Neurofilament Antibody Sampler Kit
cat.: HAK21090
| Product Type: | Antibody Sampler Kit |
|---|---|
| Species reactivity: | Human, Mouse, Rat |
| Applications: | WB, IHC-P, IF-Tissue |
| Form: | Liquid |
| Storage condition: | Store at +4℃ after thawing. Aliquot store at -20℃. Avoid repeated freeze / thaw cycles. |
| Storage buffer: | 1*TBS (pH7.4), 0.05% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide. |
| Uniprot #: | SwissProt: P07197 Human | P08553 Mouse | P12839 Rat | P07196 Human | P08551 Mouse | P19527 Rat | P12036 Human | P19246 Mouse | P16884 Rat |
Images
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Fig1:
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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Fig2:
Western blot analysis of NF-L on different lysates with Rabbit anti-NF-L antibody (HA721538) at 1/1,000 dilution. Lane 1: Mouse brain tissue lysate Lane 2: Rat brain tissue lysate Lane 3: Mouse hippocampus tissue lysate Lane 4: Rat hippocampus tissue lysate Lane 5: Human liver tissue lysate (negative) Lane 6: Mouse liver tissue lysate (negative) Lane 7: Rat liver tissue lysate (negative) Lysates/proteins at 20 µg/Lane. Predicted band size: 62 kDa Observed band size: 68 kDa Exposure time: 3 minutes; 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 (HA721538) at 1/1,000 dilution was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:100,000 dilution was used for 1 hour at room temperature. |
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Fig3:
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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Fig4:
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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Fig5:
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). |
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Fig6:
Immunocytochemistry analysis of mouse glial cells labeling NF-L with Rabbit anti-NF-L antibody (HA721538) at 1/1,000 dilution. Cells were fixed with 4% PFA (15 min), permeabilized with 0.25% TritonX-100 for 15 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1%PBS-Tween for 1h. The cells were then incubated overnight at 4℃ with Rabbit anti-NF-L antibody (HA721538) at at 1/1,000 dilution. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI. |
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Fig7:
Immunofluorescence analysis of frozen mouse cerebellum tissue with Rabbit anti-NF-L antibody (HA721538) 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 (HA721538, 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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Fig8:
Immunohistochemical analysis of paraffin-embedded rat cerebellum tissue with Rabbit anti-NF-L antibody (HA721538) 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 (HA721538) 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. |
Note: All products are “FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE”.