WB
Western blot analysis using TNFb antibody against C2C12(1),P19(2),Pc-12(3) 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 TNFb 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-TNFb antibody |
|---|---|
| Antibody Type | Primary Antibodies |
| Immunogen | Recombinant Human TNFb 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 | HumanMouseRat |
| Concentration | 1mg/ml |
| Purification | Affinity purified |
| Gene Symbol | LTA |
|---|---|
| Gene Synonyms | hlb382 LT LT-[a] LT-alpha LT[a] LTalpha Ltx TNF-beta TNFB TNFSF1 Tnfsf1b TNLG1E |
| Gene Full Name | lymphotoxin alpha |
| Gene Summary | The encoded protein, a member of the tumor necrosis factor family, is a cytokine produced by lymphocytes. The protein is highly inducible, secreted, and forms heterotrimers with lymphotoxin-beta which anchor lymphotoxin-alpha to the cell surface. This protein also mediates a large variety of inflammatory, immunostimulatory, and antiviral responses, is involved in the formation of secondary lymphoid organs during development and plays a role in apoptosis. Genetic variations in this gene are associated with susceptibility to leprosy type 4, myocardial infarction, non-Hodgkin's lymphoma, and psoriatic arthritis. Alternatively spliced transcript variants have been observed for this gene. [provided by RefSeq, Jul 2012] |
| Alternative Names | DAMA-25N12.13 hypothetical protein lymphotoxin alpha isoform b hypothetical protein lymphotoxin alpha isoform c hypothetical protein lymphotoxin alpha isoform d hypothetical protein lymphotoxin alpha isoform e hypothetical protein lymphotoxin alpha isoform f hypothetical protein lymphotoxin alpha isoform g LT-alpha LTA lymphotoxin A lymphotoxin alpha (TNF superfamily member 1) lymphotoxin precursor lymphotoxin-alpha-like TNF b TNF beta TNF superfamily member 1 TNF β TNF-beta TNFbeta TNFβ tumor necrosis factor beta tumor necrosis factor ligand 1E tumor necrosis factor ligand superfamily member 1 |
| Molecular Weight(MW) | 22kDa（observed：25kDa) |
| Cellular Localization | Membrane, Secreted |
| SwissProt ID | P01374 |
|---|
WB
Western blot analysis using TNFb antibody against C2C12(1),P19(2),Pc-12(3) 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 TNFb 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. |
