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: 55 kDa
Isotype: IgG
Immunogen: Synthetic peptide within Human alpha Internexin aa 394-443 / 499.
Positive control: A549 cell lysate, mouse brain tissue lysate, PC-12 cell lysate, PC-12, rat brain tissue, mouse cerebellum tissue, N2A.
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
1:1,000-1:5,000
1:50-1:200
1:50-1:200
1:50-1:200
1:50-1:100
Use at an assay dependent concentration.
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
ET1706-23_1.jpg Fig1: Western blot analysis of alpha Internexin on different lysates with Rabbit anti-alpha Internexin antibody (ET1706-23) at 1/500 dilution.

Lane 1: A549 cell lysate
Lane 2: mouse brain tissue lysate(20 µg/Lane)
Lane 3: PC-12 cell lysate

Lysates/proteins at 10 µg/Lane.

Predicted band size: 55 kDa
Observed band size: 55 kDa

Exposure time: 2 minutes;

10% 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 1/500 dilution was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 200,000 dilution was used for 1 hour at room temperature.
ET1706-23_2.jpg Fig2: Immunocytochemistry analysis of PC-12 cells labeling alpha Internexin with Rabbit anti-alpha Internexin antibody (ET1706-23) at 1/50 dilution.

Cells were fixed in 4% paraformaldehyde for 10 minutes at 37 ℃, permeabilized with 0.05% Triton X-100 in PBS for 20 minutes, and then blocked with 2% negative goat serum for 30 minutes at room temperature. Cells were then incubated with Rabbit anti-alpha Internexin antibody (ET1706-23) at 1/50 dilution in 2% negative goat serum overnight at 4 ℃. Alexa Fluor®488 conjugate-Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. Nuclear DNA was labelled in blue with DAPI.
ET1706-23_3.jpg Fig3: Immunohistochemical analysis of paraffin-embedded rat brain tissue 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 8.0-8.4) 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 1/50 dilution for 0.5 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.
ET1706-23_4.jpg Fig4: Immunohistochemical analysis of paraffin-embedded mouse cerebellum tissue 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 8.0-8.4) 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 1/50 dilution for 0.5 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.
ET1706-23_5.jpg Fig5: Flow cytometric analysis of alpha Internexin was done on N2A cells. The cells were fixed, permeabilized and stained with the primary antibody (ET1706-23, 1/50) (red). After incubation of the primary antibody at room temperature for an hour, the cells were stained with a Alexa Fluor 488-conjugated Goat anti-Rabbit IgG Secondary antibody at 1/1000 dilution for 30 minutes.Unlabelled sample was used as a control (cells without incubation with primary antibody; black).
ET1706-23_6.jpg 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).
Note: All products are “FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE”.