IFN-gamma Antibody (37895) [Unconjugated]

Mouse splenocytes either (A) stimulated to induce Th1 cells or (B) unstimulated were stained with Rat Anti-Mouse IFN-gamma Monoclonal Antibody (Catalog # MAB485) followed by Phycoerythrin-conjugated Anti-Rat IgG Secondary Antibody (F0105B) and Rat Anti-Mouse CD4 APC-conjugated Monoclonal Antibody (FAB554A). Quadrant markers were set based on control antibody staining (MAB006). To facilitate intracellular staining, cells were fixed with Flow Cytometry Fixation Buffer (FC004) and permeabilized with Flow Cytometry Permeabilization/Wash Buffer I (FC005). View our protocol for Staining Intracellular Molecules.

Mouse splenocytes either (A) stimulated for 4 hours with Cell Activation Cocktail 500x (5476) or (B) unstimulated were stained with Rat Anti-Mouse IFN-gamma Monoclonal Antibody (Catalog # MAB485) followed by Phycoerythrin-conjugated Anti-Rat IgG Secondary Antibody (F0105B) and Rat Anti-Mouse CD4 APC-conjugated Monoclonal Antibody (FAB554A). Quadrant markers were set based on control antibody staining (MAB006). To facilitate intracellular staining, cells were fixed with Flow Cytometry Fixation Buffer (FC004) and permeabilized with Flow Cytometry Permeabilization/Wash Buffer I (FC005). Staining was performed using our Staining Intracellular Molecules protocol.

Recombinant Mouse IFN-gamma 485-MI) reduces the Encephalomyocarditis Virus (EMCV)-induced cytopathy in the L-929 mouse fibroblast cell line in a dose-dependent manner (orange line), as measured by crystal violet staining. Inhibition of EMCV activity elicited by Recombinant Mouse IFN-gamma (2.5 ng/mL) is neutralized (green line) by increasing concentrations of Rat Anti-Mouse IFN-gamma Monoclonal Antibody (Catalog # MAB485). The ND50 is typically 0.075-0.3 µg/mL.

IFN‑ gamma was detected in immersion fixed paraffin-embedded sections of Mouse Spleen using Rat Anti-Mouse IFN‑ gamma Monoclonal Antibody (Catalog # MAB485) at 10 µg/mL for 1 hour at room temperature followed by incubation with the Anti-Rat IgG VisUCyte™ HRP Polymer Antibody (Catalog # VC005). Before incubation with the primary antibody, tissue was subjected to heat-induced epitope retrieval using VisUCyte Antigen Retrieval Reagent-Basic (Catalog # VCTS021). Tissue was stained using DAB (brown) and counterstained with hematoxylin (blue). Specific staining was localized to cytoplasm in lymphocytes. View our protocol for IHC Staining with VisUCyte HRP Polymer Detection Reagents.

EAE induction in KO mice.(a) Means±s.e.m. of EAE clinical scores of KO and WT mice. *P<0.05 (two-tailed Student's t-test). (b) EAE incidence in KO and WT mice. *P<0.05 (chi-square test). (c) Means±s.e.m. of body weight of KO and WT mice during EAE induction. Body weight of mice on day 10 post-immunization was considered as 100%. *P<0.05 (two-tailed Student's t-test). (d) Means±s.e.m. of cellularity in draining LN and of cells infiltrating the CNS of mice 14 days after MOG immunization. Mouse numbers (n) and P values (paired two-tailed Student's t-test) are indicated. (e,f) Cytokine-producing cells among CD4 cells from draining LN (e) and CNS (f) on days 13–18 after MOG immunization. Left panels: representative dot plots; right panel: bar graphs (means±s.e.m.) summarizing all the results, with mouse numbers and P values (two-tailed Student's t-test) indicated. (g) HE (left column) or Luxol Fast Blue (right column) staining of spinal cords 30 days after MOG immunization. Asterisks indicate cell infiltration. Arrows point to demyelination. (h) Means±s.e.m. of mononuclear cell infiltration scores, demyelination scores and total pathological scores, which is the sum of the first two scores. Mouse numbers (n) and P values (two-tailed Student's t-test) are indicated. (i) Treg cells in naive KO mice on day 17 during EAE induction. Left panel: representative dot plots; right panel: means±s.e.m. of data from three experiments. NS: not significant (two-tailed Student's t-test). Image collected and cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/28169274), licensed under a CC-BY license. Not internally tested by R&D Systems.

