Sandwich ELISA analysis of mouse TNF-alpha was performed using a Mouse TNF alpha Colorimetric ELISA kit (Product # EMTNFA) by loading 50 ul per well of Mouse TNF-alpha Recombinant Protein (Product # SMTNFA) in dodecuplicate at 2450, 350, 50, and 0 pg/ml across a 3 ug/ml rat anti-mouse TNF-alpha (Product # MM350C ) pre-coated plate and incubating for 2 hours at room temperature along with 50 ul per well of rat anti-mouse TNF-alpha biotinylated antibody (Product # MM350DB). The plate was washed and incubated with 100 ul per well of Streptavidin-HRP (Product # N504) in all test wells at 1:30,000 for 30 minutes at room temperature. Detection was performed using 1-Step Ultra TMB Substrate (Product # 34028) for 30 minutes at room temperature in the dark. The plate was then stopped with 0.16M sulfuric acid. Absorbances were read on a spectrophotometer at 450-550 nm.
|Tested species reactivity||Mouse|
|Published species reactivity||Mouse, Not Applicable|
|Host / Isotype||Rat / IgG1|
|Immunogen||Recombinant mouse TNF alpha|
|Purification||Ammonium sulfate precipitation|
|Tested Applications||Dilution *|
|ELISA (ELISA)||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.
MM350C targets TNF Alpha in ELISA, and WB applications and shows reactivity with mouse samples.
The MM350C immunogen is recombinant mouse TNF alpha. This antibody is produced by injecting Rat IgG secreting hybridoma cells into the peritoneum of mice. The resulting ascites is collected from the mice and the antibody is purified.
MM350C detects TNF Alpha which has a predicted molecular weight of approximately 26 kDa.
The MM350C mouse TNFa antibody (clone XT3) has successfully been paired as the coating antibody in a sandwich ELISA with detection antibody MM350DB (biotinylated conjugate of clone XT22). Typical dilutions for sandwich ELISA include 1 µg/ml for coating and 0.125 - 0.25 µg/ml for detection.
Antibody MM350C (clone XT3) and biotinylated antibody MM350DB (clone XT22) have successfully been used in combination with recombinant mouse TNFa protein SMTNFA in ELISA applications.
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.
Inhibition of endoplasmic reticulum stress alleviates lipopolysaccharide-induced lung inflammation through modulation of NF-¿B/HIF-1¿ signaling pathway.
MM350C was used in western blot to study the mechanism by which inhibition of endoplasmic reticulum stress reduces lung inflammation induced by LPS
|Kim HJ,Jeong JS,Kim SR,Park SY,Chae HJ,Lee YC||Scientific reports (3:null)||2013|
Interleukin-1beta and tumor necrosis factor-alpha are expressed by different subsets of microglia and macrophages after ischemic stroke in mice.
MM350C was used in immunohistochemistry to test if IL-1beta and TNF-alpha are synthesized by overlapping or segregated populations of cells after ischemic stroke in mice
|Clausen BH,Lambertsen KL,Babcock AA,Holm TH,Dagnaes-Hansen F,Finsen B||Journal of neuroinflammation (5:null)||2008|
Cyclooxygenase-2 inhibition improves amyloid-beta-mediated suppression of memory and synaptic plasticity.
MM350C was used in ELISA to study the role of COX-2 inhibition in amyloid-beta-mediated suppression of LTP and memory function
|Kotilinek LA,Westerman MA,Wang Q,Panizzon K,Lim GP,Simonyi A,Lesne S,Falinska A,Younkin LH,Younkin SG,Rowan M,Cleary J,Wallis RA,Sun GY,Cole G,Frautschy S,Anwyl R,Ashe KH||Brain : a journal of neurology (131:651)||2008|
Inhibition of vagally mediated immune-to-brain signaling by vanadyl sulfate speeds recovery from sickness.
MM350C was used in ELISA to investigate the role of TLR2 in the interferon-gamma responses
|Johnson DR,O'Connor JC,Dantzer R,Freund GG||Proceedings of the National Academy of Sciences of the United States of America (102:15184)||2005|
|Not Applicable||Not Cited||
Redirecting in vivo elicited tumor infiltrating macrophages and dendritic cells towards tumor rejection.
MM350C was used in ELISA to describe the mechanisms by which tumor-infiltrating macrophages and dendritic cells can be redirected to become potent effectors and activators of the innate and adaptive immunity
|Guiducci C,Vicari AP,Sangaletti S,Trinchieri G,Colombo MP||Cancer research (65:3437)||2005|
Temporal correlation of tumor necrosis factor-alpha release, upregulation of pulmonary ICAM-1 and VCAM-1, neutrophil sequestration, and lung injury in diet-induced pancreatitis.
MM350C was used in ELISA to investigate the interplay among tumor necrosis factor alpha, ICAM-1, and VCAM-1 in diet-induced pancreatitis
|Lundberg AH,Granger N,Russell J,Callicutt S,Gaber LW,Kotb M,Sabek O,Gaber AO||Journal of gastrointestinal surgery : official journal of the Society for Surgery of the Alimentary Tract (4:248)||2000|
Immunization of mice with phosphatidylcholine drastically reduces the parasitaemia of subsequent Plasmodium chabaudi chabaudi blood-stage infections.
MM350C was used in ELISA to investigate the effect of phosphatidylcholine immunization of mice for the parasitaemia of subsequent Plasmodium chabaudi chabaudi blood-stage infections
|Bordmann G,Rudin W,Favre N||Immunology (94:35)||1998|
Lipopolysaccharide-induced osteoclastogenesis in Src homology 2-domain phosphatase-1-deficient viable motheaten mice.
MM350C was used in blocking/activating experiment to study the mechanism for lipopolysaccharide-induced osteoclastogenesis
|Hayashi S,Tsuneto M,Yamada T,Nose M,Yoshino M,Shultz LD,Yamazaki H||Endocrinology (145:2721)||2004|
Distinct osteoclast precursors in the bone marrow and extramedullary organs characterized by responsiveness to Toll-like receptor ligands and TNF-alpha.
MM350C was used in blocking/activating experiment to study the functions of TLR ligands and tumor necross factor alpha in osteoclast precursors in bone marrow and extramedullary organs
|Hayashi S,Yamada T,Tsuneto M,Yamane T,Takahashi M,Shultz LD,Yamazaki H||Journal of immunology (Baltimore, Md. : 1950) (171:5130)||2003|
TNF regulates thymocyte production by apoptosis and proliferation of the triple negative (CD3-CD4-CD8-) subset.
MM350C was used in blocking/activating experiment to study the tumor necrosis factor regulated homeostasis of thymocyte production by TN (CD3-CD4-CD8-) subset apoptosis and proliferation
|Baseta JG,Stutman O||Journal of immunology (Baltimore, Md. : 1950) (165:5621)||2000|
Nitric oxide-producing CD11b(+)Ly-6G(Gr-1)(+)CD31(ER-MP12)(+) cells in the spleen of cyclophosphamide-treated mice: implications for T-cell responses in immunosuppressed mice.
MM350C was used in blocking/activating experiment to investigate the mechanism for cyclophosphamide-induced immunosuppression
|Angulo I,de las Heras FG,García-Bustos JF,Gargallo D,Muñoz-Fernández MA,Fresno M||Blood (95:212)||2000|