Catalog No.	PA007163.r2a
Product Name	Anti-mouse PD-1 Monoclonal Antibody (Clone 29F.1A12.1) | PA007163.r2a
Supplier Name	Syd Labs, Inc.
Brand Name	Syd Labs
Synonyms	programmed cell death protein 1, PD-1, CD279, cluster of differentiation 279, 29F.1A12
Summary	The in vivo grade recombinant anti-mouse PD-1 / CD279 monoclonal antibody (Clone 29F.1A12.1, rat IgG2a kappa) was produced in mammalian cells.
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.
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.
Form Of Antibody	0.2 μM filtered solution of 1x PBS.
Endotoxin	Less than 1 EU/mg of protein as determined by LAL method.
Purity	>95% by SDS-PAGE under reducing conditions.
Shipping	The in vivo grade recombinant anti-mouse PD-1/CD279 monoclonal antibodies(Clone 29F.1A12.1) 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. 1 month from date of receipt, 2 to 8°C as supplied. 3 months from date of receipt, -20°C to -70°C as supplied.
Note	Recombinant anti-mouse PD1 monoclonal antibodies, which share the same variable region sequences with the rat anti-mouse PD-1 / CD279 monoclonal antibody (clone number: 29F.1A12), are produced from mammalian cells and good for in vitro and in vivo studies.
Order Offline	Phone: 1-617-401-8149 Fax: 1-617-606-5022 Email: message@sydlabs.com Or leave a message with a formal purchase order (PO) Or credit card.

Description

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

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(CD279) and anti-mouse PD-L1 antibodies(CD274) 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.

Syd Labs’s 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).

References for Anti-mouse PD-1 Antibody (Clone 29F.1A12.1):

1、RIP1 Kinase Drives Macrophage-Mediated Adaptive Immune Tolerance in Pancreatic Cancer.
Wang W, et al. Cancer Cell. 2018 Nov 12;34(5):757-774.e7. doi: 10.1016/j.ccell.2018.10.006. PMID: 30423296
“In other experiments animals were treated i.p. with Nec1s (2.5 mg/kg, daily; BioVision, Milpitas, CA), a neutralizing αPD-1 mAb (29F.1A12, 200 μg) or an agonizing ICOS mAb (7E.17G9, 100 μg, Days 4, 7 and 10 post-tumor challenge; both BioXcell) or respective isotype controls. …Pancreatic ductal adenocarcinoma (PDA) is characterized by immune tolerance and immunotherapeutic resistance. …Targeting RIP1 synergized with PD1-and inducible co-stimulator-based immunotherapies. …We recently reported GSK′963 as a potent and selective inhibitor of both murine and human RIP1; however, its low oral exposure makes it unsuitable for in vivo administration. …To further test whether the M1-like reprogramming of TAMs is responsible for the tumor-protective T cell phenotype in RIP1i-treated mice, we serially neutralized macrophages in PDA-bearing control and RIP1i-treated mice.”

2、PD-1 expression by tumour-associated macrophages inhibits phagocytosis and tumour immunity.
Gordon SR, et al. Nature. 2017 May 25;545(7655):495-499. doi: 10.1038/nature22396. PMID: 28514441
“Cytospinned TAMs were stained with fluorescently conjugated secondary antibodies only. n = 2, two experimental repeats. 20× magnification; scale bar, 20 μm. Red, 594; green, 488; blue, Hoechst. b, Mouse PD-1− TAMs trend towards an M1 (CD206−MHC IIhigh) expression profile, rather than M2 (CD206+MHC IIlow or negative). …Tumour cells frequently overexpress the ligand for PD-1, programmed cell death ligand 1 (PD-L1), facilitating their escape from the immune system. …Monoclonal antibodies that block the interaction between PD-1 and PD-L1, by binding to either the ligand or receptor, have shown notable clinical efficacy in patients with a variety of cancers, including melanoma, colorectal cancer, non-small-cell lung cancer and Hodgkin’s lymphoma. …Here we show that both mouse and human TAMs express PD-1. …TAM PD-1 expression correlates negatively with phagocytic potency against tumour cells, and blockade of PD-1–PD-L1 in vivo increases macrophage phagocytosis, reduces tumour growth and lengthens the survival of mice in mouse models of cancer in a macrophage-dependent fashion.”

