Antibody data
- Antibody Data
- Antigen structure
- References [20]
- Comments [0]
- Validations
- Other assay [1]
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- Product number
- M303 - Provider product page
- Provider
- Invitrogen Antibodies
- Product name
- TNF alpha Monoclonal Antibody (2TNF-H34A)
- Antibody type
- Monoclonal
- Antigen
- Recombinant full-length protein
- Description
- M303 targets TNF Alpha in ELISA, and WB applications and shows reactivity with Human samples. The M303 antibody (clone 2TNF-H34A) has successfully been paired as the coating antibody in a sandwich ELISA with detection antibody M302B (biotinylated conjugate of clone 2TNG-H33). The M303 immunogen is recombinant human TNF alpha. M303 detects TNF Alpha which has a predicted molecular weight of approximately 26 kDa. The M303 TNFa antibody (clone 2TNF-H34A) has successfully been paired as the coating antibody in a sandwich ELISA with detection antibody M302B (biotinylated conjugate of clone 2TNF-H33) Typical dilutions for sandwich ELISA include 1 µg/mL for coating and 0.125 - 0.25 µg/mL for detection. Antibody M303 (clone 2TNF-H34A) and biotinylated antibody M302B (clone 2TNF-H33) have successfully been used in combination with recombinant TNFa protein STNFA in ELISA applications. This product has been tested for endotoxins by limulus amoebocyte lysate (LAL) assay and contains an endotoxin concentration of less than or equal to 10 endotoxin units per milligram (EU/mg).
- Reactivity
- Human
- Host
- Mouse
- Isotype
- IgG
- Antibody clone number
- 2TNF-H34A
- Vial size
- 500 µg
- Concentration
- 1.0 mg/mL
- Storage
- -20°C
Submitted references Protein kinase cα regulates the expression of complement receptor Ig in human monocyte-derived macrophages.
Reduced frequency of a CD14+ CD16+ monocyte subset with high Toll-like receptor 4 expression in cord blood compared to adult blood contributes to lipopolysaccharide hyporesponsiveness in newborns.
Anandamide suppresses proliferation and cytokine release from primary human T-lymphocytes mainly via CB2 receptors.
Cytokines across the night in chronic fatigue syndrome with and without fibromyalgia.
Efficient immobilization of a ligand antibody with high antigen-binding activity by use of a polystyrene-binding peptide and an intelligent microtiter plate.
Sesquiterpene lactones induce distinct forms of cell death that modulate human monocyte-derived macrophage responses.
Protein microarray platform for the multiplex analysis of biomarkers in human sera.
The immunostimulant RU41740 from Klebsiella pneumoniae activates human cells in whole blood to potentially stimulate innate and adaptive immune responses.
Thalidomide-based TNF-alpha inhibitors for neurodegenerative diseases.
Generation of three-dimensional pannus-like tissues in vitro from single cell suspensions of synovial fluid cells from arthritis patients.
Host factors impacting the innate response in humans to the candidate adjuvants RC529 and monophosphoryl lipid A.
Long-term treatment with etanercept significantly reduces the number of proinflammatory cytokine-secreting peripheral blood mononuclear cells in patients with rheumatoid arthritis.
Activation of influenza virus-specific CD4+ and CD8+ T cells: a new role for plasmacytoid dendritic cells in adaptive immunity.
Enhanced monocyte Th1 cytokine production in HCV-infected cryoglobulinemic patients.
Enhanced monocyte Th1 cytokine production in HCV-infected cryoglobulinemic patients.
Interferon--gamma control of EBV-transformed B cells: a role for CD8+ T cells that poorly kill EBV-infected cells.
Enhanced interleukin-12 and CD40 ligand activities but reduced Staphylococcus aureus Cowan 1-induced responses suggest a generalized and progressively impaired type 1 cytokine pattern for human schistosomiasis.
Enhanced interleukin-12 and CD40 ligand activities but reduced Staphylococcus aureus Cowan 1-induced responses suggest a generalized and progressively impaired type 1 cytokine pattern for human schistosomiasis.
Regulation of interleukin-12 by complement receptor 3 signaling.
Regulation of interleukin-12 by complement receptor 3 signaling.
