Anti-Mouse CD279 (PD-1) [Clone 29F.1A12] — Purified in vivo GOLD™ Functional Grade
Anti-Mouse CD279 (PD-1) [Clone 29F.1A12] — Purified in vivo GOLD™ Functional Grade
Product No.: P377
Clone 29F.1A12 Target PD-1 Formats AvailableView All Product Type Monoclonal Antibody Alternate Names Programmed Death-1, CD279 Isotype Rat IgG2a Applications B , CyTOF® , FC , IHC FF , in vivo , PhenoCycler® , WB |
Data
Antibody DetailsProduct DetailsReactive Species Mouse Host Species Rat Recommended Isotype Controls Recommended Isotype Controls Recommended Dilution Buffer Immunogen PD-1 cDNA followed by PD-1-Ig fusion protein Product Concentration ≥ 5.0 mg/ml Endotoxin Level < 1.0 EU/mg 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 RRIDAB_2737558 Additional Applications Reported In Literature ? CyTOF® Each investigator should determine their own optimal working dilution for specific applications. See directions on lot specific datasheets, as information may periodically change. DescriptionDescriptionSpecificity Clone 29F.1A12 recognizes an epitope on mouse PD-1. Background PD-1 is a 50-55 kD member of the B7 Ig superfamily. PD-1 is also a member of the extended CD28/CTLA-4 family of T cell regulators and is suspected to play a role in lymphocyte clonal selection and peripheral tolerance. The ligands of PD-1 are PD-L1 and PD-L2, and are also members of the B7 Ig superfamily. PD-1 and its ligands negatively regulate immune responses. PD-L1, or B7-Homolog 1, is a 40 kD type I transmembrane protein that has been reported to costimulate T cell growth and cytokine production. The interaction of PD-1 with its ligand PD-L1 is critical in the inhibition of T cell responses that include T cell proliferation and cytokine production. PD-L1 has increased expression in several cancers. Inhibition of the interaction between PD-1 and PD-L1 can serve as an immune checkpoint blockade by improving T-cell responses In vitro and mediating preclinical antitumor activity. Within the field of checkpoint inhibition, combination therapy using anti-PD1 in conjunction with anti-CTLA4 has significant therapeutic potential for tumor treatments. PD-L2 is a 25 kD type I transmembrane ligand of PD-1. Via PD-1, PD-L2 can serve as a co-inhibitor of T cell functions. Regulation of T cell responses, including enhanced T cell proliferation and cytokine production, can result from mAbs that block the PD-L2 and PD-1 interaction. Antigen Distribution PD-1 is expressed on a subset of CD4-CD8- thymocytes, and on activated T and B cells. Ligand/Receptor B7-H1 (PD-L1) and B7-DC (PD-L2) Function Lymphocyte clonal selection, peripheral tolerance NCBI Gene Bank ID UniProt.org Research Area Cancer . Immunology . Inhibitory Molecules Leinco Antibody AdvisorPowered 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 29F.1A12 is a rat anti-mouse PD-1 monoclonal antibody widely used in in vivo mouse studies to block the interaction between PD-1 and its ligands, typically as a tool to study immune checkpoint blockade therapy. In these studies, 29F.1A12 is used to:
Typical experimental protocols include:
Considerations:
In summary, clone 29F.1A12 is a gold-standard tool for in vivo PD-1 blockade in mice, supporting functional studies of immune modulation, cancer immunotherapy, and basic T-cell biology. The correct storage temperature for sterile packaged clone 29F.1A12 is 2 to 8°C as supplied; it must not be frozen. Further details:
If your product is a sterile, working solution packaged for research (not an unconjugated purified bulk for long-term archive), use 2–8°C storage conditions. Commonly Used Antibodies and Proteins Co-Used with 29F.1A12 in Literature29F.1A12 is a monoclonal antibody targeting mouse PD-1, commonly used in research to block the PD-1/PD-L1 pathway and study its role in immune regulation and cancer biology. Several other antibodies and proteins are frequently used alongside 29F.1A12 in experimental setups, especially in flow cytometry, in vitro blocking assays, and co-staining experiments. Common Companion AntibodiesCo-Staining with Other PD-1 Antibodies
Blocking PD-L1 Interactions
Co-Use with Anti-PD-L1 Antibodies
Typical Experimental Approaches
