Western Blot (WB)
(Figure 1. Western blot analysis of TNF Alpha/Tnf using anti-TNF Alpha/Tnf antibody (MBS1753257).Electrophoresis was performed on a 5-20% SDS-PAGE gel at 70V (Stacking gel) / 90V (Resolving gel) for 2-3 hours. The sample well of each lane was loaded with 50ug of sample under reducing conditions.Lane 1: rat thymus tissue lysatesLane 2: mouse thymus tissue lysates.After Electrophoresis, proteins were transferred to a Nitrocellulose membrane at 150mA for 50-90 minutes. Blocked the membrane with 5% Non-fat Milk/ TBS for 1. 5 hour at RT. The membrane was incubated with rabbit anti-TNF Alpha/Tnf antigen affinity purified polyclonal antibody (Catalog # MBS1753257) at 0. 5 μg/mL overnight at 4 degree C, then washed with TBS-0. 1%Tween 3 times with 5 minutes each and probed with a goat anti-rabbit IgG-HRP secondary antibody at a dilution of 1:5000 for 1. 5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit (Catalog # MBS176460) with Tanon 5200 system. A specific band was detected for TNF Alpha/Tnf at approximately 25KD. The expected band size for TNF Alpha/Tnf is at 25KD. )
Rabbit anti-Mouse, Rat TNF alpha/Tnf Polyclonal Antibody | anti-Tnf antibody
Anti-TNF alpha/Tnf Antibody
ICC/IF: 5ug/ml|Mouse|
FC/FACS/FCM: 1-3ug/1x106 cells|Mouse|
Direct ELISA: 0.1-0.5ug/ml|Mouse|
Western Blot (WB)
(Figure 1. Western blot analysis of TNF Alpha/Tnf using anti-TNF Alpha/Tnf antibody (MBS1753257).Electrophoresis was performed on a 5-20% SDS-PAGE gel at 70V (Stacking gel) / 90V (Resolving gel) for 2-3 hours. The sample well of each lane was loaded with 50ug of sample under reducing conditions.Lane 1: rat thymus tissue lysatesLane 2: mouse thymus tissue lysates.After Electrophoresis, proteins were transferred to a Nitrocellulose membrane at 150mA for 50-90 minutes. Blocked the membrane with 5% Non-fat Milk/ TBS for 1. 5 hour at RT. The membrane was incubated with rabbit anti-TNF Alpha/Tnf antigen affinity purified polyclonal antibody (Catalog # MBS1753257) at 0. 5 μg/mL overnight at 4 degree C, then washed with TBS-0. 1%Tween 3 times with 5 minutes each and probed with a goat anti-rabbit IgG-HRP secondary antibody at a dilution of 1:5000 for 1. 5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit (Catalog # MBS176460) with Tanon 5200 system. A specific band was detected for TNF Alpha/Tnf at approximately 25KD. The expected band size for TNF Alpha/Tnf is at 25KD. )
Western Blot (WB)
(Figure 1. Western blot analysis of TNF Alpha/Tnf using anti-TNF Alpha/Tnf antibody (MBS1753257).Electrophoresis was performed on a 5-20% SDS-PAGE gel at 70V (Stacking gel) / 90V (Resolving gel) for 2-3 hours. The sample well of each lane was loaded with 50ug of sample under reducing conditions.Lane 1: rat thymus tissue lysatesLane 2: mouse thymus tissue lysates.After Electrophoresis, proteins were transferred to a Nitrocellulose membrane at 150mA for 50-90 minutes. Blocked the membrane with 5% Non-fat Milk/ TBS for 1. 5 hour at RT. The membrane was incubated with rabbit anti-TNF Alpha/Tnf antigen affinity purified polyclonal antibody (Catalog # MBS1753257) at 0. 5 μg/mL overnight at 4 degree C, then washed with TBS-0. 1%Tween 3 times with 5 minutes each and probed with a goat anti-rabbit IgG-HRP secondary antibody at a dilution of 1:5000 for 1. 5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit (Catalog # MBS176460) with Tanon 5200 system. A specific band was detected for TNF Alpha/Tnf at approximately 25KD. The expected band size for TNF Alpha/Tnf is at 25KD. )
Immunofluorescence (IF)
(Figure 2. IF analysis of TNF Alpha/Tnf using anti- TNF Alpha/Tnf antibody (MBS1753257).TNF Alpha/Tnf was detected in immunocytochemical section of HEPA1-6 cells. Enzyme antigen retrieval was performed using IHC enzyme antigen retrieval reagent (MBS176582) for 15 mins. The cells were blocked with 10% goat serum. And then incubated with 5μg/mL rabbit anti- TNF Alpha/Tnf Antibody (MBS1753257) overnight at 4 degree C. DyLight®594 Conjugated Goat Anti-Rabbit IgG (BA1142) was used as secondary antibody at 1:100 dilution and incubated for 30 minutes at 37 degree C. The section was counterstained with DAPI. Visualize using a fluorescence microscope and filter sets appropriate for the label used. )
Immunofluorescence (IF)
(Figure 2. IF analysis of TNF Alpha/Tnf using anti- TNF Alpha/Tnf antibody (MBS1753257).TNF Alpha/Tnf was detected in immunocytochemical section of HEPA1-6 cells. Enzyme antigen retrieval was performed using IHC enzyme antigen retrieval reagent (MBS176582) for 15 mins. The cells were blocked with 10% goat serum. And then incubated with 5μg/mL rabbit anti- TNF Alpha/Tnf Antibody (MBS1753257) overnight at 4 degree C. DyLight®594 Conjugated Goat Anti-Rabbit IgG (BA1142) was used as secondary antibody at 1:100 dilution and incubated for 30 minutes at 37 degree C. The section was counterstained with DAPI. Visualize using a fluorescence microscope and filter sets appropriate for the label used. )
Flow Cytometry (FC/FACS)
(Figure 3. Flow Cytometry analysis of Raw264. 7 cells using anti-TNF Alpha/Tnf antibody (MBS1753257).Overlay histogram showing Raw264. 7 cells stained with MBS1753257 (Blue line). The cells were blocked with 10% normal goat serum. And then incubated with rabbit anti-TNF Alpha/Tnf Antibody (MBS1753257, 1μg/1x106 cells) for 30 min at 20 degree C. DyLight®488 conjugated goat anti-rabbit IgG (5-10μg/1x106 cells) was used as secondary antibody for 30 minutes at 20 degree C. Isotype control antibody (Green line) was rabbit IgG (1μg/1x106) used under the same conditions. Unlabelled sample (Red line) was also used as a control. )
Flow Cytometry (FC/FACS)
(Figure 3. Flow Cytometry analysis of Raw264. 7 cells using anti-TNF Alpha/Tnf antibody (MBS1753257).Overlay histogram showing Raw264. 7 cells stained with MBS1753257 (Blue line). The cells were blocked with 10% normal goat serum. And then incubated with rabbit anti-TNF Alpha/Tnf Antibody (MBS1753257, 1μg/1x106 cells) for 30 min at 20 degree C. DyLight®488 conjugated goat anti-rabbit IgG (5-10μg/1x106 cells) was used as secondary antibody for 30 minutes at 20 degree C. Isotype control antibody (Green line) was rabbit IgG (1μg/1x106) used under the same conditions. Unlabelled sample (Red line) was also used as a control. )
2. Shirai, T., Yamaguchi, H., Ito, H., Todd, C. W., Wallace, R. B. Cloning and expression in Escherichia coli of the gene for human tumour necrosis factor. Nature 313: 803-806, 1985.
NCBI and Uniprot Product Information
Uniprot Description
TNF-a: Cytokine that binds to TNFRSF1A/TNFR1 and TNFRSF1B/TNFBR. It is mainly secreted by macrophages and can induce cell death of certain tumor cell lines. It is potent pyrogen causing fever by direct action or by stimulation of interleukin-1 secretion and is implicated in the induction of cachexia, Under certain conditions it can stimulate cell proliferation and induce cell differentiation. Homotrimer. Interacts with SPPL2B. Belongs to the tumor necrosis factor family.
Protein type: Apoptosis; Cytokine; Membrane protein, integral; Motility/polarity/chemotaxis
Cellular Component: cell surface; external side of plasma membrane; extracellular space; integral to membrane; integral to plasma membrane; intracellular; lipid raft; phagocytic cup; plasma membrane; recycling endosome; secretory granule
Molecular Function: cytokine activity; identical protein binding; protease binding; protein binding; tumor necrosis factor receptor binding
Biological Process: activation of MAPK activity; activation of MAPKKK activity; activation of NF-kappaB transcription factor; apoptosis; calcium-mediated signaling; caspase activation; cell activation; cell proliferation; cellular extravasation; chronic inflammatory response to antigenic stimulus; cortical actin cytoskeleton organization and biogenesis; defense response; defense response to bacterium; defense response to Gram-positive bacterium; detection of mechanical stimulus involved in sensory perception of pain; DNA damage response, signal transduction resulting in induction of apoptosis; extracellular matrix organization and biogenesis; glucose metabolic process; humoral immune response; induction of apoptosis via death domain receptors; inflammatory response; JNK cascade; leukocyte migration; leukocyte tethering or rolling; lipopolysaccharide-mediated signaling pathway; MAPKKK cascade; multicellular organismal development; negative regulation of cell proliferation; negative regulation of cytokine secretion during immune response; negative regulation of endothelial cell proliferation; negative regulation of glucose import; negative regulation of interleukin-6 production; negative regulation of L-glutamate transport; negative regulation of lipid catabolic process; negative regulation of mitotic cell cycle; negative regulation of myelination; negative regulation of myoblast differentiation; negative regulation of osteoblast differentiation; negative regulation of protein complex disassembly; negative regulation of transcription from RNA polymerase II promoter; negative regulation of transcription, DNA-dependent; negative regulation of viral genome replication; organ morphogenesis; osteoclast differentiation; positive regulation of action potential; positive regulation of apoptosis; positive regulation of caspase activity; positive regulation of cell adhesion; positive regulation of cell proliferation; positive regulation of chemokine biosynthetic process; positive regulation of chemokine production; positive regulation of chronic inflammatory response to antigenic stimulus; positive regulation of cytokine production; positive regulation of cytokine secretion; positive regulation of fever; positive regulation of hair follicle development; positive regulation of heterotypic cell-cell adhesion; positive regulation of humoral immune response mediated by circulating immunoglobulin; positive