TNF alpha Monoclonal Antibody (MP6-XT22), FITC, ebioscience Catalog Number Product data sheet

Transcription

1 Website: thermofisher.com/ebioscience Customer Service (US): thermofisher.com/contactus TNF alpha Monoclonal Antibody (MP6-XT22), FITC, ebioscience Catalog Number Product data sheet Details Size 50 µg Host/Isotope Class Type Clone Conjugate Form Concentration Purification Storage buffer Contains Storage Conditions Rat / IgG1, kappa Monoclonal Antibody MP6-XT22 FITC Liquid 0.5 mg/ml Affinity chromatography PBS, ph 7.2, with 0.1% gelatin 0.09% sodium azide 4 C, store in dark, DO NOT FREEZE! Species Reactivity Tested species reactivity Published species reactivity Mouse Tested Applications Dilution * Flow Cytometry (Flow) Published Applications Flow Cytometry (Flow) Immunocytochemistry (ICC) ELISA (ELISA) Immunofluorescence (IF) Immunohistochemistry (Frozen) (IHC (F)) Immunohistochemistry (IHC) Miscellaneous PubMed (MISC) Functional Assay (FN) Mouse, Not Applicable µg/test See 22 publications below See 2 publications below See 2 publications below * Suggested working dilutions are given as a guide only. It is recommended that the user titrate the product for use in their own experiment using appropriate negative and positive controls. Product specific information Description: The MP6-XT22 antibody reacts with mouse tumor necrosis factor-alpha (TNF alpha), a 17 kda cytokine produced by monocytes, macrophages, neutrophils, NK cells and CD4(+)T cells. TNF alpha has cytolytic activity against a range of tumor cells and is important in immune regulation. TNF alpha forms dimers and trimers and also exists as a 26 kda membrane-bound form. Applications Reported: The MP6-XT22 antibody has been reported for use in ELISA, neutralization, and intracellular staining for flow cytometric analysis. Applications Tested: The MP6-XT22 antibody is offered in 2 formats: - µg size: has been tested by intracellular flow cytometric analysis. This can be used at less than or equal to µg per test. A test is defined as the amount (µg) of antibody that will stain a cell sample in a final volume of 100 µl. Cell number should be determined empirically but can range from 10^5 to 10^8 cells/test. It is recommended that the antibody be carefully titrated for optimal performance in the assay of interest. - test size: has been pre-titrated and tested by intracellular flow cytometric analysis. This can be used at test size: 5 µl (0.06 µg) per test. Excitation: 488 nm; Emission: 520 nm; Laser: Blue Laser. Filtration: 0.2 µm post-manufacturing filtered. Background/Target Information Tumor Necrosis Factor-Alpha (TNF alpha) is a protein secreted by lipopolysaccharide-stimulated macrophages, and causes tumor necrosis when injected into tumor-bearing mice. TNF alpha is believed to mediate pathogenic shock and tissue injury associated with endotoxemia. TNF alpha exists as a multimer of two, three, or five noncovalently linked units, but shows a single ~17 kda band following SDS-PAGE under non-reducing conditions.

2 Product Images For TNF alpha Monoclonal Antibody (MP6-XT22), FITC, ebioscience TNF alpha Antibody in Immunohistochemistry (IHC) Published figure using TNF alpha monoclonal antibody (Product # ) in Immunohistochemistry TNF alpha Antibody ( ) Mouse splenocytes were stimulated with ConA for 2 days, followed by Mouse IL-2 Recombinant Protein (Product # ) and Mouse IL-4 Recombinant Protein (Product # ) for 3 days, and restimulated with immobilized Anti-Mouse CD3 Functional Grade Purified (Product # ) and soluble Anti-Mouse CD28 Functional Grade Purified (Product # ) in the presence of Brefeldin A (Product # ) for 5 hours. Cells were stained with Anti-Mouse CD4 PE (Product # ) and 0.06 µg of Rat IgG1 K Isotype Control FITC (Product # ) (left) or 0.06 µg of Anti-Mouse TNF alpha FITC (right). Cells in the lymphocyte gate were used for analysis. TNF alpha Antibody in Immunohistochemistry (IHC) Published figure using TNF alpha monoclonal antibody (Product # ) in Immunohistochemistry

