Anti-Mouse PD-1 Monoclonal Antibodies

Background

The rat anti-mouse PD-1 monoclonal antibody 29F.1A12  (rat IgG2a kappa) reacts with the mouse PD-1 protein (programmed death-1 or CD279) encoded by the mouse Pdcd1 gene, a member of the CD28 family of the Ig superfamily. PD-1 has two ligands, PD-L1 and PD-L2, both of which belong to the B7 family. It has been shown that in mouse models of melanoma, tumor growth can be transiently arrested via treatment with the anti-mouse PD-1 and anti-mouse PD-L1 antibodies which block the interaction between the PD-L1 protein and its receptor PD-1 protein. The 29F.1A12 monoclonal antibody blocks the binding of both the mouse PD-L1 protein and the mouse PD-L2 protein to the mouse PD-1 protein.

Our recombinant 29F.1A12 antibodies have a part (variable regions) or complete amino acid sequences of the rat anti-mouse PD-1 monoclonal antibody (hybridoma clone name or number: 29F.1A12).

PA007163.m2cLA: In Vivo Grade Recombinant Anti-mouse PD-1 Mouse IgG2c-L234A L235A P329G (LALAPG) Kappa Monoclonal Antibody (Clone 29F.1A12.1)

The in vivo grade recombinant anti-mouse PD-1 / CD279 monoclonal antibody (mouse IgG2c-LALAPG kappa) was produced in mammalian cells.
Immunogen: The original rat hybridoma (clone name: 29F.1A12) was generated by immunizing rats with the mouse PD-1 cDNA followed by the mouse PD-1-Ig fusion protein.
Clone: 29F.1A12.1, the same variable region sequences as the rat anti-mouse PD-1 monoclonal antibody (clone number: 29F.1A12).
Isotype: mouse IgG2c, kappa.
Applications: western blot (WB), immunohistochemistry (IHC), Flow Cytometry (FC), and various in vitro and in vivo functional assays.
Formulation: 0.2 μM filtered solution of 1x PBS.
Purity: >95% by SDS-PAGE under reducing conditions.
Endotoxin Level: Less than 1 EU/mg of protein as determined by LAL method.

PA007163.r2a: In Vivo Grade Recombinant Anti-mouse PD-1 Rat IgG2a Kappa Monoclonal Antibody (Clone 29F.1A12.1)

The in vivo grade recombinant anti-mouse PD-1 / CD279 monoclonal antibody (rat IgG2a kappa) was produced in mammalian cells. Its affinity to the mouse PD1 protein is <20 nM.
Immunogen: The original rat hybridoma (clone name: 29F.1A12) was generated by immunizing rats with the PD-1 cDNA followed by the PD-1-Ig fusion protein.
Clone: 29F.1A12.1, the same variable region and constant region sequences as the rat anti-mouse PD-1 monoclonal antibody (clone number: 29F.1A12).
Isotype: rat IgG2a, kappa.
Applications: western blot (WB), immunohistochemistry (IHC), Flow Cytometry (FC), and various in vitro and in vivo functional assays.
Formulation: 0.2 μM filtered solution of 1x PBS.
Purity: >95% by SDS-PAGE under reducing conditions.
Endotoxin Level: Less than 1 EU/mg of protein as determined by LAL method.

Shipping: The in vivo grade recombinant anti-mouse PD-1 / CD279 monoclonal antibodies are shipped with ice pack. Upon receipt, store it immediately at the temperature recommended below.
Stability & Storage: Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
3 month from date of receipt, 2 to 8°C as supplied.
12 months from date of receipt, -20°C to -70°C as supplied.

References of anti-mouse PD-1 antibody (29F.1A12)

An affinity threshold for maximum efficacy in anti-PD-1 immunotherapy.
Cowles, S. C., et al. mAbs. 2022;14(1):2088454. doi: 10.1080/19420862.2022.2088454. PMID: 35924382
We started with a parental anti-PD-1 antibody clone, 29F.1A12 (anti-mouse PD-1 antibody). The murine PD-1 monoFc fusion protein was also validated by enzyme-linked immunosorbent assay (ELISA) to commercially purchased 29F.1A12 antibody (anti-mouse PD-1 mAb). A DNA sequence encoding the variable light and heavy fragments of the anti-PD-1 antibody clone 29F.1A12 (anti-mouse PD-1/CD279 antibody) were purchased as a GeneBlock (IDT) and cloned using the same reagents as above in the pCTCON2 vector. The antibody clone 29F.1A12 (anti-mouse PD-1 mAb) was expressed on the surface of yeast in the scFv format as a fusion protein to Aga2p. We used a yeast display library to identify variants of 29F.1A12 with improved binding affinity to PD-1.
Tags: anti-mouse PD-1 antibody (29F.1A12); 29F.1A12 antibody for cancer research

