Images
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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.
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☑ Knockdown (KD)
Western blot analysis of Huntingtin on different lysates with Rabbit anti-Huntingtin antibody (ET7107-60) at 1/10,000 dilution.
Lane 1: HAP1-parental cell lysate
Lane 2: HAP1-Huntingtin KD cell lysate
Lysates/proteins at 10 µg/Lane.
Predicted band size: 348 kDa
Observed band size: 348 kDa
Exposure time: 100 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 (ET7107-60) at 1/10,000 dilution was used in K1803 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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Application: IHC-Fr
Species: Mouse
Site: Cerebral cortex
Sample: Frozen section
Antibody concentration: 1/200
Antigen retrieval: Not required
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Application: IF-tissue
Species: Mouse
Site: Cerebral cortex
Sample: Paraffin-embedded section
Antibody concentration: 1/200
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Immunohistochemical analysis of paraffin-embedded human brain tissue with Rabbit anti-Huntingtin antibody (ET7107-60) 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 (ET7107-60) 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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Immunohistochemical analysis of paraffin-embedded mouse cerebellum tissue with Rabbit anti-Huntingtin antibody (ET7107-60) 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 (ET7107-60) 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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Immunohistochemical analysis of paraffin-embedded rat cerebellum tissue with Rabbit anti-Huntingtin antibody (ET7107-60) 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 (ET7107-60) 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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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.
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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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