Huntingtin Recombinant Rabbit Monoclonal Antibody [JB89-34]
cat.: ET7107-60
Product Type: Recombinant Rabbit monoclonal IgG, primary antibodies
Species reactivity: Human, Mouse, Rat
Applications: WB, IHC-P, FC, IHC-Fr, IF-Cell, IF-Tissue
Clonality: Monoclonal
Clone number: JB89-34
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: 348 kDa
Isotype: IgG
Immunogen: Synthetic peptide within N-terminal Human Huntingtin .
Positive control: Neuro-2a cell lysate, PC-12 cell lysate, mouse brain tissue lysate, rat brain tissue lysate, human colon carcinoma tissue, human breast tissue, mouse epididymis tissue, rat brain tissue, SH-SY5Y.
Subcellular location: Cytoplasm. Nucleus.
Recommended Dilutions:
  WB
  IHC-P
  FC
  IHC-Fr
  IF-Cell
  IF-Tissue
1:5,000
1:50-1:200
1:50-1:100
1:100
1:100
1:50
Uniprot #: SwissProt: P42858 Human | P42859 Mouse | P51111 Rat
Alternative names: AI256365 C430023I11Rik HD HD protein HD_HUMAN HDH HTT Huntingtin HUNTINGTON CHOREA Huntington disease protein Huntington's disease protein homolog IT 15 IT15 OTTMUSP00000026909 ZHD
Images
ET7107-60_1.jpg Fig1: Western blot analysis of Huntingtin on different lysates with Rabbit anti-Huntingtin antibody (ET7107-60) at 1/5,000 dilution.

Lane 1: Neuro-2a cell lysate (15 µg/Lane)
Lane 2: PC-12 cell lysate (15 µg/Lane)
Lane 3: Mouse brain tissue lysate (20 µg/Lane)
Lane 4: Rat brain tissue lysate (20 µg/Lane)

Predicted band size: 348 kDa
Observed band size: 348 kDa

Exposure time: 2 minutes 17 seconds;

3-8% 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 (ET7107-60) at 1/5,000 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.
ET7107-60_2.jpg Fig2: Immunohistochemical analysis of paraffin-embedded human colon carcinoma tissue using anti-Huntingtin antibody. 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 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET7107-60, 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.
ET7107-60_3.jpg Fig3: Immunohistochemical analysis of paraffin-embedded human breast tissue using anti-Huntingtin antibody. 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 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET7107-60, 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.
ET7107-60_4.jpg Fig4: Immunohistochemical analysis of paraffin-embedded mouse epididymis tissue using anti-Huntingtin antibody. 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 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET7107-60, 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.
ET7107-60_5.jpg Fig5: Immunohistochemical analysis of paraffin-embedded rat brain tissue using anti-Huntingtin antibody. 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 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET7107-60, 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.
ET7107-60_6.jpg Fig6: Immunofluorescence analysis of frozen mouse hippocampus tissue labeling Huntingtin with Rabbit anti-Huntingtin antibody (ET7107-60).
The tissues were blocked in 3% BSA for 30 minutes at room temperature, washed with PBS, and then probed with the primary antibody (ET7107-60, green) at 1/100 dilution overnight at 4℃, washed with PBS. Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) was used as the secondary antibody at 1/200 dilution. Nuclei were counterstained with DAPI (blue). Image acquisition was performed with KFBIO KF-FL-400 Scanner.
ET7107-60_7.jpg Fig7: Immunofluorescence analysis of frozen mouse cerebral cortex tissue labeling Huntingtin with Rabbit anti-Huntingtin antibody (ET7107-60).
The tissues were blocked in 3% BSA for 30 minutes at room temperature, washed with PBS, and then probed with the primary antibody (ET7107-60, green) at 1/100 dilution overnight at 4℃, washed with PBS. Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) was used as the secondary antibody at 1/200 dilution. Nuclei were counterstained with DAPI (blue). Image acquisition was performed with KFBIO KF-FL-400 Scanner.
ET7107-60_8.jpg Fig8: Immunocytochemistry analysis of SH-SY5Y cells labeling Huntingtin with Rabbit anti-Huntingtin antibody (ET7107-60) at 1/100 dilution.

Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 5 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-Huntingtin antibody (ET7107-60) 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 (M1305-2, 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.
ET7107-60_9.jpg Fig9: Immunofluorescence analysis of paraffin-embedded mouse brain tissue labeling Huntingtin with Rabbit anti-Huntingtin antibody (ET7107-60) 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 (ET7107-60, 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).
ET7107-60_10.jpg Fig10: Flow cytometric analysis of Huntingtin was done on SH-SY5Y cells. The cells were fixed, permeabilized and stained with the primary antibody (ET7107-60, 1/50) (red). After incubation of the primary antibody at room temperature for an hour, the cells were stained with a Alexa Fluor®488 conjugate-Goat anti-Rabbit IgG Secondary antibody at 1/1,000 dilution for 30 minutes.Unlabelled sample was used as a control (cells without incubation with primary antibody; black).
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