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Direct ELISA analysis of TNF alpha was performed by coating wells of a 96-well plate with 100ul per well of human recombinant protein TNF alpha (Product #RTNFAI) diluted in carbonate/bicarbonate buffer (Product # 28382) at a concentration of 1ug/ml overnight at 4C. Wells of the plate were washed, blocked with starting blocking buffer (Product # 37538), and incubated with 100ul per well of a rabbit anti-TNF alpha polyclonal antibody (Product # M302) at a dilution of 1:25, 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 100ul per well of a HRP-conjugated goat anti-rabbit IgG secondary antibody (Product # 65-6120) 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 450-550nm.
|Tested species reactivity||Human|
|Published species reactivity||Rabbit, Rat, Human, Mouse|
|Host / Isotype||Rabbit / IgG|
|Immunogen||Recombinant human TNF alpha|
|Tested Applications||Dilution *|
|Immunohistochemistry (Paraffin) (IHC (P))||Assay Dependent|
|Neutralization (Neu)||Assay Dependent|
|Western Blot (WB)||Assay Dependent|
* 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.
P300A targets TNF Alpha in ELISA, IHC(P), Neu, and WB applications and shows reactivity with Human samples.
The P300A immunogen is recombinant human TNF alpha.
P300A detects TNF Alpha which has a predicted molecular weight of approximately 26 kDa.
This gene encodes a multifunctional proinflammatory cytokine that belongs to the tumor necrosis factor (TNF) superfamily. This cytokine is mainly secreted by macrophages. It can bind to, and thus functions through its receptors TNFRSF1A/TNFR1 and TNFRSF1B/TNFBR. This cytokine is involved in the regulation of a wide spectrum of biological processes including cell proliferation, differentiation, apoptosis, lipid metabolism, and coagulation. This cytokine has been implicated in a variety of diseases, including autoimmune diseases, insulin resistance, and cancer. Knockout studies in mice also suggested the neuroprotective function of this cytokine.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.
HIV-1 inhibits autophagy in bystander macrophage/monocytic cells through Src-Akt and STAT3.
P300A was used in blocking or activating experiment to investigate the mechanism of the inhibitory effect of HIV-1 on autophagy
|Van Grol J,Subauste C,Andrade RM,Fujinaga K,Nelson J,Subauste CS||PloS one (5:null)||2010|
Cyclic AMP increases endogenous granulocyte colony-stimulating factor formation in monocytes and THP-1 macrophages despite attenuated TNF-alpha formation.
P300A was used in blocking or activating experiment to study whether drugs that suppress TNF-alpha also impair G-CSF production
|Hareng L,Meergans T,von Aulock S,Volk HD,Hartung T||European journal of immunology (33:2287)||2003|
A model of human whole blood lymphokine release for in vitro and ex vivo use.
P300A was used in blocking or activating experiment to develop a model of human whole blood lymphokine release for in vitro and ex vivo use
|Hermann C,von Aulock S,Graf K,Hartung T||Journal of immunological methods (275:69)||2003|
Monocyte:astrocyte interactions regulate MCP-1 expression in both cell types.
P300A was used in blocking or activating experiment to study the role of monocyte-astrocyte interactions in regulating MCP-1 expression in both cell types
|Andjelkovic AV,Kerkovich D,Pachter JS||Journal of leukocyte biology (68:545)||2000|
Activation of nuclear factor-kappa B and gene expression in human endothelial cells by the common haptens nickel and cobalt.
P300A was used in blocking/activating experiment to investigate the effect of nickel chloride and cobalt chloride on the expression of NF-kappa B in human endothelial cells
|Goebeler M,Roth J,Bröcker EB,Sorg C,Schulze-Osthoff K||Journal of immunology (Baltimore, Md. : 1950) (155:2459)||1995|
Soluble IgA and IgG aggregates are catabolized by cultured rat mesangial cells and induce production of TNF-alpha and IL-6, and proliferation.
P300A was used in blocking/activating experiment to study the mechanism for the mesangial deposits of IgA immune complexes in IgA nephropathy
|Gómez-Guerrero C,López-Armada MJ,González E,Egido J||Journal of immunology (Baltimore, Md. : 1950) (153:5247)||1994|
Tumour necrosis factor production by IL-2-activated macrophages in vitro and in vivo.
P300A was used in blocking/activating experiment to investigate the expression of tumor necrosis factor by macrophages through interleukin 2 activation
|Economou JS,McBride WH,Essner R,Rhoades K,Golub S,Holmes EC,Morton DL||Immunology (67:514)||1989|
Tumor necrosis factor-related apoptosis-inducing ligand induces neuronal death in a murine model of HIV central nervous system infection.
P300A was used in immunohistochemistry to investigate the role of TRAIL expressed in HIV-1-infected macrophages on neuronal apoptosis
|Miura Y,Misawa N,Kawano Y,Okada H,Inagaki Y,Yamamoto N,Ito M,Yagita H,Okumura K,Mizusawa H,Koyanagi Y||Proceedings of the National Academy of Sciences of the United States of America (100:2777)||2003|
Localization of cytokines in cholesteatoma tissue.
P300A was used in immunohistochemistry to investigate the distribution of various cytokines in cholesteatoma tissue
|Marenda SA,Aufdemorte TB||Otolaryngology--head and neck surgery : official journal of American Academy of Otolaryngology-Head and Neck Surgery (112:359)||1995|
Inhibition of TNF-alpha processing and TACE-mediated ectodomain shedding by ethanol.
P300A was used in immunocytochemistry to study the inhibitory effect of ethanol on TNF-alpha processing and TACE-mediated ectodomain shedding in monocytic cell lines
|Zhang Z,Cork J,Ye P,Lei D,Schwarzenberger PO,Summer WR,Shellito JE,Nelson S,Kolls JK||Journal of leukocyte biology (67:856)||2000|
Modified natural porcine surfactant inhibits superoxide anions and proinflammatory mediators released by resting and stimulated human monocytes.
P300A was used in ELISA to investigate the effect of modified natural porcine surfactant on the stimulation of monocytes
|Walti H,Polla BS,Bachelet M||Pediatric research (41:114)||1997|
Enhancement of fibronectin-induced migration of corneal epithelial cells by cytokines.
P300A was used in ELISA to investigate the effect of cytokines on corneal epithelial cell migration
|Wang X,Kamiyama K,Iguchi I,Kita M,Imanishi J||Investigative ophthalmology and visual science (35:4001)||1994|
Role of IL-1 and tumor necrosis factor in coxsackie virus-induced autoimmune myocarditis.
P300A was used in ELISA to investigate the importance of interleukin 1 and tumor necrosis factor in the development of autoimmune myocarditis
|Lane JR,Neumann DA,Lafond-Walker A,Herskowitz A,Rose NR||Journal of immunology (Baltimore, Md. : 1950) (151:1682)||1993|
APC1 protein; Cachectin; TNF superfamily, member 2; TNF, macrophage-derived; TNF, monocyte-derived; TNF-a; TNF-alpha; Tumor necrosis factor; tumor necrosis factor alpha; tumor necrosis factor ligand 1F; Tumor necrosis factor ligand superfamily member 2; tumor necrosis factor-alpha
DIF; TNF; TNF-alpha; TNFA; TNFSF2; TNLG1F