Anti-Alpha-synuclein, isoform 98 and 112 antibody [EPR25115-65] - BSA and Azide free

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Alpha-synuclein, isoform 98 and 112 antibody [EPR25115-65] - BSA and Azide free (AB288163)

This data was developed using ab288156, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin-embedded (A) HEK-293T cells transfected with a SNCA112 expression vector containing a his tag. (B) HEK-293T cells transfected with a SNCA98 expression vector containing a his tag. (C) HEK-293T cells transfected with a SNCA140 expression vector containing a his tag. (D) HEK-293T cells transfected with a SNCA126 expression vector containing a his tag labelling Alpha-synuclein, (isoform 98 and 112) with ab288156 at 1/1000 (5.24 ug/ml) followed by a ready to use LeicaDS9800 (Bond™, Polymer Refine Detection).

Posiitive staining on (A) HEK-293T cells transfected with a SNCA112 and (B) HEK-293T cells transfected with a SNCA98.

No staining on (C) HEK-293T cells transfected with a SNCA140 and (D) HEK-293T cells transfected with a SNCA126.

The section was incubated with ab288156 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).

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

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-Alpha-synuclein, isoform 98 and 112 antibody [EPR25115-65] - BSA and Azide free (AB288163)

This data was developed using ab288156, the same antibody clone in a different buffer formulation.

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HEK-293T cells labelling Alpha-synuclein, isoform 98 and 112 with ab288156 at 1/100 (5.24 ug/ml) 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 cytoplasmic and nuclear staining in HEK-293T cells transfected with a human SNCA112 expression vector and a human SNCA98 expression vector containing a myc tag.

No staining in HEK-293T cells transfected with a human SNCA126 expression vector or a human SNCA140 expression vector containing a myc tag.

Myc-Tag Mouse mAb (Alexa Fluor^® 647) was used to counterstain myc tag at 1/100 0.38ug/ml 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 2 ug/ml dilution.

• Flow Cyt (Intra)

Lab

Flow Cytometry (Intracellular) - Anti-Alpha-synuclein, isoform 98 and 112 antibody [EPR25115-65] - BSA and Azide free (AB288163)

Intracellular Flow Cytometry analysis of HEK-293T (Human embryonic kidney epithelial cell) cells transfected with a human SNCA126 expression vector containing a his tag (Left) / HEK-293T cells transfected with a human SNCA98 expression vector containing a his tag (Right) labeling Alpha-synuclein, isoform 98 and 112 with ab288156 at 1/500 dilution (Left and Right). Cells were fixed with 4% Paraformaldehyde and permeabilised with 90% Methanol. A Goat anti rabbit IgG (Alexa Fluor^® 488, ab150081) secondary antibody was used at 1/2000 dilution.

This data was developed using ab288156, the same antibody clone in a different buffer formulation.

• Flow Cyt (Intra)

Lab

Flow Cytometry (Intracellular) - Anti-Alpha-synuclein, isoform 98 and 112 antibody [EPR25115-65] - BSA and Azide free (AB288163)

Intracellular Flow Cytometry analysis of HEK-293T (Human embryonic kidney epithelial cell) cells transfected with a human SNCA140 expression vector containing a his tag (Left) / HEK-293T cells transfected with a human SNCA112 expression vector containing a his tag (Right) labeling Alpha-synuclein, isoform 98 and 112 with ab288156 at 1/500 dilution (Left and Right). Cells were fixed with 4% Paraformaldehyde and permeabilised with 90% Methanol. A Goat anti rabbit IgG (Alexa Fluor^® 488, ab150081) secondary antibody was used at 1/2000 dilution.

This data was developed using ab288156, the same antibody clone in a different buffer formulation.

• WB

Lab

Western blot - Anti-Alpha-synuclein, isoform 98 and 112 antibody [EPR25115-65] - BSA and Azide free (AB288163)

This data was developed using ab288156, the same antibody clone in a different buffer formulation.

This antibody can recognize human SNCA112 and human SNCA98, but not human SNCA140 and human SNCA126.

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

Exposure time : 3 minutes.

All lanes:

Western blot - Anti-Alpha-synuclein, isoform 98 and 112 antibody [EPR25115-65] (<a href='/en-us/products/primary-antibodies/alpha-synuclein-isoform-98-and-112-antibody-epr25115-65-ab288156'>ab288156</a>) at 1/1000 dilution

Lane 1:

HEK-293T cells (human embryonic kidney epithelial cell) transfected with a human SNCA140 expression vector containing a his tag, whole cell lysate at 10 µg

Lane 2:

HEK-293T cells transfected with a human SNCA126 expression vector containing a his tag, whole cell lysate at 10 µg

Lane 3:

HEK-293T cells transfected with a human SNCA112 expression vector containing a his tag, whole cell lysate at 10 µg

Lane 4:

HEK-293T cells transfected with a human SNCA98 expression vector containing a his tag, whole cell lysate at 10 µg

Secondary

All lanes:

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

Predicted band size: 14 kDa

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