Mouse IFN-gamma ELISpot Kit Summary
* Provided that the recommended microplates, buffers, diluents, substrates and solutions are used, and the assay is run as summarized in the Assay Procedure provided.
PRODUCT SUMMARY
ELISpot kits are highly sensitive, microplate-based assays for the detection of cytokine secreting cells. This kit is designed for the detection and enumeration of mouse IFN-gamma. Complete ELISpot kits are ready-to-run and require no assay development or refinement.
This ELISpot assay employs a capture antibody specific for mouse IFN-gamma, pre-coated onto a PVDF-backed microplate. Appropriately stimulated cells are pipetted directly into the wells and the immobilized antibody in the immediate vicinity of the secreting cells binds secreted mouse IFN-gamma. Following wash steps and incubation with a biotinylated detection antibody, alkaline-phosphatase conjugated to streptavidin is added. Unbound enzyme is subsequently removed by washing and a substrate solution (BCIP/NBT) is added. A blue-black colored precipitate forms at the sites of cytokine localization and appears as spots, with each individual spot representing an individual mouse IFN-gamma secreting cell. The spots can be counted with an automated ELISpot reader system or manually using a stereomicroscope.
PRODUCT FEATURES
- Detect and quantitate individual cells secreting mouse IFN-gamma
- High sensitivity - ELISpot assays can measure responses with frequencies well below 1 in 100,000 cells
- No in vitro expansion of cells required
- High-throughput - ELISpot assays use only a small number of primary cells
KIT CONTENTS
- Mouse IFN-gamma Microplate
- Biotinylated Detection Antibody
- Streptavidin conjugated to Alkaline Phosphatase
- Dilution Buffers
- Wash Buffer Concentrate
- BCIP/NBT Chromogen
- Mouse IFN-gamma Positive Control
OTHER REAGENTS REQUIRED
- Pipettes and pipette tips
- Deionized or distilled water
- Squirt bottle, manifold dispenser, or automated microplate washer
- 500 mL graduated cylinder
- 37 °C CO2 incubator
- Sterile culture media
- Dissection microscope or an automated ELISpot reader
Product Datasheets
Preparation and Storage
Background: IFN-gamma
IFN-gamma (Interferon-gamma) is the prototype proinflammatory cytokine and is produced by a variety of immune cells under inflammatory conditions, notably by T cells and NK cells. It plays a key role in host defense by promoting the development and activation of Th1 cells, chemoattraction and activation of monocytes and macrophages, upregulation of antigen presentation molecules, and immunoglobulin class switching in B cells. It also exhibits antiviral, antiproliferative, and apoptotic effects. In addition, IFN-gamma functions as an anti-inflammatory mediator by promoting the development of regulatory T cells and inhibiting Th17 cell differentiation. IFN-gamma dimers signal through a receptor complex of two IFN-gamma R1 and two IFN-gamma R2 subunits.
Citations for Mouse IFN-gamma ELISpot Kit
R&D Systems personnel manually curate a database that contains references using R&D Systems products. The data collected includes not only links to publications in PubMed, but also provides information about sample types, species, and experimental conditions.
