Anti-Alpha-synuclein, isoform 98 and 126 antibody [EPR25117-81]

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-Alpha-synuclein, isoform 98 and 126 antibody [EPR25117-81] (AB290644)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeilized 293T (human embryonic kidney epithelial cell) cells lebelling Alpha-synuclein, isoform 98 and 126 with ab290644 at 1/2000 dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor=® 488) preadsorbed antibody at 1/1000 2 ug/ml dilution (Green). Confocal image showing no staining in HEK-293T cells transfected with a human alpha-synuclein isoform SNCA140 or SNCA112 expression vector containing a myc tag is observed. Myc-Tag Mouse mAb (Alexa Fluor^® 647) was used to counterstain tubulin at 1/100 dilution (Red). The nuclear counterstain was DAPI (Blue).

Secondary antibody only control : Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) preadsorbed at 1/1000 dilution.

• Flow Cyt (Intra)

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Flow Cytometry (Intracellular) - Anti-Alpha-synuclein, isoform 98 and 126 antibody [EPR25117-81] (AB290644)

Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized HEK-293T (Human embryonic kidney epithelial cell) transfected with a human Alpha-synuclein, isoform 140 (Left) or Alpha-synuclein, isoform 112 (Right) expression vector containing a myc-His tag cells labelling Alpha-synuclein, isoform 98 and 126 with ab290644 at 1/500 dilution/ Left and Right (Red) compared with a isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody). A Goat Anti-Rabbit IgG (Alexa Fluor^® 488, ab150081) at 1/2000 dilution was used as the secondary antibody. No cross-reactivity with Alpha-synuclein, isoform 140 and isoform 112.

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Alpha-synuclein, isoform 98 and 126 antibody [EPR25117-81] (AB290644)

Immunohistochemical analysis of paraffin-embedded (A) HEK-293T cells tissue labelling Alpha-synuclein, isoform 98 and 126 with ab290644 at 1/100 (4.81 μg/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection) was used. Positive staining on (A) HEK-293T cells transfected with a SNCA98 expression vector containing a his tag and (B) HEK-293T cells transfected with a SNCA126 expression vector containing a his tag. No staining on (C) HEK-293T cells transfected with a SNCA140 expression vector containing a his tag, (D) HEK-293T cells transfected with a SNCA112 expression vector containing a his tag and (E) HEK-293T cells transfected with empty vector containing a his tag. The section was incubated with ab290644 for 30 mins at room temperature.The immunostaining was performed on a Leica Biosystems BOND^® RX instrument. Counterstained with Hematoxylin.

Secondary antibody only control : Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection) was used.

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins

• Flow Cyt (Intra)

Supplier Data

Flow Cytometry (Intracellular) - Anti-Alpha-synuclein, isoform 98 and 126 antibody [EPR25117-81] (AB290644)

Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized HEK-293T (Human embryonic kidney epithelial cell) transfected with a human Alpha-synuclein, isoform 126 (Left) or Alpha-synuclein, isoform 98 (Right) expression vector containing a myc-His tag cells labelling Alpha-synuclein, isoform 98 and 126 with ab290644 at 1/500 dilution/ Left and Right compared with a isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody). A Goat Anti-Rabbit IgG (Alexa Fluor^® 488, ab150081) at 1/2000 dilution was used as the secondary antibody.

• ICC

Supplier Data

Immunocytochemistry - Anti-Alpha-synuclein, isoform 98 and 126 antibody [EPR25117-81] (AB290644)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeilized 293T (human embryonic kidney epithelial cell) cells lebelling Alpha-synuclein, isoform 98 and 126 with ab290644 at 1/2000 dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor^®488) preadsorbed antibody at 1/1000 dilution (Green). Confocal image showing positive staining in HEK-293T cells transfected with a human alpha-synuclein isoform SNCA98 and SNCA126 expression vector containing a myc tag is observed. Myc-Tag Mouse mAb (Alexa Fluor^® 647) was used to counterstain tubulin at 1/100 dilution (Red). The Nuclear counterstain was DAPI (Blue).

Secondary antibody only control : Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) preadsorbed at 1/1000 dilution.

• WB

Unknown

Western blot - Anti-Alpha-synuclein, isoform 98 and 126 antibody [EPR25117-81] (AB290644)

Blocking and diluting buffer and concentration was 5% NFDM/TBST.

This blot was developed using a higher sensitivity ECL substrate.

All lanes:

Western blot - Anti-Alpha-synuclein, isoform 98 and 126 antibody [EPR25117-81] (ab290644) at 1/1000 dilution

Lane 1:

HEK-293T (human embryonic kidney) cells transfected with a human SNCA140 expression vector containing a myc-His tag whole cell lysate at 20 µg

Lane 2:

HEK-293T cells transfected with a human SNCA126 expression vector containing a myc-His tag whole cell lysate at 20 µg

Lane 3:

HEK-293T cells transfected with a human SNCA112 expression vector containing a myc-His tag whole cell lysate at 20 µg

Lane 4:

HEK-293T cells transfected with a human SNCA98 expression vector containing a myc-His tag whole cell lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/100000 dilution

false

Exposure time: 3min