Anti-Alpha-synuclein (N-term) antibody [EPR29522-6]

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Alpha-synuclein (N-term) antibody [EPR29522-6] (AB324977)

Immunohistochemical analysis of paraffin-embedded Human hypothalamus tissue labeling Alpha-synuclein (N-term) with ab324977 at 1/5000 (0.106 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on human hypothalamus.
The primary antibody was incubated 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 was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Alpha-synuclein (N-term) antibody [EPR29522-6] (AB324977)

Immunohistochemical analysis of paraffin-embedded Human cerebrum tissue labeling Alpha-synuclein (N-term) with ab324977 at 1/5000 (0.106 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on human cerebrum.
The primary antibody was incubated 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 was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Alpha-synuclein (N-term) antibody [EPR29522-6] (AB324977)

Immunohistochemical analysis of paraffin-embedded Human Parkinson's disease brain tissue labeling Alpha-synuclein (N-term) with ab324977 at 1/5000 (0.106 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on human Parkinson's disease brain.
The primary antibody was incubated 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 was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Alpha-synuclein (N-term) antibody [EPR29522-6] (AB324977)

Immunohistochemical analysis of paraffin-embedded Mouse cerebrum tissue labeling Alpha-synuclein (N-term) with ab324977 at 1/5000 (0.106 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on mouse cerebrum.
The primary antibody was incubated 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 was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins

• WB

Supplier Data

Western blot - Anti-Alpha-synuclein (N-term) antibody [EPR29522-6] (AB324977)

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

Low expression : liver, kidney(PMID : 8194594)

The expression profile/ molecular weight observed is consistent with what has been described in the literature (PMID : 11739566; PMID : 22112368; PMID : 14997013).

Lanes 1-4 are incubated with are incubated with Goat Anti-Rabbit IgG (HRP) with minimal cross-reactivity with human IgG at 1/2000 and lanes 5-8 are incubated with Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (ab97051) at 1/20000.

The identity of the higher MW band at approximately 50 kDa (in lane 2, 5 and lanes 7- 8) is unknown.

In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) staining at 1/200000 dilution.

All lanes:

Western blot - Anti-Alpha-synuclein (N-term) antibody [EPR29522-6] (ab324977) at 1/1000 dilution

Lane 1:

Human cerebellum tissue lysate at 20 µg

Lane 2:

Human liver tissue lysate at 20 µg

Lane 3:

Human hypothalamus tissue lysate at 20 µg

Lane 4:

Human thalamus tissue lysate at 20 µg

Lane 5:

Mouse brain tissue lysate at 20 µg

Lane 6:

Mouse liver tissue lysate at 20 µg

Lane 7:

Mouse hippocampus tissue lysate at 20 µg

Lane 8:

Mouse kidney tissue lysate at 20 µg

Secondary

Lanes 1 - 4:

Goat Anti-Rabbit IgG (HRP) with minimal cross-reactivity with human IgG at 1/2000 dilution

Lanes 5 - 8:

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

Observed band size: 18 kDa,36 kDa

false

Exposure time: 103s

• WB

Supplier Data

Western blot - Anti-Alpha-synuclein (N-term) antibody [EPR29522-6] (AB324977)

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

Negative control : A549, HepG2.

The expression profile/ molecular weight observed is consistent with what has been described in the literature (PMID : 11739566; PMID : 22112368; PMID : 14997013).

The identity of the higher MW band at approximately 50 kDa (in lane 2) is unknown.

In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) staining at 1/200000 dilution.

All lanes:

Western blot - Anti-Alpha-synuclein (N-term) antibody [EPR29522-6] (ab324977) at 1/1000 dilution

Lane 1:

G-401 (human rhabdoid tumor kidney epithelial cell) whole cell lysate at 20 µg

Lane 2:

A549 (human lung carcinoma epithelial cell) whole cell lysate at 20 µg

Lane 3:

HepG2 (human hepatocellular carcinoma epithelial cell) whole cell lysate at 20 µg

Lane 4:

SK-MEL-28 (human malignant melanoma cell ) whole cell lysate at 20 µg

Secondary

All lanes:

Western blot at 1/20000 dilution

Observed band size: 18 kDa,36 kDa

false

Exposure time: 180s

• WB

Supplier Data

Western blot - Anti-Alpha-synuclein (N-term) antibody [EPR29522-6] (AB324977)

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

In Western blot, ab324977 was shown to bind specifically to Alpha-synuclein. Target of interest was observed at 18 kDa in wild-type HAP1 cell lysates (lane 1) with no signal observed at this size in Alpha-synuclein knockout cell line (lane 2).

In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) staining at 1/200000 dilution.

All lanes:

Western blot - Anti-Alpha-synuclein (N-term) antibody [EPR29522-6] (ab324977) at 1/1000 dilution

Lane 1:

Wild-type HAP1 (human chronic myelogenous leukemia near-haploid cell ) whole cell lysate at 20 µg

Lane 2:

Alpha-synuclein knockout HAP1 whole cell lysate at 20 µg

Secondary

All lanes:

Western blot at 1/20000 dilution

Observed band size: 18 kDa,36 kDa

false

Exposure time: 180s

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Alpha-synuclein (N-term) antibody [EPR29522-6] (AB324977)

Immunohistochemical analysis of paraffin-embedded Human liver tissue labeling Alpha-synuclein (N-term) with ab324977 at 1/5000 (0.106 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Low expression tissue : No staining on human liver (PMID : 8194594).
The primary antibody was incubated 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 was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Alpha-synuclein (N-term) antibody [EPR29522-6] (AB324977)

Immunohistochemical analysis of paraffin-embedded Mouse liver tissue labeling Alpha-synuclein (N-term) with ab324977 at 1/5000 (0.106 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Low expression tissue : No staining on mouse liver.
The primary antibody was incubated 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 was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins

• ELISA

Supplier Data

ELISA - Anti-Alpha-synuclein (N-term) antibody [EPR29522-6] (AB324977)

Indirect ELISA analysis of ab324977 at 1000-0 ng/ml. The Secondary antibody used was Alkaline Phosphatase-conjugated AffiniPure Goat Anti-Rabbit IgG (H+L) at 1 : 2500 dilution dilution.

Antigen : Human Alpha-synuclein protein;Human Alpha-synuclein (N-term) peptide;Human Beta-synuclein protein.

Antigen concentration : 1000 ng/ml

• WB

Supplier Data

Western blot - Anti-Alpha-synuclein (N-term) antibody [EPR29522-6] (AB324977)

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

This antibody does not cross-react with human Beta-synuclein.

In Western blot, Anti-6X His tag® antibody [AD1.1.10] (ab15149) staining at 1/1000 dilution.

All lanes:

Western blot - Anti-Alpha-synuclein (N-term) antibody [EPR29522-6] (ab324977) at 1/1000 dilution

Lane 1:

His-tagged human Alpha-synuclein recombinant protein at 10 ng

Lane 2:

His-tagged human Beta-synuclein recombinant protein at 10 ng

Secondary

All lanes:

Western blot at 1/20000 dilution

Observed band size: 18 kDa

false

Exposure time: 180s