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Citations: Showing 1 - 10
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Microbiota-induced plastic T cells enhance immune control of antigen-sharing tumors
Authors: Najar, TA;Hao, Y;Hao, Y;Romero-Meza, G;Dolynuk, A;Littman, DR;
bioRxiv : the preprint server for biology
Species: Mouse
Sample Types: Whole Cells
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Zeolitic imidazolate frameworks activate endosomal Toll-like receptors and potentiate immunogenicity of SARS-CoV-2 spike protein trimer
Authors: Alsaiari, SK;Nadeef, S;Daristotle, JL;Rothwell, W;Du, B;Garcia, J;Zhang, L;Sarmadi, M;Forster, TA;Menon, N;Lin, SQ;Tostanoski, LH;Hachmann, N;Wang, EY;Ventura, JD;Barouch, DH;Langer, R;Jaklenec, A;
Science advances
Species: Mouse
Sample Types: Whole Cells
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mRNAs encoding self-DNA reactive cGAS enhance the immunogenicity of lipid nanoparticle vaccines
Authors: Zhivaki, D;Gosselin, EA;Sengupta, D;Concepcion, H;Arinze, C;Chow, J;Nikiforov, A;Komoroski, V;MacFarlane, C;Sullivan, C;Kagan, JC;
mBio
Species: Mouse
Sample Types: Whole Cells
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Activation of human STING by a molecular glue-like compound
Authors: Li, J;Canham, SM;Wu, H;Henault, M;Chen, L;Liu, G;Chen, Y;Yu, G;Miller, HR;Hornak, V;Brittain, SM;Michaud, GA;Tutter, A;Broom, W;Digan, ME;McWhirter, SM;Sivick, KE;Pham, HT;Chen, CH;Tria, GS;McKenna, JM;Schirle, M;Mao, X;Nicholson, TB;Wang, Y;Jenkins, JL;Jain, RK;Tallarico, JA;Patel, SJ;Zheng, L;Ross, NT;Cho, CY;Zhang, X;Bai, XC;Feng, Y;
Nature chemical biology
Species: Mouse
Sample Types: Whole Cells
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The role of zinc sulfate in enhancing cellular and humoral immune responses to foot-and-mouth disease vaccine
Authors: Ko, MK;Kim, HW;Park, SH;Park, JH;Kim, SM;Lee, MJ;
Virus research
Species: Mouse
Sample Types: Cell Culture Supernates
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Killing tumor-associated bacteria with a liposomal antibiotic generates neoantigens that induce anti-tumor immune responses
Authors: Wang, M;Rousseau, B;Qiu, K;Huang, G;Zhang, Y;Su, H;Le Bihan-Benjamin, C;Khati, I;Artz, O;Foote, MB;Cheng, YY;Lee, KH;Miao, MZ;Sun, Y;Bousquet, PJ;Hilmi, M;Dumas, E;Hamy, AS;Reyal, F;Lin, L;Armistead, PM;Song, W;Vargason, A;Arthur, JC;Liu, Y;Guo, J;Zhou, X;Nguyen, J;He, Y;Ting, JP;Anselmo, AC;Huang, L;
Nature biotechnology
Species: Mouse
Sample Types: Whole Cells
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A SARS-CoV-2 and influenza double hit vaccine based on RBD-conjugated inactivated influenza A virus
Authors: Wang, Z;Li, Z;Shi, W;Zhu, D;Hu, S;Dinh, PC;Cheng, K;
Science advances
Species: Mouse
Sample Types: Whole Cells
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Monocyte production of C1q potentiates CD8 + T cell effector function following respiratory viral infection
Authors: Eddens, T;Parks, OB;Lou, D;Fan, L;Sojati, J;Ramsey, MJ;Schmitt, L;Salgado, CM;Reyes-Mugica, M;Oury, TD;Byersdorfer, C;Chen, K;Williams, JV;
bioRxiv : the preprint server for biology
Species: Mouse, Transgenic Mouse
Sample Types: Whole Cells
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Surface-modified measles vaccines encoding oligomeric, fusion-stabilized SARS-CoV-2 spike glycoproteins bypass measles seropositivity, boosting neutralizing antibody responses to omicron and historical variants
Authors: MÁ Muñoz-Alía, RA Nace, B Balakrishn, L Zhang, N Packiriswa, G Singh, P Warang, I Mena, R Narjari, R Vandergaas, A García-Sas, M Schotsaert, SJ Russell
bioRxiv : the preprint server for biology, 2022-12-16;0(0):.
Species: Mouse
Sample Types: Whole Cells
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Suppression of pullulanase-induced cytotoxic T cell response with a dual promoter in GSD IIIa mice
Authors: JA Lim, PS Kishnani, B Sun
JCI Insight, 2022-12-08;7(23):.
