alpha Internexin Recombinant Rabbit Monoclonal Antibody [JU25-03]
cat.: ET1706-23
| Product Type: | Recombinant Rabbit monoclonal IgG, primary antibodies |
|---|---|
| Species reactivity: | Human, Mouse, Rat |
| Applications: | WB, IF-Cell, IF-Tissue, IHC-P, FC, IP |
| Clonality: | Monoclonal |
| Clone number: | JU25-03 |
| 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: 55 kDa |
| Isotype: | IgG |
| Immunogen: | Synthetic peptide within Human alpha Internexin aa 394-443 / 499. |
| Positive control: | SH-SY5Y cell lysate, Human brain tissue lysate, Mouse brain tissue lysate, Rat brain tissue lysate, SH-SY5Y, human brain tissue, mouse brain tissue, rat brain tissue. |
| Subcellular location: | Intermediate filament cytoskeleton, neurofilament, extracellular space, cytoplasmic ribonucleoprotein granule, postsynapse, Schaffer collateral - CA1 synapse. |
| Recommended Dilutions:
WB IF-Cell IF-Tissue IHC-P FC IP |
1:2,000-1:5,000 1:100 1:50-1:200 1:50-1:200 1:1,000 1-2μg/sample |
| Uniprot #: | SwissProt: Q16352 Human | P46660 Mouse | P23565 Rat |
| Alternative names: | 66 kDa neurofilament protein AINX_HUMAN Alpha Inx Alpha-internexin Alpha-Inx INA Internexin neuronal intermediate filament protein alpha MGC12702 NEF 5 NEF5 Neurofilament 5 (66kD) Neurofilament 5 Neurofilament 66 Neurofilament 66 tax binding protein Neurofilament-66 NF 66 NF-66 NF66 TXBP 1 TXBP1 |
Images
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Fig1:
Western blot analysis of alpha Internexin on different lysates with Rabbit anti-alpha Internexin antibody (ET1706-23) at 1/2,000 dilution. Lane 1: SH-SY5Y cell lysate Lane 2: Human brain tissue lysate Lane 3: Mouse brain tissue lysate Lane 4: Rat brain tissue lysate Lane 5: Mouse pancreas tissue lysate (negative) Lysates/proteins at 20 µg/Lane. Predicted band size: 55 kDa Observed band size: 60 kDa Exposure time: 6 seconds; 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 (ET1706-23) 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:
Immunocytochemistry analysis of SH-SY5Y cells labeling alpha Internexin with Rabbit anti-alpha Internexin antibody (ET1706-23) at 1/100 dilution. Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 15 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-alpha Internexin antibody (ET1706-23) at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. 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. Beta tubulin (HA601187, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution. |
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Fig3:
Immunohistochemical analysis of paraffin-embedded human brain tissue with Rabbit anti-alpha Internexin antibody (ET1706-23) 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 (ET1706-23) 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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Fig4:
Immunohistochemical analysis of paraffin-embedded mouse brain tissue with Rabbit anti-alpha Internexin antibody (ET1706-23) 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 (ET1706-23) 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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Fig5:
Immunohistochemical analysis of paraffin-embedded rat brain tissue with Rabbit anti-alpha Internexin antibody (ET1706-23) 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 (ET1706-23) 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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Fig6:
Immunofluorescence analysis of paraffin-embedded rat brain tissue labeling alpha Internexin with Rabbit anti-alpha Internexin antibody (ET1706-23) at 1/50 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 (ET1706-23, green) at 1/50 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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Fig7:
Flow cytometric analysis of SH-SY5Y cells labeling alpha Internexin. Cells were fixed and permeabilized. Then stained with the primary antibody (ET1706-23, 1/1,000) (red) compared with Rabbit IgG Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a iFluor™ 488 conjugate-Goat anti-Rabbit IgG Secondary antibody (HA1121) at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black). |
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Fig8:
alpha Internexin was immunoprecipitated from 0.2 mg SH-SY5Y cell lysate with ET1706-23 at 2 µg/10 µl beads. Western blot was performed from the immunoprecipitate using ET1706-23 at 1/2,000 dilution. Anti-Rabbit IgG for IP Nano-secondary antibody (NBI01H) at 1/5,000 dilution was used for 1 hour at room temperature. Lane 1: SH-SY5Y cell lysate (input) Lane 2: ET1706-23 IP in SH-SY5Y cell lysate Lane 3: Rabbit IgG instead of ET1706-23 in SH-SY5Y cell lysate Blocking/Dilution buffer: 5% NFDM/TBST Exposure time: 12 seconds; ECL: K1801 |
Note: All products are “FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE”.