3、STK11/LKB1 Deficiency Promotes Neutrophil Recruitment and Proinflammatory Cytokine Production to Suppress T-cell Activity in the Lung Tumor Microenvironment.
Koyama S, et al. Cancer Res. 2016 Mar 1;76(5):999-1008. doi: 10.1158/0008-5472.CAN-15-1439. PMID: 26833127
“Mice were dosed with 200 μg of IL6-neutralizing antibody (MP5-20F3, BioXcell), anti Ly-6G/Gr-1 antibody (RB6-8C5, BioXcell), PD-1–blocking antibody (clone 29F.1A12), and isotype controls (BioXcell) three times a week via intraperitoneal injections. …Furthermore, STK11/LKB1–inactivating mutations were associated with reduced expression of PD-1 ligand PD-L1 in mouse and patient tumors as well as in tumor-derived cell lines. …Our findings illustrate how tumor suppressor mutations can modulate the immune milieu of the tumor microenvironment, and they offer specific implications for addressing STK11/LKB1–mutated tumors with PD-1–targeting antibody therapies. …Recent clinical trials in NSCLC have demonstrated response to immune checkpoint blockade and nominated predictive markers for the efficacy of specific immunotherapies. …Together, the results suggest that in the Lkb1-deficient tumors, immune evasion is achieved through suppressive myeloid cells and aberrant cytokine production and not the PD-1:PD-L1 interaction.”

4、Adaptive resistance to therapeutic PD-1 blockade is associated with upregulation of alternative immune checkpoints.
Koyama S, et al. Nat Commun. 2016 Feb 17;7:10501. doi: 10.1038/ncomms10501. PMID: 26883990
“Anti-PD-1 therapeutic antibodies function through binding to PD-1 on tumour-reactive T cells and inhibiting the PD-1:PD-L1 interaction, thereby reinvigourating the anti-tumour T-cell response. …Conversely, immunotherapy approaches, specifically PD-1:PD-L1 blockade, appear to be broadly efficacious in NSCLC patients. …The EGFR and Kras models were treated with a therapeutic anti-PD-1 antibody until tumours demonstrated progression by magnetic resonance imaging (MRI) and evaluated immune profiles. …Despite compelling antitumour activity of antibodies targeting the programmed death 1 (PD-1): programmed death ligand 1 (PD-L1) immune checkpoint in lung cancer, resistance to these therapies has increasingly been observed. …In tumours progressing following response to anti-PD-1 therapy, we observe upregulation of alternative immune checkpoints, notably T-cell immunoglobulin mucin-3 (TIM-3), in PD-1 antibody bound T cells and demonstrate a survival advantage with addition of a TIM-3 blocking antibody following failure of PD-1 blockade.”

5、Response to BRAF inhibition in melanoma is enhanced when combined with immune checkpoint blockade.
Cooper, Z. A., et al. Cancer Immunol Res. 2014 Jul;2(7):643-54. doi: 10.1158/2326-6066.CIR-13-0215. PMID: 24903021
“Ipilimumab [a monoclonal antibody (mAb) targeting immunomodulatory CTLA4 receptor on T cells] was approved by the FDA recently based on a survival benefit over standard chemotherapy in patients with metastatic melanoma. …One exciting approach undergoing clinical investigation is the combination of BRAFi with immunotherapy to generate a sustained antitumor immune response. …BRAF-targeted therapy results in objective responses in the majority of patients; however, the responses are short lived (∼6 months). …There is also increased expression of the immunomodulatory molecule PDL1, which may contribute to the resistance. …Administration of anti-PD1 or anti-PDL1 together with a BRAF inhibitor led to an enhanced response, significantly prolonging survival and slowing tumor growth, as well as significantly increasing the number and activity of tumor-infiltrating lymphocytes.”

6、Negative role of inducible PD-1 on survival of activated dendritic cells.
Park, S. J., et al. J Leukoc Biol. 2014 Apr;95(4):621-9. doi: 10.1189/jlb.0813443. PMID: 24319287
“Recently, PD-1 was found to be expressed in innate cells, including activated DCs, and plays roles in suppressing production of inflammatory cytokines. …In this study, we demonstrate that PD-1 KO DCs exhibited prolonged longevity compared with WT DCs in the dLNs after transfer of DCs into hind footpads. Interestingly, upon LPS stimulation, WT DCs increased the expression of PD-1 and started to undergo apoptosis. …Moreover, treatment of blocking anti-PD-1 mAb during DC maturation resulted in enhanced DC survival, suggesting that PD-1:PD-L interactions are involved in DC apoptosis. …As a result, PD-1-deficient DCs augmented T cell responses in terms of antigen-specific IFN-γ production and proliferation of CD4 and CD8 T cells to a greater degree than WT DCs. …Taken together, our findings further extend the function of PD-1, which plays an important role in apoptosis of activated DCs and provides important implications for PD-1-mediated immune regulation.”

