Anti-gamma Synuclein/SNCG antibody [EP1539Y]

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-gamma Synuclein/SNCG antibody [EP1539Y] (AB52633)

Immunofluorescent analysis of 4% Paraformaldehyde fixed, 0.1% Triton X-100 permeabilized T-47D (human ductal breast epithelial tumour epithelial cell) cells and HEK-293 (human embryonic kidney epithelial cell) cells labelling gamma Synuclein/SNCG with the primary antibody at 1/50 (2.06 μg/ml), followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) preadsorbed antibody as secondary at 1/1000 (2 μg/ml) dilution. Confocal image showing nuclear and cytoplasmic staining in T-47D cells (shown in green). ab7291 Anti-alpha Tubulin mouse monoclonal antibody at 1/1000 (1 μg/ml) followed by ab150120 Goat Anti-Mouse IgG H&L (Alexa Fluor^® 594) at 1/1000 (2 μg/ml) was used as a counterstain (shown in magenta). Nuclear DNA was labelled with DAPI (shown in blue).

Negative control : HEK-293 (human embryonic kidney epithelial cell) cells. Negative control 1 was stained with ab52633 at 1/50 followed by ab150120 Goat Anti-Mouse IgG H&L (Alexa Fluor^® 594) at 1/1000 (2 μg/ml). Negative control 2 was stained with ab7291 Anti-alpha Tubulin mouse monoclonal antibody at 1/1000 (1 μg/ml) followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) preadsorbed antibody at 1/1000 (2 μg/ml).

Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-gamma Synuclein/SNCG antibody [EP1539Y] (AB52633)

Immunohistochemical analysis of paraffin-embedded Human adrenal gland tissue labeling gamma Synuclein/SNCG with ab52633 at 1/2000 dilution (0.052 µg/mL).

Positive staining on Human adrenal gland.

The primary antibody was incubated for 10 mins at room temperature.

The immunostaining was performed on a Leica Biosystems BOND^® RX instrument.

Rabbit specific IHC polymer detection kit HRP/DAB (ab209101) was used as the secondary detection system.

Counterstained with Hematoxylin.

Heat mediated antigen retrieval was performed with Citrate buffer (pH 6.0, Epitope Retrieval Solution 1) for 20 mins.

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-gamma Synuclein/SNCG antibody [EP1539Y] (AB52633)

Immunohistochemical analysis of paraffin-embedded Human cerebrum tissue labeling gamma Synuclein/SNCG with ab52633 at 1/2000 dilution (0.052 µg/mL).

Positive staining on human cerebrum.

The primary antibody was incubated for 10 mins at room temperature.

The immunostaining was performed on a Leica Biosystems BOND^® RX instrument.

Rabbit specific IHC polymer detection kit HRP/DAB (ab209101) was used as the secondary detection system.

Counterstained with Hematoxylin.

Heat mediated antigen retrieval was performed with Citrate buffer (pH 6.0, Epitope Retrieval Solution 1) for 20 mins.

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-gamma Synuclein/SNCG antibody [EP1539Y] (AB52633)

Immunohistochemical analysis of paraffin-embedded Human liver tissue labeling gamma Synuclein/SNCG with ab52633 at 1/2000 dilution (0.052 µg/mL).

Low expression : no staining on hepatocytes, but positive staining on vessels in human liver.

The primary antibody was incubated for 10 mins at room temperature.

The immunostaining was performed on a Leica Biosystems BOND^® RX instrument.

Rabbit specific IHC polymer detection kit HRP/DAB (ab209101) was used as the secondary detection system.

Counterstained with Hematoxylin.

Heat mediated antigen retrieval was performed with Citrate buffer (pH 6.0, Epitope Retrieval Solution 1) for 20 mins.

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-gamma Synuclein/SNCG antibody [EP1539Y] (AB52633)

Immunofluorescent analysis of 4% Paraformaldehyde fixed 0.1% Triton X-100 permeabilized wild-type HeLa (human cervical adenocarcinoma epithelial cell) and gamma Synuclein/SNCG knockout HeLa cells (ab265623) labelling gamma Synuclein/SNCG with the target antibody at 1/50 (2.06 μg/ml) followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) preadsorbed antibody as secondary at 1/1000 (2 μg/ml) dilution. Confocal image showing partial nuclear and cytoplasmic staining in wild-type HeLa cells (shown in green), with no staining observed in gamma Synuclein/SNCG knockout HeLa cells. ab7291 Anti-alpha Tubulin mouse monoclonal antibody at 1/1000 (1 μg/ml) followed by ab150120 Goat Anti-Mouse IgG H&L (Alexa Fluor^® 594) at 1/1000 (2 μg/ml) dilution was used as a counterstain (shown in magenta). Nuclear DNA was labelled with DAPI (shown in blue). Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-gamma Synuclein/SNCG antibody [EP1539Y] (AB52633)

ab52633 staining human brain in IHC-P.

Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

• WB

Lab

Western blot - Anti-gamma Synuclein/SNCG antibody [EP1539Y] (AB52633)

Lanes 1-3 : Merged signal (red and green). Green - ab52633 observed at 16 kDa. Red - loading control ab8245 observed at 36 kDa.

ab52633 Anti-gamma Synuclein/SNCG antibody [EP1539Y] was shown to specifically react with gamma Synuclein/SNCG antibody in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab265623 (knockout cell lysate ab257694) was used. Wild-type and gamma Synuclein/SNCG knockout samples were subjected to SDS-PAGE. ab52633 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C at 1 in 1000 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye^® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preadsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.

All lanes:

Western blot - Anti-gamma Synuclein/SNCG antibody [EP1539Y] (ab52633) at 1/1000 dilution

Lane 1:

Wild-type HeLa cell lysate at 20 µg

Lane 2:

SNCG knockout HeLa cell lysate at 20 µg

Lane 2:

Western blot - Human SNCG (gamma Synuclein) knockout HeLa cell line (<a href='/ko/products/cell-lines/human-sncg-gamma-synuclein-knockout-hela-cell-line-ab265623'>ab265623</a>)

Lane 3:

SK-BR-3 cell lysate at 20 µg

Secondary

All lanes:

Western blot - Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (<a href='/ko/products/secondary-antibodies/goat-rabbit-igg-h-l-irdye-800cw-preadsorbed-ab216773'>ab216773</a>) at 1/10000 dilution

Predicted band size: 13 kDa

Observed band size: 16 kDa

false

• WB

Unknown

Western blot - Anti-gamma Synuclein/SNCG antibody [EP1539Y] (AB52633)

All lanes:

Western blot - Anti-gamma Synuclein/SNCG antibody [EP1539Y] (ab52633) at 1/2000 dilution

All lanes:

Human fetal brain lysate

Predicted band size: 13 kDa

false