Anti-alpha + beta Synuclein antibody [EP1646Y]

• Flow Cyt (Intra)

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Flow Cytometry (Intracellular) - Anti-alpha + beta Synuclein antibody [EP1646Y] (AB51252)

Overlay histogram showing U87-MG cells fixed in 4% PFA and stained with purified ab51252 at a dilution of 1 in 30 (red line). The secondary antibody used was FITC goat anti-rabbit at a dilution of 1 in 500. Rabbit monoclonal IgG was used as an isotype control (black line) and cells incubated in the absence of both primary and secondary antibody were used as a negative control (blue line).

• Flow Cyt (Intra)

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Flow Cytometry (Intracellular) - Anti-alpha + beta Synuclein antibody [EP1646Y] (AB51252)

Overlay histogram showing SH-SY5Y cells stained with unpurifiedab51252 (red line). The cells were fixed with 4% paraformaldehyde (10 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (unpurified ab51252, 1/1000 dilution) for 30 min at 22°C. The secondary antibody used was Alexa Fluorr^® 488 goat anti-rabbit IgG (H+L) (ab150077) at 1/2000 dilution for 30 min at 22°C. Isotype control antibody (black line) was rabbit IgG (monoclonal) (0.1μg/1x10⁶ cells) used under the same conditions. Unlabelled sample (blue line) was also used as a control. Acquisition of >5,000 events were collected using a 20mW Argon ion laser (488nm) and 525/30 bandpass filter.

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-alpha + beta Synuclein antibody [EP1646Y] (AB51252)

Immunocytochemistry/Immunofluorescence analysis of PC-12 cells labelling alpha + beta Synuclein with purified ab51252 at 1/100. Cells were fixed with 4% paraformaldehyde and permeabilized with 0.1% Triton X-100. ab150077, an Alexa Fluor^® 488-conjugated goat anti-rabbit IgG (1/1000) was used as the secondary antibody. DAPI (blue) was used as the nuclear counterstain. ab7291, a mouse anti-tubulin (1/1000) and ab150120, an Alexa Fluor^® 594-conjugated goat anti-mouse IgG (1/1000) were also used.

Control 1 : primary antibody (1/100) and secondary antibody, ab150120, an Alexa Fluor^® 594-conjugated goat anti-mouse IgG (1/1000).

Control 2 : ab7291 (1/1000) and secondary antibody, ab150077, an Alexa Fluor^® 488-conjugated goat anti-rabbit IgG (1/1000).

• WB

Lab

Western blot - Anti-alpha + beta Synuclein antibody [EP1646Y] (AB51252)

Blocking and dilution buffer : 5% NFDM /TBST.

All lanes:

Western blot - Anti-alpha + beta Synuclein antibody [EP1646Y] (ab51252) at 1/5000 dilution

All lanes:

Human cerebellum tissue 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 1/20000 dilution

Predicted band size: 14 kDa

Observed band size: 18 kDa

false

• WB

Lab

Western blot - Anti-alpha + beta Synuclein antibody [EP1646Y] (AB51252)

Blocking and dilution buffer : 5% NFDM /TBST.

All lanes:

Western blot - Anti-alpha + beta Synuclein antibody [EP1646Y] (ab51252) at 1/5000 dilution

Lane 1:

Mouse brain tissue lysate at 20 µg

Lane 2:

Rat brain tissue 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 1/20000 dilution

Predicted band size: 14 kDa

Observed band size: 18 kDa

false

• WB

Unknown

Western blot - Anti-alpha + beta Synuclein antibody [EP1646Y] (AB51252)

All lanes:

Western blot - Anti-alpha + beta Synuclein antibody [EP1646Y] (ab51252) at 1/50000 dilution

All lanes:

Human fetal brain tissue lysate at 10 µg

Secondary

All lanes:

HRP-conjugated goat anti-rabbit IgG at 1/2000 dilution

Predicted band size: 14 kDa

Observed band size: 18 kDa

false

• WB

Supplier Data

Western blot - Anti-alpha + beta Synuclein antibody [EP1646Y] (AB51252)

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

This antibody does not cross-react with human beta Synuclein and alpha Synuclein.

In Western blot, Anti-6X His tag® antibody [EPR20547] - ChIP Grade (ab213204) staining at 1/5000 dilution.

In Western blot, Anti-alpha + beta Synuclein antibody [EP1646Y] - (ab51252) staining at 1/1000 dilution.

All lanes:

Western blot - Anti-gamma Synuclein/SNCG antibody [EPR26218-79] (<a href='/en-us/products/primary-antibodies/gamma-synuclein-sncg-antibody-epr26218-79-ab315442'>ab315442</a>) at 1/1000 dilution

Lane 1:

His-tagged human gamma Synuclein recombinant protein at 10 ng

Lane 2:

His-tagged human beta Synuclein recombinant protein at 10 ng

Lane 3:

His-tagged human alpha Synuclein recombinant protein at 10 ng

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

Observed band size: 14 kDa,20 kDa,19 kDa

false

Exposure time: 59s