Product Type: | Recombinant Rabbit monoclonal IgG, primary antibodies |
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Species reactivity: | Human, Mouse, Rat |
Applications: | WB, IF-Cell, IF-Tissue, IHC-P, IP |
Clonality: | Monoclonal |
Clone number: | JM11-20 |
Form: | Liquid |
Storage condition: | Store at +4℃ after thawing. Aliquot store at -20℃ or -80℃. Avoid repeated freeze / thaw cycles. |
Storage buffer: | 1*TBS (pH7.4), 0.05% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide. |
Concentration: | 1ug/ul |
Purification: | Protein A affinity purified. |
Molecular weight: | Predicted band size: 102 kDa |
Isotype: | IgG |
Immunogen: | Synthetic peptide within Human NEFM aa 651-692 / 916. |
Positive control: | 293T cell lysates, N2A, PC-12, SH-SY5Y, mouse brain tissue, rat brain tissue, human brain tissue. |
Subcellular location: | Cell projection, Cytoplasm, Cytoskeleton, Intermediate filament. |
Recommended Dilutions:
WB IF-Cell IF-Tissue IHC-P IP |
1:1,000-1:2,000 1:100-1:500 1:100-1:500 1:50-1:200 Use at an assay dependent concentration. |
Uniprot #: | SwissProt: P07197 Human | P08553 Mouse | P12839 Rat |
Alternative names: | 150kDa medium NEF3 NEFM Neurofilament 3 NF160 NFM |
Fig1: Western blot analysis of NEFM on 293T cell lysates. Proteins were transferred to a PVDF membrane and blocked with 5% BSA in PBS for 1 hour at room temperature. The primary antibody (ET1703-57, 1/500) was used in 5% BSA at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:200,000 dilution was used for 1 hour at room temperature. | |
Fig2: ICC staining of NEFM in N2A cells (red). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 1% Blocker BSA for 15 minutes at room temperature. Cells were probed with the primary antibody (ET1703-57, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor®594 Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue). | |
Fig3: ICC staining of NEFM in PC-12 cells (red). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 1% Blocker BSA for 15 minutes at room temperature. Cells were probed with the primary antibody (ET1703-57, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor®594 Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue). | |
Fig4: ICC staining of NEFM in SH-SY5Y cells (red). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 1% Blocker BSA for 15 minutes at room temperature. Cells were probed with the primary antibody (ET1703-57, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor®594 Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue). |
Fig5: Immunohistochemical analysis of paraffin-embedded mouse brain tissue using anti-NEFM antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) for 20 minutes.The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1703-57, 1/50) for 30 minutes at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX. | |
Fig6: Immunohistochemical analysis of paraffin-embedded rat brain tissue using anti-NEFM antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) for 20 minutes.The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1703-57, 1/50) for 30 minutes at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX. | |
Fig7:
Immunohistochemical analysis of paraffin-embedded human brain tissue with Rabbit anti-NEFM antibody (ET1703-57) at 1/400 dilution. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1703-57) at 1/400 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX. |