regulation of I-kappaB kinase/NF-kappaB cascade; positive regulation of inflammatory response; positive regulation of interferon-gamma production; positive regulation of interleukin-18 production; positive regulation of interleukin-6 production; positive regulation of interleukin-8 biosynthetic process; positive regulation of interleukin-8 production; positive regulation of JNK activity; positive regulation of JNK cascade; positive regulation of MAP kinase activity; positive regulation of membrane protein ectodomain proteolysis; positive regulation of mitosis; positive regulation of neuron apoptosis; positive regulation of NF-kappaB import into nucleus; positive regulation of NFAT protein import into nucleus; positive regulation of nitric oxide biosynthetic process; positive regulation of osteoclast differentiation; positive regulation of peptidyl-serine phosphorylation; positive regulation of phagocytosis; positive regulation of programmed cell death; positive regulation of protein amino acid phosphorylation; positive regulation of protein complex assembly; positive regulation of protein complex disassembly; positive regulation of protein kinase activity; positive regulation of protein kinase B signaling cascade; positive regulation of protein transport; positive regulation of smooth muscle cell proliferation; positive regulation of synaptic transmission; positive regulation of transcription factor activity; positive regulation of transcription from RNA polymerase II promoter; positive regulation of transcription, DNA-dependent; positive regulation of translational initiation by iron; protein import into nucleus, translocation; protein kinase B signaling cascade; receptor biosynthetic process; regulation of cell proliferation; regulation of I-kappaB kinase/NF-kappaB cascade; regulation of immunoglobulin secretion; regulation of insulin secretion; regulation of osteoclast differentiation; regulation of protein amino acid phosphorylation; regulation of protein secretion; response to glucocorticoid stimulus; response to lipopolysaccharide; response to organic substance; response to virus; sequestering of triacylglycerol; signal transduction; toll-like receptor 3 signaling pathway; tumor necrosis factor-mediated signaling pathway
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Product Notes
The Tnf tnf (Catalog #AAA1753257) is an Antibody produced from Rabbit and is intended for research purposes only. The product is available for immediate purchase. The Anti-TNF alpha/Tnf Antibody reacts with Mouse, Rat and may cross-react with other species as described in the data sheet. AAA Biotech's TNF alpha/Tnf can be used in a range of immunoassay formats including, but not limited to, Western Blot (WB), Immunocytochemistry (ICC), Immunofluorescence (IF), Flow Cytometry (FC/FACS/FCM), Direct ELISA (EIA). WB: 0.25-0.5ug/ml|Mouse, Rat| ICC/IF: 5ug/ml|Mouse| FC/FACS/FCM: 1-3ug/1x106 cells|Mouse| Direct ELISA: 0.1-0.5ug/ml|Mouse|. Researchers should empirically determine the suitability of the Tnf tnf for an application not listed in the data sheet. Researchers commonly develop new applications and it is an integral, important part of the investigative research process. It is sometimes possible for the material contained within the vial of "TNF alpha/Tnf, Polyclonal Antibody" to become dispersed throughout the inside of the vial, particularly around the seal of said vial, during shipment and storage. We always suggest centrifuging these vials to consolidate all of the liquid away from the lid and to the bottom of the vial prior to opening. Please be advised that certain products may require dry ice for shipping and that, if this is the case, an additional dry ice fee may also be required.Precautions
All products in the AAA Biotech catalog are strictly for research-use only, and are absolutely not suitable for use in any sort of medical, therapeutic, prophylactic, in-vivo, or diagnostic capacity. By purchasing a product from AAA Biotech, you are explicitly certifying that said products will be properly tested and used in line with industry standard. AAA Biotech and its authorized distribution partners reserve the right to refuse to fulfill any order if we have any indication that a purchaser may be intending to use a product outside of our accepted criteria.Disclaimer
Though we do strive to guarantee the information represented in this datasheet, AAA Biotech cannot be held responsible for any oversights or imprecisions. AAA Biotech reserves the right to adjust any aspect of this datasheet at any time and without notice. It is the responsibility of the customer to inform AAA Biotech of any product performance issues observed or experienced within 30 days of receipt of said product. To see additional details on this or any of our other policies, please see our Terms & Conditions page.Item has been added to Shopping Cart
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