell Rep. 24(2):379-390.e6. 2018.
3. Raimondi G, Shufesky WJ, Tokita D, et al. J Immunol. 176(5):2808-2816. 2006.
4. Ding ZC, Habtetsion T, Cao Y, et al. Sci Rep. 7(1):12168. 2017.
5. Chatterjee S, Daenthanasanmak A, Chakraborty P, et al. Cell Metab. 27(1):85-100.e8. 2018.
6. Snell LM, MacLeod BL, Law JC, et al. Immunity. 49(4):678-694.e5. 2018.
7. Bradley CP, Teng F, Felix KM, et al. Cell Host Microbe. 22(5):697-704.e4. 2017.
8. Uchil PD, Pi R, Haugh KA, et al. Cell Host Microbe. 25(1):87-100.e10. 2019.
9. Timilshina M, You Z, Lacher SM, et al. Cell Rep. 27(10):2948-2961.e7. 2019.
10. Chow MT, Ozga AJ, Servis RL, et al. Immunity. 50(6):1498-1512.e5. 2019.
11. St Clair JB, Detanico T, Aviszus K, et al. PLoS One.12(1):e0170556. 2017.
12. Liu QZ, Ma WT, Yang JB, et al. Front Immunol. 9:1090. 2018.
13. Vanderleyden I, Fra-Bido SC, Innocentin S, et al. Cell Rep. 30(3): 611–619.e4. 2020.
14. Bally AP, Tang Y, Lee JT, et al. J Immunol. 198(1):205–217. 2017.
15. Quatrini L, Wieduwild E, Escaliere B, et al. Nat Immunol. 19(9):954-962. 2018.
16. Shimizu K, Sugiura D, Okazaki IM, et al. Mol Cell. 77(5):937-950.e6. 2020.
17. Karnowski A, Chevrier S, Belz GT, et al. J Exp Med. 209(11):2049-2064. 2012.
18. Park HJ, Kusnadi A, Lee EJ, et al. Cell Immunol. 278(1-2):76-83. 2012.
19. Huang JR, Tsai YC, Chang YJ, et al. J Immunol. 192(4):1972-1981. 2014.
20. Park SJ, Namkoong H, Doh J, et al. J Leukoc Biol. 96(5):939. 2014.
21. Puleston DJ, Zhang H, Powell TJ, et al. Elife. 3:e03706. 2014.
22. Lu X, Ding ZC, Cao Y, et al. J Immunol. 194(4):2011-2021. 2015.
23. Bally AP, Lu P, Tang Y, et al. J Immunol. 194(9):4545-4554. 2015.
24. Zeng, W., Liu, Z., Zhang, S. et al. Sci Rep. 6:36560. 2016.
25. Zhuang Z, Lai X, Sun J, et al. J Exp Med. 218(4):e20202187. 2021.
26. Mitchell JE, Lund MM, Starmer J, et al. Cell Rep. 35(2):108966. 2021.
27. Christian LS, Wang L, Lim B, et al. Cell Rep. 35(6):109118. 2021.
28. Cui J, Xu H, Yu J, Ran S, Zhang X, et al. Sci Immunol. 9(94):eadh0085. 2024.
Depletion
Flow Cytometry
in vivo Protocol

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Disclaimer AlertProducts are for research use only. Not for use in diagnostic or therapeutic procedures.