WB
Western blot analysis using Anti-TNFa-Human antibody against Recombinant TNFa-Human protein. 15% SDS-PAGE gel.Sample loading: 10ng /lane. Transfer the proteins onto a PVDF membrane (OM790002), and block it with TBST (OM750016) plus skimmed milk powder(OM750084) for one hour. Dilute the primary antibody with the antibody diluent (OM750012) at a ratio of 1:5000, and incubate it overnight at 4°C. Wash the membrane three times with TBST (OM750016), 5 minutes each time. At room temperature, dilute the secondary antibody, Goat Anti-Rabbit IgG(H&L)-HRP (OM643487), at a ratio of 1:10000 and incubate for one hour. Wash the membrane three times with TBST (OM750016) again, 5 minutes each time. Use ECL (OM625701) for luminescence.staining time: 30S.WB
Western blot analysis using TNFa antibody against Hela(1),Nih3t3(2) cell . 12% SDS-PAGE gel.Sample loading: 20μg /lane. Transfer the proteins onto a PVDF membrane (OM790003), and block it with TBST (OM750016) plus skimmed milk powder(OM750084) for one hour. Dilute the primary antibody with the antibody diluent (OM750012) at a ratio of 1:1000, and incubate it overnight at 4°C. Wash the membrane three times with TBST (OM750016), 5 minutes each time. At room temperature, dilute the secondary antibody, Goat Anti-Rabbit IgG(H&L)-HRP (OM643487), at a ratio of 1:20000 and incubate for one hour. Wash the membrane three times with TBST (OM750016) again, 5 minutes each time. Use ECL (OM625701) for luminescence.staining time: 60S.ELISA
Microtiter plate wells were coated with 100 ng/well coating antigen and incubated with serial dilutions of the antibody. Following HRP-conjugated secondary antibody and substrate development, OD450 was measured. The absorbance decreased progressively with increasing antibody dilution, yielding a sigmoidal titration curve that demonstrates the high antigen-binding activity of the polyclonal antibody.ICC/IF
Immunofluorescence analysis of A549 cells using TNFa antibody (green). Blue: DAPI fluorescent DNA dye. Red: Actin filaments have been labeled with Omnimabs® 594-Phalloidin.Cells are fixed in 4% paraformaldehyde at room temperature for 15 minutes. Then, they are permeabilized with a PBS (OM750003) solution containing 0.1% Triton X-100(OM750021) at room temperature for 15 minutes. Subsequently, the cells are blocked with 10% non - immune goat serum(OM760028) at room temperature for 1 hour.The cells are incubated overnight at 4°C with the primary antibody diluted 1:100 in PBS. The secondary antibody, Omnimabs® 488 Goat Ant-Rabbit IgG(H&L) (Green,OM643486), is diluted at a ratio of 1:400 and incubated with the cells for 1 hour.Nuclear DNA is labeled with DAPI (Blue,OM643160). F-actin is stained with Omnimabs® 594-Phalloidin (Red,OM750007) diluted 1:100 for 30 minutes.| Product Name | Anti-TNFa antibody |
|---|---|
| Antibody Type | Primary Antibodies |
| Immunogen | Recombinant Human TNF protein |
| Clonality | polyclonal |
|---|---|
| Isotype | IgG |
| Host Species | Rabbit |
| Tested Applications | ELISAICC/IFWB |
| ELISA:1:1000-1:10000 WB:1:1000-1:2000 ICC/IF:1:100-1:500 |
|
| Species Reactivity | HumanMouse |
| Concentration | 1mg/ml |
| Purification | Affinity purified |
| Gene Symbol | TNF |
|---|---|
| Gene Synonyms | cTNF DIF IMD127 RATTNF TNF-a TNF-alpha TNFA TNFalpha Tnfsf1a TNFSF2 TNLG1F |
| Gene Full Name | tumor necrosis factor |
| Gene Summary | 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, psoriasis, rheumatoid arthritis ankylosing spondylitis, tuberculosis, autosomal dominant polycystic kidney disease, and cancer. Mutations in this gene affect susceptibility to cerebral malaria, septic shock, and Alzheimer disease. Knockout studies in mice also suggested the neuroprotective function of this cytokine. [provided by RefSeq, Aug 2020] |