Species: Mouse
Sample Types: Tissue Homgenates
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Enhancing the Immunogenicity of Vaccinia Virus
Authors: SN Shchelkuno, SN Yakubitski, AA Sergeev, EV Starostina, KA Titova, SA Pyankov, GA Shchelkuno, MB Borgoyakov, AM Zadorozhny, LA Orlova, DN Kisakov, LI Karpenko
Viruses, 2022-06-30;14(7):.
Species: Mouse
Sample Types: Cell Culture Supernates
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Cure of syngeneic carcinomas with targeted IL-12 through obligate reprogramming of lymphoid and myeloid immunity
Authors: Y Hong, Y Robbins, X Yang, WK Mydlarz, A Sowers, JB Mitchell, JL Gulley, J Schlom, SR Gameiro, C Sievers, CT Allen
JCI Insight, 2022-03-08;7(5):.
Species: Mouse
Sample Types: Whole Cells
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Disruption of the crypt niche promotes outgrowth of mutated colorectal tumor stem cells
Authors: S Klingler, KS Hsu, G Hua, ML Martin, M Adileh, T Baslan, Z Zhang, PB Paty, Z Fuks, AM Brown, R Kolesnick
JCI Insight, 2022-03-08;7(5):.
Species: Mouse
Sample Types: Whole Cells
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Self-Assembled Particles Combining SARS-CoV-2 RBD Protein and RBD DNA Vaccine Induce Synergistic Enhancement of the Humoral Response in Mice
Authors: MB Borgoyakov, LI Karpenko, AP Rudometov, EA Volosnikov, IA Merkuleva, EV Starostina, AM Zadorozhny, AA Isaeva, VS Nesmeyanov, DV Shanshin, KO Baranov, NV Volkova, BN Zaitsev, LA Orlova, AV Zaykovskay, OV Pyankov, ED Danilenko, SI Bazhan, DN Shcherbako, AV Taranin, AA Ilyichev
International Journal of Molecular Sciences, 2022-02-16;23(4):.
Species: Mouse
Sample Types: Whole Cells
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In situ vaccination using unique TLR9 ligand K3-SPG induces long-lasting systemic immune response and synergizes with systemic and local immunotherapy
Authors: H Okada, K Takahashi, H Yaku, K Kobiyama, K Iwaisako, X Zhao, M Shiokawa, N Uza, Y Kodama, KJ Ishii, H Seno
Scientific Reports, 2022-02-08;12(1):2132.
Species: Mouse
Sample Types: Whole Cells
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Disruption of the crypt niche promotes outgrowth of mutated colorectal tumor stem cells
Authors: S Klingler, KS Hsu, G Hua, ML Martin, M Adileh, T Baslan, Z Zhang, PB Paty, Z Fuks, AM Brown, R Kolesnick
JCI Insight, 2022;7(5):.
Species: Mouse
Sample Types: Whole Cells
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Foxp3+ CD4+ regulatory T cells control dendritic cells in inducing antigen-specific immunity to emerging SARS-CoV-2 antigens
Authors: R Uraki, M Imai, M Ito, H Shime, M Odanaka, M Okuda, Y Kawaoka, S Yamazaki
PloS Pathogens, 2021-12-09;17(12):e1010085.
Species: Mouse
Sample Types: Whole Cells
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A two-adjuvant multiantigen candidate vaccine induces superior protective immune responses against SARS-CoV-2 challenge
Authors: W Jiang, L Shi, L Cai, X Wang, J Li, H Li, J Liang, Q Gu, G Ji, J Li, L Liu, M Sun
Cell Reports, 2021-11-24;37(11):110112.
Species: Mouse
Sample Types: Whole Cells
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Adaptive Immune Response to Vaccinia Virus LIVP Infection of BALB/c Mice and Protection against Lethal Reinfection with Cowpox Virus
Authors: SN Shchelkuno, AA Sergeev, SN Yakubitski, KA Titova, SA Pyankov, IV Kolosova, EV Starostina, MB Borgoyakov, AM Zadorozhny, DN Kisakov, IS Shulgina, LI Karpenko
Viruses, 2021-08-17;13(8):.