3 PubMed References For TNF alpha Monoclonal Antibody (MP6-XT22), FITC, ebioscience 22 Flow Cytometry References was used in Flow cytometry/cell sorting to determine the effect of CCR4 antagonism on human renal cell carcinoma. Mouse / 1:75 Mouse / 1:400 Mouse / 1:100 The Journal of clinical investigation (Mar 2017; 127: 801) "A CCR4 antagonist reverses the tumor-promoting microenvironment of renal cancer." Author(s):Berlato C,Khan MN,Schioppa T,Thompson R,Maniati E,Montfort A,Jangani M,Canosa M,Kulbe H,Hagemann UB, Duncan AR,Fletcher L,Wilkinson RW,Powles T,Quezada SA,Balkwill FR PubMed Article URL: was used in Flow cytometry/cell sorting to evaluate how autophagy inhibition impacts the antitumor immune response in immune-competent mouse models of melanoma and mammary cancer. The Journal of clinical investigation (Dec 2016; 126: 4417) "Antitumor adaptive immunity remains intact following inhibition of autophagy and antimalarial treatment." Author(s):Starobinets H,Ye J,Broz M,Barry K,Goldsmith J,Marsh T,Rostker F,Krummel M,Debnath J PubMed Article URL: was used in Flow cytometry/cell sorting to uncover an eomesodermin-driven CD4 T cell program that is continuously suppressed by TGF- signalling. The Journal of clinical investigation (Oct 2016; 126: 3799) "TGF- receptor maintains CD4 T helper cell identity during chronic viral infections." Author(s):Lewis GM,Wehrens EJ,Labarta-Bajo L,Streeck H,Zuniga EI PubMed Article URL: was used in Flow cytometry/cell sorting to investigate the role of of CD4(+) T-cell help in CD8(+) T-cell responses against viral infections. Cellular and molecular immunology (Jan 2016; 13: 82) "CD4(+) T-cell dependence of primary CD8(+) T-cell response against vaccinia virus depends upon route of infection and viral dose." Author(s):Hu Z,Molloy MJ,Usherwood EJ PubMed Article URL: was used in Flow cytometry/cell sorting to investigate the nature of defects of T-cell immunity in post-acute phases of systemic inflammatory response, showing an induced loss of naive T-cells but no enduring cell-autonomous defects in T- cell function. PloS one (Aug 2015; 9: null) "Immunosuppression after sepsis: systemic inflammation and sepsis induce a loss of naïve T-cells but no enduring cell-autonomous defects in T-cell function." Author(s):Markwart R,Condotta SA,Requardt RP,Borken F,Schubert K,Weigel C,Bauer M,Griffith TS,Förster M,Brunkhorst FM, Badovinac VP,Rubio I PubMed Article URL: was used in Flow cytometry/cell sorting to determine whether tumour vasculature influences the functional capacity of tumour infiltrating lymphocytes, and correlate this with survival in ovarian cancer patients. PloS one (Mar 2015; 8: null) "Markers of T cell infiltration and function associate with favorable outcome in vascularized high-grade serous ovarian carcinoma." Author(s):Townsend KN,Spowart JE,Huwait H,Eshragh S,West NR,Elrick MA,Kalloger SE,Anglesio M,Watson PH,Huntsman DG,Lum JJ PubMed Article URL: was used in Flow cytometry/cell sorting to demonstrate that the recovery of CD4 T cells from sepsis-induced lymphopenia is accompanied by alterations to the composition and function of the Ag-specific CD4 T cell repertoire. Journal of immunology (Baltimore, Md. : 1950) (Feb 2015; 194: 1609) "Alterations in antigen-specific naive CD4 T cell precursors after sepsis impairs their responsiveness to pathogen challenge." Author(s):Cabrera-Perez J,Condotta SA,James BR,Kashem SW,Brincks EL,Rai D,Kucaba TA,Badovinac VP,Griffith TS PubMed Article URL:

4 was used in Flow cytometry/cell sorting to investigate the very early host-virus interactions that determine the establishment of viral persistence. PLoS pathogens (Jan 2015; 11: null) "Early virus-host interactions dictate the course of a persistent infection." Author(s):Sullivan BM,Teijaro JR,de la Torre JC,Oldstone MB PubMed Article URL: was used in Flow cytometry/cell sorting to investigate the importance of GM-CSF in the development of severe interstitial lung disease. Journal of immunology (Baltimore, Md. : 1950) (Jul 2014; 193: 849) "GM-CSF but not IL-17 is critical for the development of severe interstitial lung disease in SKG mice." Author(s):Shiomi A,Usui T,Ishikawa Y,Shimizu M,Murakami K,Mimori T PubMed Article URL: was used in Flow cytometry/cell sorting to assess the effectiveness of different immunotherapy combination treatments against the murine B16.SIY melanoma model. Journal for immunotherapy of cancer (May 2014; 2: null) "Mechanism of tumor rejection with doublets of CTLA-4, PD-1/PD-L1, or IDO blockade involves restored IL-2 production and proliferation of CD8(+) T cells directly within the tumor microenvironment." Author(s):Spranger S,Koblish HK,Horton B,Scherle PA,Newton R,Gajewski TF PubMed Article URL: was used in Flow cytometry/cell sorting to investigate the intestinal immunopathology caused by insufficient Roquin, showing that hematopoietic, not systematic, impairment of Roquin expression accounts for pathology. Scientific reports (May 2014; 4: null) "Hematopoietic not systemic impairment of Roquin expression accounts for intestinal inflammation in Roquindeficient mice." Author(s):Montufar-Solis D,Vigneswaran N,Nakra N,Schaefer JS,Klein JR PubMed Article URL: was used in Flow cytometry/cell sorting to study an rsfv-based vector which induces robust T-cell activity and has a potential for the treatment of HCV infections. Molecular therapy : the journal of the American Society of Gene Therapy (Apr 2014; 22: 881) "Alphavirus-based vaccines encoding nonstructural proteins of hepatitis C virus induce robust and protective T-cell responses." Author(s):Ip PP,Boerma A,Regts J,Meijerhof T,Wilschut J,Nijman HW,Daemen T PubMed Article URL: was used in Flow cytometry/cell sorting to investigate the efficacy of cytotoxic T cell induction by a novel polyvalentfilovirus DNA vaccine against Marburg and Ebola. Molecular therapy : the journal of the American Society of Gene Therapy (Jul 2013; 21: 1432) "Induction of broad cytotoxic T cells by protective DNA vaccination against Marburg and Ebola." Author(s):Shedlock DJ,Aviles J,Talbott KT,Wong G,Wu SJ,Villarreal DO,Myles DJ,Croyle MA,Yan J,Kobinger GP,Weiner DB PubMed Article URL: was used in Flow cytometry/cell sorting to assess transforming growth factor beta inhibition as a viable approach to control a persistent viral infection. Journal of virology (Jul 2012; 86: 7060) "Therapeutic blockade of transforming growth factor beta fails to promote clearance of a persistent viral infection." Author(s):Garidou L,Heydari S,Gossa S,McGavern DB PubMed Article URL: was used in Flow cytometry/cell sorting to investigate the use of Parapoxvirus ovis for the treatment of cancer. Molecular therapy : the journal of the American Society of Gene Therapy (Jun 2012; 20: 1148) "ORFV: a novel oncolytic and immune stimulating parapoxvirus therapeutic." Author(s):Rintoul JL,Lemay CG,Tai LH,Stanford MM,Falls TJ,de Souza CT,Bridle BW,Daneshmand M,Ohashi PS,Wan Y, Lichty BD,Mercer AA,Auer RC,Atkins HL,Bell JC PubMed Article URL:

5 was used in Flow cytometry/cell sorting to determine the usefulness of a CBA/J-ML29 mouse model as a tool for evaluating the efficacy of vaccine candidates against LASV. Mouse / 1:100 Vaccine (Feb 2012; 30: 1445) "Evaluation of Lassa virus vaccine immunogenicity in a CBA/J-ML29 mouse model." Author(s):Goicochea MA,Zapata JC,Bryant J,Davis H,Salvato MS,Lukashevich IS PubMed Article URL: was used in Flow cytometry/cell sorting to study changes in expression of Tbet by CD4 T cells from distinct areas of the body following the onset of experimental autoimmune encephalomyelitis. Journal of immunology (Baltimore, Md. : 1950) (Nov 2011; 187: 4998) "Autoreactive Tbet-positive CD4 T cells develop independent of classic Th1 cytokine signaling during experimental autoimmune encephalomyelitis." Author(s):Yeh WI,McWilliams IL,Harrington LE PubMed Article URL: was used in Flow cytometry/cell sorting to study how TRAIL affects the T cell response and ultimately influences the outcome of viral infection. Journal of immunology (Baltimore, Md. : 1950) (Nov 2011; 187: 4581) "The magnitude of the T cell response to a clinically significant dose of influenza virus is regulated by TRAIL." Author(s):Brincks EL,Gurung P,Langlois RA,Hemann EA,Legge KL,Griffith TS PubMed Article URL: was used in Flow cytometry/cell sorting to study how the preferential replication of VACV in the ovaries may not be dependent on immune suppressive mechanisms in this tissue. Viral immunology (Oct 2011; 24: 387) "Preferential replication of vaccinia virus in the ovaries is independent of immune regulation through IL-10 and TGF-." Author(s):Zhao Y,Adams YF,Croft M PubMed Article URL: was used in Flow cytometry/cell sorting to show that mice that overexpress IL-27 had decreased Treg frequencies and developed spontaneous inflammation. Journal of immunology (Baltimore, Md. : 1950) (Jul 2011; 187: 266) "A role for IL-27 in limiting T regulatory cell populations." Author(s):Wojno ED,Hosken N,Stumhofer JS,O'Hara AC,Mauldin E,Fang Q,Turka LA,Levin SD,Hunter CA PubMed Article URL: was used in Flow cytometry/cell sorting to study the immunological effects of IDO in -galactosylceramide (-GalCer)- induced hepatitis. Journal of immunology (Baltimore, Md. : 1950) (Oct 2010; 185: 4554) "Ability of IDO to attenuate liver injury in alpha-galactosylceramide-induced hepatitis model." Author(s):Ito H,Hoshi M,Ohtaki H,Taguchi A,Ando K,Ishikawa T,Osawa Y,Hara A,Moriwaki H,Saito K,Seishima M PubMed Article URL: was used in Flow cytometry/cell sorting to demonstrate dramatic differences in host T cell responses to noninflammatory Ags compared with responses to pathogens. 2 Immunocytochemistry References Journal of immunology (Baltimore, Md. : 1950) (Oct 2007; 179: 4988) "The surprising kinetics of the T cell response to live antigenic cells." Author(s):Tyznik AJ,Bevan MJ PubMed Article URL: was used in Immunocytochemistry to investigate whether the two vaccine components adenovirus and modified vaccinia virus Ankara can be co-injected to enhance protection against malaria. Molecular therapy : the journal of the American Society of Gene Therapy (Aug 2012; 20: 1633) "Mixed vector immunization with recombinant adenovirus and MVA can improve vaccine efficacy while decreasing antivector immunity." Author(s):Reyes-Sandoval A,Rollier CS,Milicic A,Bauza K,Cottingham MG,Tang CK,Dicks MD,Wang D,Longley RJ,Wyllie DH, Hill AV PubMed Article URL:

6 was used in Flow cytometry/cell sorting to study the immunological effects of IDO in -galactosylceramide (-GalCer)- induced hepatitis. 1 ELISA References Journal of immunology (Baltimore, Md. : 1950) (Oct 2010; 185: 4554) "Ability of IDO to attenuate liver injury in alpha-galactosylceramide-induced hepatitis model." Author(s):Ito H,Hoshi M,Ohtaki H,Taguchi A,Ando K,Ishikawa T,Osawa Y,Hara A,Moriwaki H,Saito K,Seishima M PubMed Article URL: was used in an ELISA assay to investigate TLR9 and MyD88 regulation of pro/anti-inflammatory cytokines in mice infected with Plasmodium yoelii. 1 Immunofluorescence References Journal of immunology (Baltimore, Md. : 1950) (May 2012; 188: 5073) "TLR9 and MyD88 are crucial for the development of protective immunity to malaria." Author(s):Gowda NM,Wu X,Gowda DC PubMed Article URL: 2 Immunohistochemistry (Frozen) References was used in Immunofluorescence to reveal that costimulatory pathways can be targeted to modulate the magnitude of an adoptive immunotherapeutic regimen. Journal of virology (Sep 2009; 83: 8905) "Therapeutic memory T cells require costimulation for effective clearance of a persistent viral infection." Author(s):Garidou L,Heydari S,Truong P,Brooks DG,McGavern DB PubMed Article URL: 1 Immunohistochemistry References Published figure using TNF alpha monoclonal antibody (Product # ) in Immunohistochemistry The Journal of clinical investigation (Aug 2001; 108: 415) "Conjugation of a self-antigen to papillomavirus-like particles allows for efficient induction of protective autoantibodies." Author(s):Chackerian B,Lowy DR,Schiller JT PubMed Article URL: Arthritis and rheumatism (Oct 1998; 41: 1806) "Therapeutic actions of cyclosporine and anti-tumor necrosis factor alpha in collagen-induced arthritis and the effect of combination therapy." Author(s):Williams RO,Mauri C,Mason LJ,Marinova-Mutafchieva L,Ross SE,Feldmann M,Maini RN PubMed Article URL: 1 Miscellaneous PubMed References 1 Functional Assay References Published figure using TNF alpha monoclonal antibody (Product # ) in Immunohistochemistry The Journal of clinical investigation (Aug 2001; 108: 415) "Conjugation of a self-antigen to papillomavirus-like particles allows for efficient induction of protective autoantibodies." Author(s):Chackerian B,Lowy DR,Schiller JT PubMed Article URL: Published figure using TNF alpha monoclonal antibody (Product # ) in Immunohistochemistry The Journal of clinical investigation (Aug 2001; 108: 415) "Conjugation of a self-antigen to papillomavirus-like particles allows for efficient induction of protective autoantibodies." Author(s):Chackerian B,Lowy DR,Schiller JT PubMed Article URL:

7 Arthritis and rheumatism (Oct 1998; 41: 1806) "Therapeutic actions of cyclosporine and anti-tumor necrosis factor alpha in collagen-induced arthritis and the effect of combination therapy." Author(s):Williams RO,Mauri C,Mason LJ,Marinova-Mutafchieva L,Ross SE,Feldmann M,Maini RN PubMed Article URL: