ELISA
Coat the plate with immunizing antigen.Blue line: Immune serum; Orange line: Pre-immune serum.WB
Western blot analysis using NEFM antibody against Mouse brain tissue(1) . 8% 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.ICC/IF
Immunofluorescence analysis of Hela cells using NEFM 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-NEFM antibody |
|---|---|
| Antibody Type | Primary Antibodies |
| Immunogen | Polypeptide |
| Clonality | Polyclonal |
|---|---|
| Isotype | IgG |
| Host Species | Rabbit |
| Tested Applications | ELISAICC/IFWB |
| WB:1:1000-1:2000 ELISA:1:1000-1:10000 ICC/IF:1:100-1:500 |
|
| Species Reactivity | HumanMouseRat |
| Concentration | 1mg/ml |
| Purification | Protein A |
| Gene Symbol | NEFM |
|---|---|
| Gene Synonyms | NEF3 NF-M NF160 NF165 NFM |
| Gene Full Name | neurofilament medium chain |
| Gene Summary | Neurofilaments are type IV intermediate filament heteropolymers composed of light, medium, and heavy chains. Neurofilaments comprise the axoskeleton and functionally maintain neuronal caliber. They may also play a role in intracellular transport to axons and dendrites. This gene encodes the medium neurofilament protein. This protein is commonly used as a biomarker of neuronal damage. Alternative splicing results in multiple transcript variants encoding distinct isoforms. [provided by RefSeq, Oct 2008] |
| Alternative Names | 160 kDa neurofilament protein medium neurofilament subunit NEFM neurofilament 3 neurofilament 3 (150kDa medium) neurofilament 3 medium neurofilament protein M Neurofilament protein middle polypeptide neurofilament triplet M protein neurofilament triplet M protein 160 KD neurofilament protein NF-M neurofilament medium polypeptide neurofilament medium polypeptide 150kDa neurofilament-3 (150 kD medium) neurofilament-M NF-M NF-M c-terminus NF-M protein NMC |
| Molecular Weight(MW) | 102kDa（observed：160kDa) |
| Source | Rabbit |
| Cellular Localization | Cytoplasm, cytoskeleton;Cell projection, axon;Perikaryon |
| SwissProt ID | P07197 |
|---|
ELISA
Coat the plate with immunizing antigen.Blue line: Immune serum; Orange line: Pre-immune serum.
WB
Western blot analysis using NEFM antibody against Mouse brain tissue(1) . 8% 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.
ICC/IF
Immunofluorescence analysis of Hela cells using NEFM 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 | WB:1:1000-1:2000 ELISA:1:1000-1:10000 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% PC300,40% Glycerol. |