Immunogenicity and antitumor efficacy of a novel human PD-1 B-cell vaccine (PD1-Vaxx) and combination immunotherapy with dual trastuzumab/pertuzumab-like HER-2 B-cell epitope vaccines (B-Vaxx) in a syngeneic mouse model.
Kaumaya, P. T. P., et al. OncoImmunology. 2020;9(1):1818437. doi: 10.1080/2162402X.2020.1818437. PMID: 33117602
Mice were treated twice a week with anti-mPD-1 mAb 29F.1A12 (anti-mouse PD-1 antibody) 200 µg/dose reconstituted in PBS was used as a positive control. Control mice were treated twice weekly with PBS as negative control and twice weekly with anti-mPD-1 mAb 29F.1A12 (anti-mouse PD-1 mAb) as positive control starting 2 days after tumor challenge. Negative control mice were treated with PBS twice weekly and positive control group were treated with anti-mouse PD-1mAb 29F.1A12 (anti-mouse PD-1/CD279 antibody) twice weekly after challenged with 10^5 CT26 tumor cells. Control mice were treated twice weekly with PBS as negative control and twice weekly with anti-mPD-1 mAb (29F.1A12) as positive control starting at Day 2 after challenge. The combination of PD1-Vaxx with B-Vaxx showed significant tumor growth inhibition compared to 29F.1A12 alone.
Tags: activity of 29F.1A12 antibody; in vivo study of 29F.1A12 antibody

PD-1-specific "Blocking" antibodies that deplete PD-1+ T cells present an inconvenient variable in preclinical immunotherapy experiments.
Polesso, F., et al. Eur J Immunol. 2021;51(6):1460-1470. doi: 10.1002/eji.202048960. PMID: 33684223
29F.1A12 (anti-mouse PD-1 monoclonal antibody) staining was brightest, RMP1-14 staining was weakest, while J43, G4 and RMP1-30 each stained with intermediate intensity. 29F.1A12 antibody (anti-mouse PD-1 mAb) is reported to prevent PD-1 from interacting with PD-L1, and these data show that it also completely prevents PD-1 detection with nearly all other clones. Blocking was performed using unconjugated 29F.1A12 (anti-mouse PD-1/CD279 antibody), unconjugated J43, unconjugated G4, unconjugated RMP1-14, or unconjugated RMP1-30. Staining was performed using PE-labeled 29F.1A12 antibody (anti-mouse PD-1 mAb), PE-labeled J43, PE-labeled RMP1-14, PE-labeled RMP1-30, or unconjugated G4 followed by PE-conjugated mouse anti-hamster IgG cocktail. These findings highlight the variability in PD-1 detection due to antibody clone interactions.
Tags: 29F.1A12 antibody; 29F.1A12 antibody mouse dose range

Immune Checkpoint Blockade Delays Cancer and Extends Survival in Murine DNA Polymerase Mutator Syndromes.
Sawant, A., et al. bioRxiv. 2024. doi: 10.1101/2024.06.10.597960. PMID: 38915517
Two clones of anti-PD1 antibodies- RMP-14 and 29F.1A12 (anti-mouse PD-1 antibody) clones were used for biweekly treatments in all Pold1 and Pole proofreading mutation mouse models. The same experiment was performed with 29F.1A12 clone of anti-PD1 antibody (anti-mouse PD-1 mAb), however, a significant improvement in survival as was seen with the RMP-14 clone was not observed. In contrast to the Pold1 D400A mouse model, we observed that anti-PD1 antibody treatment with the 29F.1A12 clone (anti-mouse PD-1/CD279 antibody) in Pole L424V mutant mice initially delayed cancer onset and improved survival compared to the RMP-14 clone treated cohort. These findings suggest differential efficacy of anti-PD-1 clones in specific genetic contexts. Anti-PD-1 treatment with 29F.1A12 showed variable effects across different mouse models.
Tags: 29F.1A12 mAb; 29F.1A12 monoclonal antibody

Anti–PD-1 antibodies recognizing the membrane-proximal region are PD-1 agonists that can down-regulate inflammatory diseases.
Suzuki, K., et al. Sci Immunol. 2023;8(79):eadd4947. doi: 10.1126/sciimmunol.add4947. PMID: 36638191
Anti-mouse PD-1 (mPD-1) mAb clones, 29F.1A12 and RMP1-14, were coimmobilized with anti-CD3 and anti-CD28 mAbs, but neither of these anti-PD-1 mAbs (29F.1A12, anti-mouse PD-1 antibody) could inhibit T cell activity. Simple coligation of TCR and PD-1 using arbitrary antibodies failed to trigger PD-1 stimulation, suggesting the presence of specific epitope recognition requirements for functional anti-PD-1 agonist antibodies (29F.1A12, anti-mouse PD-1 mAb). Given that both 29F.1A12 (anti-mouse PD-1/CD279 antibody) and RMP1-14 are blocking antibodies, we suspected that anti-PD-1 agonistic mAbs might be recognizing a separate epitope from the ligand-binding domain. These findings indicate that 29F.1A12 may not function as a PD-1 agonist in certain contexts. The study highlights the complexity of PD-1 signaling modulation by different antibody clones.
Tags: anti-mouse PD-1 antibody (clone 29F.1A12); clone 29F.1A12 of anti-mouse PD-1

For more references about anti-mouse PD-1 antibody (clone 29F.1A12), please contact our scientific support team with message@sydlabs.com.