Species: Mouse
Sample Types: Whole Cells
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Combination of Photodynamic Therapy and a Flagellin-Adjuvanted Cancer Vaccine Potentiated the Anti-PD-1-Mediated Melanoma Suppression
Authors: HS Hwang, K Cherukula, YJ Bang, V Vijayan, MJ Moon, J Thiruppath, S Puth, YY Jeong, IK Park, SE Lee, JH Rhee
Cells, 2020-11-07;9(11):.
Species: Mouse
Sample Types: Whole Cells
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Synthetic mycobacterial molecular patterns partially complete Freund's adjuvant
Authors: JY Dubé, F McIntosh, JG Zarruk, S David, J Nigou, MA Behr
Sci Rep, 2020-04-03;10(1):5874.
Species: Mouse
Sample Types: Whole Cells
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Immunological Analysis of a CCHFV mRNA Vaccine Candidate in Mouse Models
Authors: T Aligholipo, K Földes, K Ergünay, H Gurdal, A Bastug, A Ozkul
Vaccines (Basel), 2019-09-16;7(3):.
Species: Mouse
Sample Types: Whole Cells
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Broad and Effective Protection against Staphylococcus aureus Is Elicited by a Multivalent Vaccine Formulated with Novel Antigens
Authors: J Deng, X Wang, BZ Zhang, P Gao, Q Lin, RY Kao, K Gustafsson, KY Yuen, JD Huang
mSphere, 2019-09-04;4(5):.
Species: Mouse
Sample Types: Whole Cells
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Extracellular overproduction of E7 oncoprotein of Iranian human papillomavirus type 16 by genetically engineered Lactococcus lactis
Authors: AH Mohseni, S Taghinezha, H Keyvani, V Razavilar
BMC Biotechnol., 2019-01-24;19(1):8.
Species: Mouse
Sample Types: Whole Cells
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Memory CD4+ T cells enhance B cell responses to drifting influenza immunization
Authors: E Gage, N Van Hoeven, ND Cauwelaert, SE Larsen, J Erasmus, MT Orr, RN Coler
Eur. J. Immunol., 2018-12-21;0(0):.
Species: Mouse
Sample Types: Whole Cells
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Protection against human papillomavirus type 16-induced tumors in C57BL/6 mice by mucosal vaccination with Lactococcus lactis NZ9000 expressing E6 oncoprotein
Authors: S Taghinezha, AH Mohseni, H Keyvani, V Razavilar
Microb. Pathog., 2018-11-02;0(0):.
Species: Mouse
Sample Types: Whole Cells
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Live attenuated Salmonella typhimurium vaccines delivering SaEsxA and SaEsxB via type III secretion system confer protection against Staphylococcus aureus infection
Authors: C Xu, BZ Zhang, Q Lin, J Deng, B Yu, S Arya, KY Yuen, JD Huang
BMC Infect. Dis., 2018-04-25;18(1):195.
Species: Mouse
Sample Types: Whole Cells
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A modified HLA-A*0201-restricted CTL epitope from human oncoprotein (hPEBP4) induces more efficient antitumor responses
Authors: W Sun, J Shi, J Wu, J Zhang, H Chen, Y Li, S Liu, Y Wu, Z Tian, X Cao, N Li
Cell. Mol. Immunol., 2018-01-29;0(0):.
Species: Mouse
Sample Types: Whole Cells
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Immunogenicity and Protective Efficacy of the DAR-901 Booster Vaccine in a Murine Model of Tuberculosis
PLoS ONE, 2016-12-20;11(12):e0168521.
Species: Mouse
Sample Types: Whole Cells
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Transient Expression of Transgenic IL-12 in Mouse Liver Triggers Unremitting Inflammation Mimicking Human Autoimmune Hepatitis
J Immunol, 2016-08-10;0(0):.
Species: Mouse
Sample Types: Whole Cells
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A whole recombinant yeast-based therapeutic vaccine elicits HBV X, S and Core specific T cells in mice and activates human T cells recognizing epitopes linked to viral clearance.