7、Dual blockade of PD-1 and CTLA-4 combined with tumor vaccine effectively restores T-cell rejection function in tumors.
Duraiswamy J, et al. Cancer Res. 2013 Jun;73(12):3591-603. doi: 10.1158/0008-5472.CAN-12-4100. PMID: 23633484
“Two hundred microgram of rat α-mouse PD-1 (29F.1A12), PD-L1 (10F.9G2), PD-L2 (3.2; ref. 24), or 100 μg of αCTLA-4 (clone 9D9) were administered intraperitoneally, either 3, 6, and 9 days following CT26 inoculation or 10, 13, and 16 days following ID8-VEGF inoculation. …Tumor progression is facilitated by regulatory T cells (Treg) and restricted by effector T cells. …In addition, we identify an additional role of CTL antigen-4 (CTLA-4) inhibitory receptor in further promoting dysfunction of CD8+ T effector cells in tumor models (CT26 colon carcinoma and ID8-VEGF ovarian carcinoma). …Double blockade was associated with increased proliferation of antigen-specific effector CD8+ and CD4+ T cells, antigen-specific cytokine release, inhibition of suppressive functions of Tregs, and upregulation of key signaling molecules critical for T-cell function. …Taken together, our study describes an approach involving vaccination and inhibitory receptor blockade to reverse T-cell exhaustion, which we envision to be a promising anticancer immunotherapy.”

8、CD80 expression on B cells regulates murine T follicular helper development, germinal center B cell survival, and plasma cell generation.
Good-Jacobson KL, et al. J Immunol. 2012 May 1;188(9):4217-25. doi: 10.4049/jimmunol.1102885. PMID: 22450810
“Because CD80 is one of the few markers shared by human and murine memory B cells, we investigated its role in the development of GCs, memory cells, and PCs. …In CD80-deficient mice, fewer long-lived PCs were generated upon immunization compared with that in B6 controls. …In concert, the absence of CD80 resulted in an increase in apoptotic GC B cells during the contraction phase of the GC. CD80−/− mice had fewer TFH cells compared with that of B6, and residual TFH cells failed to mature, with decreased ICOS and PD-1 expression and decreased synthesis of IL-21 mRNA. …Mixed bone marrow chimeras demonstrated a B cell-intrinsic requirement for CD80 expression for normal TFH cell and PC development. …These data provide new insights into the development of GCs and Ab-forming cells and the functions of CD80 in humoral immunity.”

9、Role of the immune modulator programmed cell death-1 during development and apoptosis of mouse retinal ganglion cells.
Sham CW, Chan AM, Francisco LM, et al. Invest Ophthalmol Vis Sci. 2009 Oct;50(10):4941-8. doi: 10.1167/iovs.09-3602. PMID: 19420345
“For the antibody blocking experiment, the following functional grade antibodies were used at a final concentration of 7 μg/mL: anti–mouse PD-1 (clone J43, hamster IgG; eBioscience, San Diego, CA) for which specificity 12 and ability to block interaction with PD-L1 and PD-L2 13 have previously been described, or isotype control hamster IgG (eBioscience). …Retinal sections were imaged at 200× magnification with a microscope (E800; Nikon, Tokyo, Japan) and a digital camera (SPOTII; Diagnostic Instruments, Sterling Heights, MI), yielding a retinal image width of 0.63 mm. …In the adult retina, PD-1 expression was detected primarily in the GCL, and a subpopulation of cells was detected in the inner nuclear layer. …Mammalian programmed cell death (PD)-1 is a membrane-associated receptor regulating the balance between T-cell activation, tolerance, and immunopathology; however, its role in neurons has not yet been defined. …Mature retinal cell types expressing PD-1 were identified by immunofluorescence staining of vertical retina sections; developmental expression was localized by immunostaining and quantified by Western blot analysis.”

10、Restoring function in exhausted CD8 T cells during chronic viral infection.
Barber, D. L., et al. Nature. 2006 9;439(7077):682-7. doi: 10.1038/nature04444. PMID: 16382236
“Functional impairment of antigen-specific T cells is a defining characteristic of many chronic infections, but the underlying mechanisms of T-cell dysfunction are not well understood. …To address this question, we analysed genes expressed in functionally impaired virus-specific CD8 T cells present in mice chronically infected with lymphocytic choriomeningitis virus (LCMV), and compared these with the gene profile of functional memory CD8 T cells. …Here we report that PD-1 (programmed death 1; also known as Pdcd1) was selectively upregulated by the exhausted T cells, and that in vivo administration of antibodies that blocked the interaction of this inhibitory receptor with its ligand, PD-L1 (also known as B7-H1), enhanced T-cell responses. …Notably, we found that even in persistently infected mice that were lacking CD4 T-cell help, blockade of the PD-1/PD-L1 inhibitory pathway had a beneficial effect on the ‘helpless’ CD8 T cells, restoring their ability to undergo proliferation, secrete cytokines, kill infected cells and decrease viral load. Blockade of the CTLA-4 (cytotoxic T-lymphocyte-associated protein 4) inhibitory pathway had no effect on either T-cell function or viral control. …These studies identify a specific mechanism of T-cell exhaustion and define a potentially effective immunological strategy for the treatment of chronic viral infections.”

Related Recombinant IgG Reference Antibodies:
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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, In vivo Grade

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

Anti-mouse PD-1 Monoclonal Antibody(29F.1A12.1) from: Anti-mouse PD-1 Monoclonal Antibody (Clone 29F.1A12.1): PA007163.r2a Syd Labs

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