| Alternative Names | APC1 protein cachectin cell apoptosis-related protein cytokine DADB-70P7.1 pro-tumor necrosis factor TNF alpha TNF superfamily TNF superfamily member 2 TNF α TNF macrophage-derived TNF monocyte-derived TNF-a TNF-alpha TNFα tumor necrosis factor tumor necrosis factor (TNF superfamily member 2) tumor necrosis factor alpha tumor necrosis factor ligand 1F tumor necrosis factor ligand superfamily member 2 tumor necrosis factor precursor tumor necrosis factor-alpha tumor necrotic factor alpha tumour necrosis factor |
| Molecular Weight(MW) | 26kDa（observed：35、37kDa) |
| Cellular Localization | Secreted, membrane, cytoplasm. |
| SwissProt ID | P01375 |
|---|
WB
Western blot analysis using Anti-TNFa-Human antibody against Recombinant TNFa-Human protein. 15% SDS-PAGE gel.Sample loading: 10ng /lane. Transfer the proteins onto a PVDF membrane (OM790002), and block it with TBST (OM750016) plus skimmed milk powder(OM750084) for one hour. Dilute the primary antibody with the antibody diluent (OM750012) at a ratio of 1:5000, and incubate it overnight at 4°C. Wash the membrane three times with TBST (OM750016), 5 minutes each time. At room temperature, dilute the secondary antibody, Goat Anti-Rabbit IgG(H&L)-HRP (OM643487), at a ratio of 1:10000 and incubate for one hour. Wash the membrane three times with TBST (OM750016) again, 5 minutes each time. Use ECL (OM625701) for luminescence.staining time: 30S.
WB
Western blot analysis using TNFa antibody against Hela(1),Nih3t3(2) cell . 12% SDS-PAGE gel.Sample loading: 20μg /lane. Transfer the proteins onto a PVDF membrane (OM790003), and block it with TBST (OM750016) plus skimmed milk powder(OM750084) for one hour. Dilute the primary antibody with the antibody diluent (OM750012) at a ratio of 1:1000, and incubate it overnight at 4°C. Wash the membrane three times with TBST (OM750016), 5 minutes each time. At room temperature, dilute the secondary antibody, Goat Anti-Rabbit IgG(H&L)-HRP (OM643487), at a ratio of 1:20000 and incubate for one hour. Wash the membrane three times with TBST (OM750016) again, 5 minutes each time. Use ECL (OM625701) for luminescence.staining time: 60S.
ELISA
Microtiter plate wells were coated with 100 ng/well coating antigen and incubated with serial dilutions of the antibody. Following HRP-conjugated secondary antibody and substrate development, OD450 was measured. The absorbance decreased progressively with increasing antibody dilution, yielding a sigmoidal titration curve that demonstrates the high antigen-binding activity of the polyclonal antibody.
ICC/IF
Immunofluorescence analysis of A549 cells using TNFa antibody (green). Blue: DAPI fluorescent DNA dye. Red: Actin filaments have been labeled with Omnimabs® 594-Phalloidin.Cells are fixed in 4% paraformaldehyde at room temperature for 15 minutes. Then, they are permeabilized with a PBS (OM750003) solution containing 0.1% Triton X-100(OM750021) at room temperature for 15 minutes. Subsequently, the cells are blocked with 10% non - immune goat serum(OM760028) at room temperature for 1 hour.The cells are incubated overnight at 4°C with the primary antibody diluted 1:100 in PBS. The secondary antibody, Omnimabs® 488 Goat Ant-Rabbit IgG(H&L) (Green,OM643486), is diluted at a ratio of 1:400 and incubated with the cells for 1 hour.Nuclear DNA is labeled with DAPI (Blue,OM643160). F-actin is stained with Omnimabs® 594-Phalloidin (Red,OM750007) diluted 1:100 for 30 minutes.| Application Notes | ELISA:1:1000-1:10000 WB:1:1000-1:2000 ICC/IF:1:100-1:500 |
|---|
| Form | Liquid |
|---|---|
| Storage Instructions | Shipped at 4°C. Store at +4°C short term (1-2 weeks). Store at -20°C long term. Avoid freeze / thaw cycle. |
| Storage Buffer | Purified antibody in PBS with 0.05% sodium azide,40% Glycerol. |