SummaryIn the literature, 29F.1A12 is most frequently used alongside other anti-PD-1 antibodies like RMP1-30 (for co-staining and validation) and RMP1-14 (for comparative blocking studies). Functional experiments often include recombinant PD-L1 to assess blockade efficacy, while parallel studies may use anti-PD-L1 antibodies such as 10F.9G2 and MIH6 to model comprehensive pathway inhibition. These combinations are standard in preclinical research aiming to dissect the PD-1/PD-L1 axis and evaluate potential therapeutic interventions. The 29F.1A12 anti-PD-1 antibody clone has emerged as a significant research tool in immunotherapy studies, with several key findings documented across scientific literature. Specificity and Detection PropertiesThe 29F.1A12 clone demonstrates robust specificity for mouse PD-1, recognizing a specific epitope on the PD-1 protein. Flow cytometric analyses have shown that this clone effectively detects PD-1 surface protein expression on live murine cells, with particularly high reactivity observed on activated T-cells (52.2 ± 7.9%) compared to unactivated T-cells (6.0 ± 1.5%). Importantly, the clone shows minimal cross-reactivity with PD-1 knockout cells, confirming its specificity for live cells while showing lesser specificity for dead cells. Functional Blocking ActivityA critical finding is that 29F.1A12 functions as a blocking antibody that prevents PD-1 from interacting with its ligand PD-L1. This blocking capability is so comprehensive that it completely prevents PD-1 detection by nearly all other antibody clones when used in competition assays. The antibody's blocking function translates into therapeutic potential, as it can inhibit B16-F10 melanoma growth in three-dimensional tumor spheroid cultures, though notably not in standard two-dimensional cultures. Clone Comparison and OverlapWhen compared to other anti-PD-1 clones like RMP1-30, the 29F.1A12 antibody shows the brightest staining intensity among tested clones. Co-staining experiments reveal that 29F.1A12 and RMP1-30 recognize overlapping cell subpopulations, with dual positivity observed in nearly all PD-1-reactive melanoma cells (90.8-96.7%) and T-cells (48.1-57.2%). Culture Condition SensitivityA notable finding is that both 29F.1A12 and RMP1-30 reactivity increases more than 3-fold in three-dimensional versus two-dimensional culture conditions. This suggests that PD-1 expression levels vary significantly depending on the cellular environment, which has important implications for experimental design and therapeutic applications. Variable Therapeutic EfficacyIn preclinical studies, 29F.1A12 has shown differential efficacy across genetic contexts. While the clone demonstrated initial benefits in delaying cancer onset and improving survival in certain mouse models (such as Pole L424V mutant mice), it showed variable or limited effects in other genetic backgrounds compared to alternative clones like RMP-14. This variability highlights the importance of clone selection based on specific experimental contexts and genetic models. Technical ConsiderationsAn important caveat identified in the literature is that PD-1-specific blocking antibodies like 29F.1A12 can deplete PD-1+ T cells, presenting a potential confounding variable in preclinical immunotherapy experiments. This depletion effect must be considered when interpreting experimental results and designing studies. The collective findings establish 29F.1A12 as a highly specific, functionally active anti-PD-1 antibody with strong blocking capabilities, but researchers should carefully consider its variable efficacy across different experimental contexts and potential for T-cell depletion when designing studies. References & Citations1.) Ardolino, M. et al. (2018) J Clin Invest. 128(10):4654-4668. PubMed 2.) Schreiber, RD. et al. (2017) Cancer Immunol Res. 5(2):106-117. 3.) Honjo, T. et al. (1992) EMBO J. 11:3887. 4.) Wurster S. et al. (2020) The Journal of Infectious Diseases 222(6):1989–994 Journal Link 5.) Lo, R. et al. (2021) Cancer Cell 39(10):1375-1387.e6 Journal Link Technical ProtocolsCertificate of Analysis |
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P377 | |
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P378 |