Related Recombinant IgG Reference Antibodies:
In vivo Grade Recombinant Mouse IgG1 Isotype Control Antibody and Mutants
In vivo Grade Recombinant Mouse IgG2a Isotype Control Antibody and Mutants
In vivo Grade Recombinant Mouse IgG2c Isotype Control Antibody and Mutants
In vivo Grade Recombinant Rat IgG2a Isotype Control Antibody

Syd Labs provides the following anti-mouse PD-L1 / PD-1 antibodies:
In vivo grade recombinant anti-mouse PD1 antibodies (Clone 29F.1A12.1)
In vivo grade recombinant anti-mouse PD-1 antibodies (Clone RMP1-14.1)
In vivo grade recombinant anti-mouse PD-L1 antibodies (Clone 10F.9G2.1)
In vivo grade recombinant anti-mouse PD-1 / PD-1 bispecific antibodies (Clone 29F.1A12.1 / RMP1-14.1)
In vivo grade recombinant anti-mouse PD-1 / PD-L1 bispecific antibodies (Clone 29F.1A12.1 / 10F.9G2.1)

Recombinant PD1 and PDL1 Proteins:
Recombinant Human PD-1/PDCD1 Proteins
Biotinylated Recombinant Human PD-1/PDCD1 Proteins
Recombinant Mouse PD-1/PDCD1 Proteins
Biotinylated Recombinant Mouse PD-1/PDCD1 Proteins
Recombinant Cynomolgus PD-1/PDCD1 Proteins
Recombinant Canine PD-1/PDCD1 Proteins
Biotinylated Cynomolgus/Rhesus macaque PD-L1 Protein
Biotinylated Human PD-L1 Protein
Biotinylated Mouse PD-L1 Protein
Cynomolgus PD-L1 Protein
Cynomolgus/Rhesus macaque PD-L1 Protein
Human PD-L1/B7-H1 Protein
Mouse PD-L1/B7-H1 Protein

Questions and Answers about anti-mouse PD-1 antibody (Clone 29F.1A12):

Question: Which anti-mouse PD1 antibody works the best?
Answer: (by AI) There are several anti-mouse PD1 antibodies available in the market. Bio X Cell offers three different anti-mouse PD-1 monoclonal antibodies: Clones RMP1-14, 29F.1A12, and J43. All three of these antibodies are commonly used to block PD-1 signaling in vivo in murine tumor models and other mouse models. These three clones all have extensive multi-year publication records supporting them. Leinco Technologies offers four different anti-PD-1 monoclonal antibodies: 29F.1A12, RMP1-14, J43, and 5C4-B83. The RMP1-14 antibody has been reported to block the binding of PD-1 to its ligands (B7-H1 and B7-DC) and to inhibit T cell proliferation and cytokine production costimulated by macrophages (but not by dendritic cells and B cells). (by Syd Labs) Syd Labs offers anti-mouse PD-1 / CD279 monoclonal antibodies based on the sequences of clones RMP1-14 and 29F.1A12. Syd Labs provides in vivo grade recombinant antibodies including engineered antibodies for the clones RMP1-14 and 29F.1A12.

Question: Do you produce Fc-silenced 29F.1A12 antibody?
Answer: We provide recombinant 29F.1A12 antibodies including the Fc-silenced or Fc silent versions of recombinant 29F.1A12 antibodies. It is very easy for us to produce the Fc-silenced or Fc silent versions of recombinant 29F.1A12 antibodies no matter which isotypes they are based, for example, mIgG2c LALAPG.

Question: What are the published applications of the 29F.1A12 antibody?
Answer: It was reported that the blockade of the PD-1 protein expressed on TME-infiltrating macrophages and natural killer (NK) cells by 29F.1A12 results in tumor growth suppression and prolonged survival. It is also reported that the 29F.1A12 antibody recognizes the PD-1 surface protein on B16-F10 and other murine tumor lines, suppresses cancer cell-intrinsic PD-1 receptor signaling, and inhibits resultant tumor growth in three-dimensional (3D) culture and tumor-bearing mice. In the paper "Distinct antibody clones detect PD-1 checkpoint expression and block PD-L1 interactions on live murine melanoma cells", it was shown that the anti-PD-1 29F.1A12 monoclonal antibody recognizes the PD-1 surface protein on live B16-F10 melanoma cells, live YUMM melanoma variant lines, and overlapping WT murine B16-F10 and YUMM melanoma and T-cell subsets by FACS. In contrast to the RMP1-30 antibody, The 29F.1A12 PD-1 blocking antibody inhibits PD-1 interactions with PD-L1 on live B16-F10 melanoma cells. The RMP1-14 clone, but not 29F.1A12 or RMP1-30, is useful for pulling down (IP) and sequencing PD-1 from B16-F10 and T-cell cultures, consistent with the previous observations that individual antibodies differ in their suitability for specific assay types. 29F.1A12 binding to B16-F10 cells was increased in 3D versus 2D settings.