Authors: King T, Kemmler C, Guo Z, Mann D, Lu Y, Coeshott C, Gehring A, Bertoletti A, Ho Z, Delaney W, Gaggar A, Subramanian G, McHutchison J, Shrivastava S, Lee Y, Kottilil S, Bellgrau D, Rodell T, Apelian D
PLoS ONE, 2014-07-22;9(7):e101904.
Species: Mouse
Sample Types: Whole Cells
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Elongated TCR alpha chain CDR3 favors an altered CD4 cytokine profile.
Authors: Reynolds C, Chong D, Raynsford E, Quigley K, Kelly D, Llewellyn-Hughes J, Altmann D, Boyton R
BMC Biol, 2014-05-09;12(0):32.
Species: Mouse
Sample Types: Cell Culture Supernates
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Epidermal RelA specifically restricts contact allergen-induced inflammation and apoptosis in skin.
Authors: Kumari S, Herzberg B, Pofahl R, Krieg T, Haase I
J Invest Dermatol, 2014-04-16;134(10):2541-50.
Species: Mouse
Sample Types: Whole Cells
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HIV-1 adenoviral vector vaccines expressing multi-trimeric BAFF and 4-1BBL enhance T cell mediated anti-viral immunity.
Authors: Kanagavelu, Saravana, Termini, James M, Gupta, Sachin, Raffa, Francesc, Fuller, Katherin, Rivas, Yaelis, Philip, Sakhi, Kornbluth, Richard, Stone, Geoffrey
PLoS ONE, 2014-02-28;9(2):e90100.
Species: Mouse
Sample Types: Whole Cells
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Production and immunogenicity of chimeric virus-like particles containing the spike glycoprotein of infectious bronchitis virus.
Authors: Lv L, Li X, Liu G, Li R, Liu Q, Shen H, Wang W, Xue C, Cao Y
J Vet Sci, 2013-12-27;15(2):209-16.
Species: Mouse
Sample Types: Whole Cells
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Vaccination with a fusion protein that introduces HIV-1 gag antigen into a multitrimer CD40L construct results in enhanced CD8+ T cell responses and protection from viral challenge by vaccinia-gag.
Authors: Gupta, Sachin, Termini, James M, Raffa, Francesc, Williams, Cindi-An, Kornbluth, Richard, Stone, Geoffrey
J Virol, 2013-11-13;88(3):1492-501.
Species: Mouse
Sample Types: Whole Cells
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Val-boroPro accelerates T cell priming via modulation of dendritic cell trafficking resulting in complete regression of established murine tumors.
Authors: Walsh, Meghaan, Duncan, Brynn, Larabee, Shannon, Krauss, Aviva, Davis, Jessica, Cui, Yongzhi, Kim, Su Young, Guimond, Martin, Bachovchin, William, Fry, Terry J
PLoS ONE, 2013-03-12;8(3):e58860.
Species: Mouse
Sample Types: Tissue Homogenates
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The Shc family protein adaptor, Rai, acts as a negative regulator of Th17 and Th1 cell development.
Authors: Savino, Maria Te, Ulivieri, Cristina, Emmi, Giacomo, Prisco, Domenico, De Falco, Giulia, Ortensi, Barbara, Beccastrini, Enrico, Emmi, Lorenzo, Pelicci, Giuliana, D'Elios, Mario M, Baldari, Cosima T
J Leukoc Biol, 2013-01-23;93(4):549-59.
Species: Mouse
Sample Types: Whole Cells
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A VLP vaccine induces broad-spectrum cross-protective antibody immunity against H5N1 and H1N1 subtypes of influenza A virus.
Authors: Wu, Chia-Yin, Yeh, Yi-Chun, Chan, Jia-Tsro, Yang, Yu-Chih, Yang, Ji-Rong, Liu, Ming-Tsa, Wu, Ho-Sheng, Hsiao, Pei-Wen
PLoS ONE, 2012-08-07;7(8):e42363.
Species: Mouse
Sample Types: Whole Cells
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Elicitation of broad CTL response against HIV-1 by the DNA vaccine encoding artificial multi-component fusion protein MultiHIV--study in domestic pigs.
