Neurofilament heavy polypeptide Recombinant Rabbit Monoclonal Antibody [JF99-07]
cat.: ET1702-72
| Product Type: | Recombinant Rabbit monoclonal IgG, primary antibodies |
|---|---|
| Species reactivity: | Human, Mouse, Rat |
| Applications: | WB, IHC-P, IHC-Fr, IF-Tissue |
| Clonality: | Monoclonal |
| Clone number: | JF99-07 |
| 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: 200 kDa |
| Isotype: | IgG |
| Immunogen: | Synthetic peptide within human NEFH aa 690-740. |
| Positive control: | Mouse brain tissue lysate, Rat brain tissue lysate, Rat hippocampus tissue lysate, human cerebellum tissue, mouse cerebellum tissue, rat cerebellum tissue. |
| Subcellular location: | Cytoplasm. |
| Recommended Dilutions:
WB IHC-P IHC-Fr IF-Tissue |
1:2,000 1:1,000 1:500 1:500 |
| Uniprot #: | SwissProt: P12036 Human | P19246 Mouse | P16884 Rat |
| Alternative names: | 200 kDa neurofilament protein CMT2CC Nefh Neurofilament heavy polypeptide 200kDa Neurofilament heavy polypeptide Neurofilament triplet H protein NF H NF-H NFH NFH_HUMAN |
Images
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Fig1:
Immunofluorescence analysis of frozen mouse cerebellum tissue with Rabbit anti-Neurofilament heavy polypeptide antibody (ET1702-72) at 1/500 dilution. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for about 2 minutes in microwave oven. The tissues were blocked in 10% negative goat serum for 1 hour at room temperature, washed with PBS, and then probed with the primary antibody (ET1702-72, green) at 1/500 dilution overnight at 4 ℃, washed with PBS. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. Nuclei were counterstained with DAPI (blue). |
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Fig2:
Immunofluorescence analysis of frozen mouse liver tissue (negative) with Rabbit anti-Neurofilament heavy polypeptide antibody (ET1702-72) at 1/500 dilution. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for about 2 minutes in microwave oven. The tissues were blocked in 10% negative goat serum for 1 hour at room temperature, washed with PBS, and then probed with the primary antibody (ET1702-72, green) at 1/500 dilution overnight at 4 ℃, washed with PBS. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. Nuclei were counterstained with DAPI (blue). |
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Fig3:
Immunofluorescence analysis of frozen rat cerebellum tissue with Rabbit anti-Neurofilament heavy polypeptide antibody (ET1702-72) at 1/500 dilution. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for about 2 minutes in microwave oven. The tissues were blocked in 10% negative goat serum for 1 hour at room temperature, washed with PBS, and then probed with the primary antibody (ET1702-72, green) at 1/500 dilution overnight at 4 ℃, washed with PBS. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. Nuclei were counterstained with DAPI (blue). |
|
Fig4:
Immunofluorescence analysis of frozen rat liver tissue (negative) with Rabbit anti-Neurofilament heavy polypeptide antibody (ET1702-72) at 1/500 dilution. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for about 2 minutes in microwave oven. The tissues were blocked in 10% negative goat serum for 1 hour at room temperature, washed with PBS, and then probed with the primary antibody (ET1702-72, green) at 1/500 dilution overnight at 4 ℃, washed with PBS. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. Nuclei were counterstained with DAPI (blue). |
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Fig5:
Western blot analysis of Neurofilament heavy polypeptide on different lysates with Rabbit anti-Neurofilament heavy polypeptide antibody (ET1702-72) at 1/2,000 dilution. Lane 1: Mouse brain tissue lysate (20 µg/Lane) Lane 2: Rat brain tissue lysate (20 µg/Lane) Lane 3: Rat hippocampus tissue lysate (20 µg/Lane) Predicted band size: 200 kDa Observed band size: 250 kDa Exposure time: 3 minutes; ECL: K1801; 4-20% SDS-PAGE gel. Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (ET1702-72) at 1/2,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature. |
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Fig6:
Immunohistochemical analysis of paraffin-embedded human cerebellum tissue with Rabbit anti-Neurofilament heavy polypeptide antibody (ET1702-72) at 1/200 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 (ET1702-72) at 1/200 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. |
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Fig7:
Immunohistochemical analysis of paraffin-embedded mouse cerebellum tissue with Rabbit anti-Neurofilament heavy polypeptide antibody (ET1702-72) at 1/1,000 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 (ET1702-72) at 1/1,000 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. |
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Fig8:
Immunohistochemical analysis of paraffin-embedded rat cerebellum tissue with Rabbit anti-Neurofilament heavy polypeptide antibody (ET1702-72) at 1/1,000 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 (ET1702-72) at 1/1,000 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. |
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Fig9:
Immunofluorescence analysis of paraffin-embedded mouse cerebellum tissue labeling Neurofilament heavy polypeptide with Rabbit anti-Neurofilament heavy polypeptide antibody (ET1702-72) at 1/500 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 10% negative goat serum for 1 hour at room temperature, washed with PBS, and then probed with the primary antibody (ET1702-72, green) at 1/500 dilution overnight at 4 ℃, washed with PBS. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. Nuclei were counterstained with DAPI (blue). |
Note: All products are “FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE”.