Ma Y, Usuwanthim K, Munawara U, Quach A, Gorgani NN, Abbott CA, Hii CS, Ferrante A
Journal of immunology (Baltimore, Md. : 1950) 2015 Mar 15;194(6):2855-61
Journal of immunology (Baltimore, Md. : 1950) 2015 Mar 15;194(6):2855-61
Reduced frequency of a CD14+ CD16+ monocyte subset with high Toll-like receptor 4 expression in cord blood compared to adult blood contributes to lipopolysaccharide hyporesponsiveness in newborns.
Pedraza-Sánchez S, Hise AG, Ramachandra L, Arechavaleta-Velasco F, King CL
Clinical and vaccine immunology : CVI 2013 Jul;20(7):962-71
Clinical and vaccine immunology : CVI 2013 Jul;20(7):962-71
Anandamide suppresses proliferation and cytokine release from primary human T-lymphocytes mainly via CB2 receptors.
Cencioni MT, Chiurchiù V, Catanzaro G, Borsellino G, Bernardi G, Battistini L, Maccarrone M
PloS one 2010 Jan 14;5(1):e8688
PloS one 2010 Jan 14;5(1):e8688
Cytokines across the night in chronic fatigue syndrome with and without fibromyalgia.
Nakamura T, Schwander SK, Donnelly R, Ortega F, Togo F, Broderick G, Yamamoto Y, Cherniack NS, Rapoport D, Natelson BH
Clinical and vaccine immunology : CVI 2010 Apr;17(4):582-7
Clinical and vaccine immunology : CVI 2010 Apr;17(4):582-7
Efficient immobilization of a ligand antibody with high antigen-binding activity by use of a polystyrene-binding peptide and an intelligent microtiter plate.
Kumada Y, Hamasaki K, Shiritani Y, Ohse T, Kishimoto M
Journal of biotechnology 2009 Jun 15;142(2):135-41
Journal of biotechnology 2009 Jun 15;142(2):135-41
Sesquiterpene lactones induce distinct forms of cell death that modulate human monocyte-derived macrophage responses.
López-Antón N, Hermann C, Murillo R, Merfort I, Wanner G, Vollmar AM, Dirsch VM
Apoptosis : an international journal on programmed cell death 2007 Jan;12(1):141-53
Apoptosis : an international journal on programmed cell death 2007 Jan;12(1):141-53
Protein microarray platform for the multiplex analysis of biomarkers in human sera.
Urbanowska T, Mangialaio S, Zickler C, Cheevapruk S, Hasler P, Regenass S, Legay F
Journal of immunological methods 2006 Oct 20;316(1-2):1-7
Journal of immunological methods 2006 Oct 20;316(1-2):1-7
The immunostimulant RU41740 from Klebsiella pneumoniae activates human cells in whole blood to potentially stimulate innate and adaptive immune responses.
Pedraza-Sánchez S, González-Hernández Y, Escobar-Gutiérrez A, Ramachandra L
International immunopharmacology 2006 Apr;6(4):635-46
International immunopharmacology 2006 Apr;6(4):635-46
Thalidomide-based TNF-alpha inhibitors for neurodegenerative diseases.
Greig NH, Giordano T, Zhu X, Yu QS, Perry TA, Holloway HW, Brossi A, Rogers JT, Sambamurti K, Lahiri DK
Acta neurobiologiae experimentalis 2004;64(1):1-9
Acta neurobiologiae experimentalis 2004;64(1):1-9
Generation of three-dimensional pannus-like tissues in vitro from single cell suspensions of synovial fluid cells from arthritis patients.
Solomon S, Masilamani M, Mohanty S, Schwab JE, Boneberg EM, Illges H
Rheumatology international 2004 Mar;24(2):71-6
Rheumatology international 2004 Mar;24(2):71-6
Host factors impacting the innate response in humans to the candidate adjuvants RC529 and monophosphoryl lipid A.
Tiberio L, Fletcher L, Eldridge JH, Duncan DD
Vaccine 2004 Mar 29;22(11-12):1515-23
Vaccine 2004 Mar 29;22(11-12):1515-23
Long-term treatment with etanercept significantly reduces the number of proinflammatory cytokine-secreting peripheral blood mononuclear cells in patients with rheumatoid arthritis.