Authors: Molder T, Adojaan M, Kaldma K, Ustav M, Sikut R
Vaccine, 2009-10-29;28(2):293-8.
Species: Mouse
Sample Types: Whole Cells
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Novel immunocompetent murine tumor model for evaluation of conditionally replication-competent (oncolytic) murine adenoviral vectors.
Authors: Robinson M, Li B, Ge Y, Ko D, Yendluri S, Harding T, VanRoey M, Spindler KR, Jooss K
J. Virol., 2009-02-04;83(8):3450-62.
Species: Mouse
Sample Types: Whole Cells
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NOD2-deficient mice have impaired resistance to Mycobacterium tuberculosis infection through defective innate and adaptive immunity.
Authors: Divangahi M, Mostowy S, Coulombe F, Kozak R, Guillot L, Veyrier F, Kobayashi KS, Flavell RA, Gros P, Behr MA
J. Immunol., 2008-11-15;181(10):7157-65.
Species: Mouse
Sample Types: Whole Cells
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Type I IFNs play a role in early resistance, but subsequent susceptibility, to the African trypanosomes.
Authors: Lopez R, Demick KP, Mansfield JM, Paulnock DM
J. Immunol., 2008-10-01;181(7):4908-17.
Species: Mouse
Sample Types: Whole Cells
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Critical role of donor tissue expression of programmed death ligand-1 in regulating cardiac allograft rejection and vasculopathy.
Authors: Yang J, Popoola J, Khandwala S, Vadivel N, Vanguri V, Yuan X, Dada S, Guleria I, Tian C, Ansari MJ, Shin T, Yagita H, Azuma M, Sayegh MH, Chandraker A
Circulation, 2008-01-22;117(5):660-9.
Species: Mouse
Sample Types: Whole Cells
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Immunization with adenovirus at the large intestinal mucosa as an effective vaccination strategy against sexually transmitted viral infection.
Authors: Zhu Q, Thomson CW, Rosenthal KL, McDermott MR, Collins SM, Gauldie J
Mucosal Immunol, 2008-01-01;1(1):78-88.
Species: Mouse
Sample Types: Whole Cells
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Nitric oxide mediates T cell cytokine production and signal transduction in histidine decarboxylase knockout mice.
Authors: Koncz A, Pasztoi M, Mazan M, Fazakas F, Buzas E, Falus A, Nagy G
J. Immunol., 2007-11-15;179(10):6613-9.
Species: Mouse
Sample Types: Whole Cells
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One-year Expression From High-capacity Adenoviral Vectors in the Brains of Animals With Pre-existing Anti-adenoviral Immunity: Clinical Implications.
Authors: Barcia C, Jimenez-Dalmaroni M, Kroeger KM, Puntel M, Rapaport AJ, Larocque D, King GD, Johnson SA, Liu C, Xiong W, Candolfi M, Mondkar S, Ng P, Palmer D, Castro MG, Lowenstein PR
Mol. Ther., 2007-09-25;15(12):2154-2163.
Species: Mouse
Sample Types: Whole Cells
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Inactivated Sendai virus particles eradicate tumors by inducing immune responses through blocking regulatory T cells.
Authors: Kurooka M, Kaneda Y
Cancer Res., 2007-01-01;67(1):227-36.
Species: Mouse
Sample Types: Whole Cells
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Elicitation of immunity to HIV type 1 Gag is determined by Gag structure.
Authors: Young KR, Ross TM
AIDS Res. Hum. Retroviruses, 2006-01-01;22(1):99-108.
Species: Mouse
Sample Types: Whole Cells
Applications: ELISpot Development -
Immunopharmacological and antitumor effects of second-generation immunomodulatory oligonucleotides containing synthetic CpR motifs.
Authors: Wang D, Li Y, Yu D, Song SS, Kandimalla ER, Agrawal S
Int. J. Oncol., 2004-04-01;24(4):901-8.
Species: Mouse
Sample Types: Whole Cells
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