EAE induction in KO mice.(a) Means±s.e.m. of EAE clinical scores of KO and WT mice. *P<0.05 (two-tailed Student's t-test). (b) EAE incidence in KO and WT mice. *P<0.05 (chi-square test). (c) Means±s.e.m. of body weight of KO and WT mice during EAE induction. Body weight of mice on day 10 post-immunization was considered as 100%. *P<0.05 (two-tailed Student's t-test). (d) Means±s.e.m. of cellularity in draining LN and of cells infiltrating the CNS of mice 14 days after MOG immunization. Mouse numbers (n) and P values (paired two-tailed Student's t-test) are indicated. (e,f) Cytokine-producing cells among CD4 cells from draining LN (e) and CNS (f) on days 13–18 after MOG immunization. Left panels: representative dot plots; right panel: bar graphs (means±s.e.m.) summarizing all the results, with mouse numbers and P values (two-tailed Student's t-test) indicated. (g) HE (left column) or Luxol Fast Blue (right column) staining of spinal cords 30 days after MOG immunization. Asterisks indicate cell infiltration. Arrows point to demyelination. (h) Means±s.e.m. of mononuclear cell infiltration scores, demyelination scores and total pathological scores, which is the sum of the first two scores. Mouse numbers (n) and P values (two-tailed Student's t-test) are indicated. (i) Treg cells in naive KO mice on day 17 during EAE induction. Left panel: representative dot plots; right panel: means±s.e.m. of data from three experiments. NS: not significant (two-tailed Student's t-test). Image collected and cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/28169274), licensed under a CC-BY license. Not internally tested by R&D Systems.

Proliferation and differentiation of naive KO CD4 cells into Th1 and Th17 cells.(a) Proliferation of WT and KO naive spleen CD4 cells under Th1 and Th17 conditions was assessed based on CFSE content according to flow cytometry. Experiments were conducted three times, and representative histograms are shown. Grey peaks represent the CFSE content of CD4 cells at day 0. (b) These cells' differentiation into Th1 and Th17 cells was also determined by flow cytometry according to intracellular IFN-gamma and IL-17 positivity (gated on total CD4+). Representative dot plots are shown in the left panel. Means±s.e.m. of data from three experiments are presented as bar graphs in the right panel. Mouse numbers (n) per group are indicated. P values are reported in the bar graphs (two-tailed Student's t-test). (c,d) T-bet and ROR gamma t expression in CD4 cells cultured under Th1 and Th17 conditions or in IFN gamma + or IL-17+ cells was determined by flow cytometry. Experiments were conducted three times. Representative histograms are shown. (e) Th1 and Th17 differentiation of naive spleen CD4 cells (CD45.2 single-positive) derived from WT and KO donors in chimeric mice was analysed by flow cytometry based on their intracellular IFN-gamma and IL-17 expression. Representative dot plots are shown in the left panel. Means±s.e.m. of data from three experiments are presented as bar graphs in the right panel. Mouse numbers (n) per group are indicated. p values are reported in the bar graphs (two-tailed Student's t-test). Image collected and cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/28169274), licensed under a CC-BY license. Not internally tested by R&D Systems.

Proliferation and differentiation of naive KO CD4 cells into Th1 and Th17 cells.(a) Proliferation of WT and KO naive spleen CD4 cells under Th1 and Th17 conditions was assessed based on CFSE content according to flow cytometry. Experiments were conducted three times, and representative histograms are shown. Grey peaks represent the CFSE content of CD4 cells at day 0. (b) These cells' differentiation into Th1 and Th17 cells was also determined by flow cytometry according to intracellular IFN-gamma and IL-17 positivity (gated on total CD4+). Representative dot plots are shown in the left panel. Means±s.e.m. of data from three experiments are presented as bar graphs in the right panel. Mouse numbers (n) per group are indicated. P values are reported in the bar graphs (two-tailed Student's t-test). (c,d) T-bet and ROR gamma t expression in CD4 cells cultured under Th1 and Th17 conditions or in IFN gamma + or IL-17+ cells was determined by flow cytometry. Experiments were conducted three times. Representative histograms are shown. (e) Th1 and Th17 differentiation of naive spleen CD4 cells (CD45.2 single-positive) derived from WT and KO donors in chimeric mice was analysed by flow cytometry based on their intracellular IFN-gamma and IL-17 expression. Representative dot plots are shown in the left panel. Means±s.e.m. of data from three experiments are presented as bar graphs in the right panel. Mouse numbers (n) per group are indicated. p values are reported in the bar graphs (two-tailed Student's t-test). Image collected and cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/28169274), licensed under a CC-BY license. Not internally tested by R&D Systems.

• Reactivity: Mu
• Applications: WB, IHC, B/N, CyTOF-ready, Flow
• Clone: 37895
• Clonality: Monoclonal
• Host: Rat
• Conjugate: Unconjugated

IFN-gamma Antibody (37895) [Unconjugated] Summary

• Immunogen: E. coli-derived recombinant mouse IFN-gamma
• Specificity: Detects mouse IFN-gamma in Western blots. In Western blots, this antibody does not cross-react with recombinant human (rh) IFN‑ gamma , rrIFN‑ gamma , rpIFN‑ gamma , rrmIFN‑ gamma , rfeIFN‑ gamma , or rcaIFN‑ gamma .
• Source: N/A
• Isotype: IgG2a
• Clonality: Monoclonal
• Host: Rat
• Gene: IFNG
• Purity Statement: Protein A or G purified from hybridoma culture supernatant
• Endotoxin Note: <0.10 EU per 1 μg of the antibody by the LAL method.

Applications/Dilutions

• Dilutions:
  • CyTOF-ready
  • Immunohistochemistry 5-15 ug/mL
  • Intracellular Staining by Flow Cytometry 0.25 ug/10^6 cells
  • Neutralization 0.075-0.3 ug/mL
  • Western Blot 1 ug/mL

Packaging, Storage & Formulations

• Storage: Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
  • 12 months from date of receipt, -20 to -70 °C as supplied.
  • 1 month, 2 to 8 °C under sterile conditions after reconstitution.
  • 6 months, -20 to -70 °C under sterile conditions after reconstitution.
• Buffer: Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied either lyophilized or as a 0.2 µm filtered solution in PBS.
• Preservative: No Preservative
• Reconstitution Instructions: Reconstitute at 0.5 mg/mL in sterile PBS.

Notes

This product is produced by and ships from R&D Systems, Inc., a Bio-Techne brand.

Alternate Names for IFN-gamma Antibody (37895) [Unconjugated]

• IFG
• IFI
• IFNG
• IFNgamma
• IFN-gamma
• Immune interferon
• interferon gamma
• interferon, gamma

Background

Interferon-gamma (IFN-gamma, IFNG), also known as type II or Immune Interferon, exerts a wide range of immunoregulatory activities and is considered to be the prototype proinflammatory cytokine. Mature human IFN-gamma exists as a non-covalently linked homodimer of 20-25 kDa molecular weight variably glycosylated subunits. It shares 86% amino acid (aa) sequence identity with rat IFN-gamma, 38-44% with bovine, canine, cotton rat, equine, feline, human, porcine and rhesus IFN‑ gamma. IFN-gamma dimers bind to IFN-gamma RI (alpha subunits) which then interact with IFN-gamma RII (beta subunits) to form the functional receptor complex of two alpha and two beta subunits. Inclusion of IFN-gamma RII increases the binding affinity for ligand and the efficiency of signal transduction. IFN-gamma is produced by a variety of immune cells under inflammatory conditions, notably by T cells and NK cells. It plays a key function in host defense by promoting the development and activation of Th1 cells, chemoattraction and activation of monocytes and macrophages, up‑regulation 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. The pleiotropic effects of IFN-gamma contribute to the development of multiple aspects of atherosclerosis.

Limitations

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

Publications for IFN-gamma Antibody (MAB485)(32)

Publications using MAB485

• Merchak, AR;Wachamo, S;Brown, LC;Thakur, A;Moreau, B;Brown, RM;Rivet-Noor, C;Raghavan, T;Gaultier, A; Lactobacillus from the Altered Schaedler Flora maintain IFN? homeostasis to promote behavioral stress resilience Brain, behavior, and immunity 2023-11-02 [PMID: 37924959] (In Vivo, Mouse)
• JS Park, SC Yang, HY Jeong, SY Lee, JG Ryu, JW Choi, HY Kang, SM Kim, SH Hwang, ML Cho, SH Park EC-18 prevents autoimmune arthritis by suppressing inflammatory cytokines and osteoclastogenesis Arthritis Research & Therapy, 2022-11-17;24(1):254. 2022-11-17 [PMID: 36397156] (Neutralization, Mouse)
• T Zhao, S Liu, X Ding, EM Johnson, NH Hanna, K Singh, CK Sen, J Wan, H Du, C Yan Lysosomal acid lipase, CSF1R and PD-L1 determine functions of CD11c+ myeloid-derived suppressor cells JCI Insight, 2022-09-08;0(0):. 2022-09-08 [PMID: 35917184] (Neutralization, Mouse)
• RJ Napier, EJ Lee, MP Davey, EE Vance, JM Furtado, PE Snow, KA Samson, SJ Lashley, BR Brown, R Horai, MJ Mattapalli, B Xu, MC Callegan, LS Uebelhoer, CL Lancioni, RK Vehe, BA Binstadt, JR Smith, RR Caspi, HL Rosenzweig T cell-intrinsic role for Nod2 in protection against Th17-mediated uveitis Nat Commun, 2020-10-26;11(1):5406. 2020-10-26 [PMID: 33106495] (Neutralization, Mouse)
• JQ Yang, KW Kalim, Y Li, Y Zheng, F Guo Ablation of RhoA impairs Th17 cell differentiation and alleviates house dust mite-triggered allergic airway inflammation J. Leukoc. Biol., 2019-07-01;0(0):. 2019-07-01 [PMID: 31260596] (Bioassay, Mouse)
• Laura Evgin, Amanda L. Huff, Timothy Kottke, Jill Thompson, Amy M. Molan, Christopher B. Driscoll et al. Suboptimal T-cell Therapy Drives a Tumor Cell Mutator Phenotype That Promotes Escape from First-Line Treatment Cancer Immunology Research 2019-05-01 [PMID: 30940643]
• Su‑Ping Xu, Yong‑Sheng Li Fisetin inhibits pristine-induced systemic lupus erythematosus in a murine model through CXCLs regulation International Journal of Molecular Medicine 2018-10-01 [PMID: 30272314]
• Y Nam, SM Jung, YA Rim, H Jung, K Lee, N Park, J Kim, Y Jang, YB Park, SH Park, JH Ju Intraperitoneal infusion of mesenchymal stem cell attenuates severity of collagen antibody induced arthritis PLoS ONE, 2018-06-07;13(6):e0198740. 2018-06-07 [PMID: 29879214] (Functional Assay, Mouse)
• W Lei, D Zeng, G Liu, Y Zhu, J Wang, H Wu, J Jiang, J Huang Crucial role of OX40/OX40L signaling in a murine model of asthma Mol Med Rep, 2018-01-18;0(0):. 2018-01-18 [PMID: 29344664] (Functional Assay, Mouse)
• Wei Cao, Hisako Kayama, Mei Lan Chen, Amber Delmas, Amy Sun, Sang Yong Kim et al. The Xenobiotic Transporter Mdr1 Enforces T Cell Homeostasis in the Presence of Intestinal Bile Acids Immunity 2017-12-01 [PMID: 29262351]
• Pawan Kumar, Jay K Kolls Lymphocyte Isolation, Th17 Cell Differentiation, Activation, and Staining BIO-PROTOCOL 2016-12-05 [PMID: 29104893]
• Y Hu, L Lao, J Mao, W Jin, H Luo, T Charpentie, S Qi, J Peng, B Hu, MM Marcinkiew, A Lamarre, J Wu Armc5 deletion causes developmental defects and compromises T-cell immune responses Nat Commun, 2017-02-07;8(0):13834. 2017-02-07 [PMID: 28169274] (Neutralization, Mouse)
• Jonathan M. Weiss, Wei Chen, Melanie S. Nyuydzefe, Alissa Trzeciak, Ryan Flynn, James R. Tonra et al. ROCK2 signaling is required to induce a subset of T follicular helper cells through opposing effects on STATs in autoimmune settings Science Signaling 7/21/2016 [PMID: 27436361]
• Ivan D. Mascanfroni, Maisa C. Takenaka, Ada Yeste, Bonny Patel, Yan Wu, Jessica E. Kenison et al. Metabolic control of type 1 regulatory (Tr1) cell differentiation by AHR and HIF1-alpha Nature Medicine 2015-06-01 [PMID: 26005855]
• Lee H, Ko J, Jeong H, Ko A, Kim M, Wee W, Yoon S, Oh J Mesenchymal stem/stromal cells protect against autoimmunity via CCL2-dependent recruitment of myeloid-derived suppressor cells. J Immunol, 2015-03-13;194(8):3634-45. 2015-03-13 [PMID: 25769927] (Bioassay, Mouse)
• Wunnie Brima, Daniel J. Eden, Syed Faizan Mehdi, Michelle Bravo, Mohammad M. Wiese, Joanna Stein et al. The brighter (and evolutionarily older) face of the metabolic syndrome: evidence from Trypanosoma cruzi infection in CD-1 mice Diabetes/Metabolism Research and Reviews 2015-05-01 [PMID: 25613819]
• He W, Yang C, Xia L, Zhao M, Ge R, Huang H, Ji Q, Yang P, Liu Z CD4(+) T cells from food allergy model are resistant to TCR-dependent apoptotic induction. Cytokine, 2014-04-16;68(1):32-9. 2014-04-16 [PMID: 24787054] (Neutralization, Mouse)
• Yoo , Hyeon-Se, Choi , Yongwook, Ahn , Narae, Lee , Saseong, Kim , Wan-Uk, Jang , Min Seon, Jang , Myoung H, Kim , Yon Su, Yoo , Joo-Yeon Transcriptional regulator CTR9 inhibits Th17 differentiation via repression of IL-17 expression. J Immunol, 2014-01-13;192(4):1440-8. 2014-01-13 [PMID: 24420920] (Functional Assay, Mouse)
• Francisco J Quintana, Hulin Jin, Evan J Burns, Meghan Nadeau, Ada Yeste, Deepak Kumar et al. Aiolos promotes TH17 differentiation by directly silencing Il2 expression Nature Immunology 2012-08-01 [PMID: 22751139]
• Jin W, Qi S, Luo H The effect of conditional EFNB1 deletion in the T cell compartment on T cell development and function. BMC Immunol., 2011-12-19;12(0):68. 2011-12-19 [PMID: 22182253] (Neutralization, Mouse)
• Mizuho Nosaka, Yuko Ishida, Akihiko Kimura, Yumi Kuninaka, Masanori Inui, Naofumi Mukaida et al. Absence of IFN-gamma accelerates thrombus resolution through enhanced MMP-9 and VEGF expression in mice Journal of Clinical Investigation 2011-07-01 [PMID: 21646723]
• Sarra M, Caruso R, Cupi ML, Monteleone I, Stolfi C, Campione E, Diluvio L, Mazzotta A, Botti E, Chimenti S, Costanzo A, MacDonald TT, Pallone F, Monteleone G IL-21 Promotes Skin Recruitment of CD4+ Cells and Drives IFN-{gamma}-Dependent Epidermal Hyperplasia. J. Immunol., 2011-03-25;186(9):5435-42. 2011-03-25 [PMID: 21441456] (Neutralization, Mouse)
• Jones GW, Stumhofer JS, Foster T, Twohig JP, Hertzog P, Topley N, Williams AS, Hunter CA, Jenkins BJ, Wang EC, Jones SA Naive and activated T cells display differential responsiveness to TL1A that affects Th17 generation, maintenance, and proliferation. FASEB J., 2010-09-08;25(1):409-19. 2010-09-08 [PMID: 20826539] (Neutralization, Mouse)
• Horvat JC, Starkey MR, Kim RY, Beagley KW, Preston JA, Gibson PG, Foster PS, Hansbro PM Chlamydial respiratory infection during allergen sensitization drives neutrophilic allergic airways disease. J. Immunol., 2010-03-12;184(8):4159-69. 2010-03-12 [PMID: 20228193] (Neutralization, Mouse)
• Deepak P, Kumar S, Kishore D, Acharya A IL-13 from Th2-type cells suppresses induction of antigen-specific Th1 immunity in a T-cell lymphoma. Int. Immunol., 2009-11-30;22(1):53-63. 2009-11-30 [PMID: 19951958] (ELISA Development, Mouse)
• McKinley L, Alcorn JF, Peterson A, Dupont RB, Kapadia S, Logar A, Henry A, Irvin CG, Piganelli JD, Ray A, Kolls JK TH17 cells mediate steroid-resistant airway inflammation and airway hyperresponsiveness in mice. J. Immunol., 2008-09-15;181(6):4089-97. 2008-09-15 [PMID: 18768865] (Neutralization, Mouse)
• Mabalirajan U, Dinda AK, Kumar S, Roshan R, Gupta P, Sharma SK, Ghosh B Mitochondrial structural changes and dysfunction are associated with experimental allergic asthma. J. Immunol., 2008-09-01;181(5):3540-8. 2008-09-01 [PMID: 18714027] (Neutralization, Mouse)
• Iwanami K, Matsumoto I, Tanaka-Watanabe Y, Inoue A, Mihara M, Ohsugi Y, Mamura M, Goto D, Ito S, Tsutsumi A, Kishimoto T, Sumida T Crucial role of the interleukin-6/interleukin-17 cytokine axis in the induction of arthritis by glucose-6-phosphate isomerase. Arthritis Rheum., 2008-03-01;58(3):754-63. 2008-03-01 [PMID: 18311788] (Neutralization, Mouse)
• Nakamura M, Jo J, Tabata Y, Ishikawa O Controlled delivery of T-box21 small interfering RNA ameliorates autoimmune alopecia (Alopecia Areata) in a C3H/HeJ mouse model. Am. J. Pathol., 2008-02-02;172(3):650-8. 2008-02-02 [PMID: 18245811] (Neutralization, Mouse)
• Smith SE, Li J, Garbett K, Mirnics K, Patterson PH Maternal immune activation alters fetal brain development through interleukin-6. J. Neurosci., 2007-10-03;27(40):10695-702. 2007-10-03 [PMID: 17913903] (Neutralization, Mouse)
• Haraguchi G, Suzuki J, Kosuge H, Ogawa M, Koga N, Muto S, Itai A, Kagechika H, Shudo K, Isobe M A new RXR agonist, HX630, suppresses intimal hyperplasia in a mouse blood flow cessation model. J. Mol. Cell. Cardiol., 2006-09-11;41(5):885-92. 2006-09-11 [PMID: 16963076] (IHC, Mouse)
• Kamath AT, Sheasby CE, Tough DF Dendritic cells and NK cells stimulate bystander T cell activation in response to TLR agonists through secretion of IFN-alpha beta and IFN-gamma. J. Immunol., 2005-01-15;174(2):767-76. 2005-01-15 [PMID: 15634897] (Flow Cytometry, Neutralization, Mouse)

FAQs for IFN-gamma Antibody (MAB485). (Showing 1 - 1 of 1 FAQs).

1. Is Recombinant Human IFN-gamma Protein - Clinical grade or preclinical ?
   • We are research use only.

Bioinformatics

• Gene Symbol: IFNG