Schotte H, Schlüter B, Willeke P, Mickholz E, Schorat MA, Domschke W, Gaubitz M
Rheumatology (Oxford, England) 2004 Aug;43(8):960-4
Rheumatology (Oxford, England) 2004 Aug;43(8):960-4
Activation of influenza virus-specific CD4+ and CD8+ T cells: a new role for plasmacytoid dendritic cells in adaptive immunity.
Fonteneau JF, Gilliet M, Larsson M, Dasilva I, Münz C, Liu YJ, Bhardwaj N
Blood 2003 May 1;101(9):3520-6
Blood 2003 May 1;101(9):3520-6
Enhanced monocyte Th1 cytokine production in HCV-infected cryoglobulinemic patients.
Loffreda S, Muratori P, Muratori L, Mele L, Bianchi FB, Lenzi M
Journal of hepatology 2003 Feb;38(2):230-6
Journal of hepatology 2003 Feb;38(2):230-6
Enhanced monocyte Th1 cytokine production in HCV-infected cryoglobulinemic patients.
Loffreda S, Muratori P, Muratori L, Mele L, Bianchi FB, Lenzi M
Journal of hepatology 2003 Feb;38(2):230-6
Journal of hepatology 2003 Feb;38(2):230-6
Interferon--gamma control of EBV-transformed B cells: a role for CD8+ T cells that poorly kill EBV-infected cells.
Shi Y, Lutz CT
Viral immunology 2002;15(1):213-25
Viral immunology 2002;15(1):213-25
Enhanced interleukin-12 and CD40 ligand activities but reduced Staphylococcus aureus Cowan 1-induced responses suggest a generalized and progressively impaired type 1 cytokine pattern for human schistosomiasis.
Montenegro SM, Abath FG, Domingues AL, Melo WG, Morais CN, Coutinho EM, Mahanty S, Wynn TA
Infection and immunity 2002 Nov;70(11):5903-12
Infection and immunity 2002 Nov;70(11):5903-12
Enhanced interleukin-12 and CD40 ligand activities but reduced Staphylococcus aureus Cowan 1-induced responses suggest a generalized and progressively impaired type 1 cytokine pattern for human schistosomiasis.
Montenegro SM, Abath FG, Domingues AL, Melo WG, Morais CN, Coutinho EM, Mahanty S, Wynn TA
Infection and immunity 2002 Nov;70(11):5903-12
Infection and immunity 2002 Nov;70(11):5903-12
Regulation of interleukin-12 by complement receptor 3 signaling.
Marth T, Kelsall BL
The Journal of experimental medicine 1997 Jun 2;185(11):1987-95
The Journal of experimental medicine 1997 Jun 2;185(11):1987-95
Regulation of interleukin-12 by complement receptor 3 signaling.
Marth T, Kelsall BL
The Journal of experimental medicine 1997 Jun 2;185(11):1987-95
The Journal of experimental medicine 1997 Jun 2;185(11):1987-95
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Supportive validation
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- Invitrogen Antibodies (provider)
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- Experimental details
- Direct ELISA analysis of TNF alpha was performed by coating wells of a 96-well plate with 100 µL per well of human recombinant protein TNF alpha (Product # RTNFAI) diluted in carbonate/bicarbonate buffer (Product # 28382) at a concentration of 1 µg/mL overnight at 4C. Wells of the plate were washed, blocked with starting blocking buffer (Product # 37538), and incubated with 100 µL per well of a mouse anti-TNF alpha monoclonal antibody (Product # M303) at a dilution of 1:50, 1:100, 1:200, 1:400, 1:800, 1:1600 and 1:3200 for 90 minutes at 37C. The plate was washed, then incubated with 100 µL per well of a HRP-conjugated goat anti-mouse IgG secondary antibody (Product # 62-6520) at a dilution of 1:2000 for 90 minutes at 37C. Detection was performed using 1-Step Ultra TMB substrate (Product # 34028) for 5-10 minutes at room temperature in the dark. The reaction was stopped with Stop solution (Product # N600), and absorbances were read on a spectrophotometer at 450-550 nm.