Anti-beta Synuclein antibody [EP1537Y] - BSA and Azide free

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-beta Synuclein antibody [EP1537Y] - BSA and Azide free (AB221908)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human cerebrum tissue sections labeling beta Synuclein with purified ab76111 at 1/50 dilution (2.4 µg/mL). Perform heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0). ImmunoHistoProbe one step HRP Polymer (ready to use) was used as the secondary antibody. Negative control : PBS instead of the primary antibody. Hematoxylin was used as a counterstain.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab76111).

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-beta Synuclein antibody [EP1537Y] - BSA and Azide free (AB221908)

Immunohistochemical analysis of paraffin-embedded Human cerebrum tissue labeling beta Synuclein with ab221908 at 1/5000 dilution. followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin. Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20 mins.

• IP

Unknown

Immunoprecipitation - Anti-beta Synuclein antibody [EP1537Y] - BSA and Azide free (AB221908)

Purified ab76111 at 1/20 dilution (0.6 µg) immunoprecipitating beta Synuclein in Human brain lysate.
Lane 1 (input) : Human brain lysate 10 µg
Lane 2 (+) : ab76111 + Human brain lysate.
Lane 3 (-) : Rabbit monoclonal IgG (ab172730) instead of ab76111 in Human brain lysate.
VeriBlot for IP Detection Reagent (HRP) (ab131366) (1/1000 dilution) was used for Western blotting.
Blocking Buffer and concentration : 5% NFDM/TBST.
Diluting buffer and concentration : 5% NFDM/TBST.
Observed band size : 18 kDa
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab76111).

All lanes:

Immunoprecipitation - Anti-beta Synuclein antibody [EP1537Y] (<a href='/en-us/products/primary-antibodies/beta-synuclein-antibody-ep1537y-ab76111'>ab76111</a>)

Predicted band size: 14 kDa

false

• Flow Cyt (Intra)

Unknown

Flow Cytometry (Intracellular) - Anti-beta Synuclein antibody [EP1537Y] - BSA and Azide free (AB221908)

Intracellular Flow Cytometry analysis of PC-12 (Rat adrenal gland pheochromocytoma) cells labeling beta Synuclein with purified ab76111 at 1/100 dilution (10 μg/mL) (Red). Cells were fixed with 4% Paraformaldehyde and permeabilised with 90% Methanol. A Goat anti rabbit IgG (Alexa Fluor^® 488, ab150077) secondary antibody was used at 1/2000. Isotype control - Rabbit monoclonal IgG (Black). Unlabeled control - Cell without incubation with primary antibody and secondary antibody (Blue).

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab76111).

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-beta Synuclein antibody [EP1537Y] - BSA and Azide free (AB221908)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of mouse cerebrum tissue sections labeling beta Synuclein with purified ab76111 at 1/50 dilution (2.4 µg/mL). Perform heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0). ImmunoHistoProbe one step HRP Polymer (ready to use) was used as the secondary antibody. Negative control : PBS instead of the primary antibody. Hematoxylin was used as a counterstain.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab76111).

• Flow Cyt (Intra)

Supplier Data

Flow Cytometry (Intracellular) - Anti-beta Synuclein antibody [EP1537Y] - BSA and Azide free (AB221908)

PC-12 cells were fixed with 100µl 4% PFA for 10 min at RT, then permeabilized with 100µl 90% methanol for 30 min (-20° C) followed by blocking with 10% goat serum for 1h at room temperature. Incubate ab221908 for 30 min at room temperature (RT) and then incubate Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150081) for 30 min at room temperature.

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-beta Synuclein antibody [EP1537Y] - BSA and Azide free (AB221908)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized PC-12 cells labelling beta Synuclein with ab221908 at 1/100 dilution, followed by AlexaFluor® 488-conjugated Goat anti-Rabbit IgG H&L preadsorbed secondary antibody (ab150081) at 1/1000 dilution at RT for 45 min (Green). ab195889 Anti-alpha Tubulin, used as a counterstain at 1/200 dilution, was co-incubated with ab221908 overnight at 4° C. Nucleus were visualized using DAPI.

• ICC/IF

Unknown

Immunocytochemistry/ Immunofluorescence - Anti-beta Synuclein antibody [EP1537Y] - BSA and Azide free (AB221908)

Immunocytochemistry/ Immunofluorescence analysis of PC-12 (Rat adrenal gland pheochromocytoma) cells labeling beta Synuclein with purified ab76111 at 1/50 dilution (2.4 μg/mL). Cells were fixed in 4% Paraformaldehyde and permeabilized with 0.1% TritonX-100. Cells were counterstained with ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor^® 594) 1/200 (2.5 μg/mL). Goat anti rabbit IgG (Alexa Fluor^® 488, ab150077) was used as the secondary antibody at 1/1000 (2 μg/mL) dilution. DAPI (blue) was used as nuclear counterstain. PBS instead of the primary antibody was used as the secondary antibody only control.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab76111).

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-beta Synuclein antibody [EP1537Y] - BSA and Azide free (AB221908)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of rat cerebrum tissue sections labeling beta Synuclein with purified ab76111 at 1/50 dilution (2.4 µg/mL). Perform heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0). ImmunoHistoProbe one step HRP Polymer (ready to use) was used as the secondary antibody. Negative control : PBS instead of the primary antibody. Hematoxylin was used as a counterstain.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab76111)

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-beta Synuclein antibody [EP1537Y] - BSA and Azide free (AB221908)

Immunohistochemical analysis of paraffin-embedded Rat cerebrum tissue labeling beta Synuclein with ab221908 at 1/5000 dilution. followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin. Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20 mins.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-beta Synuclein antibody [EP1537Y] - BSA and Azide free (AB221908)

Immunohistochemical analysis of paraffin-embedded Mouse cerebrum tissue labeling beta Synuclein with ab221908 at 1/5000 dilution. followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin. Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20 mins.

• IP

Supplier Data

Immunoprecipitation - Anti-beta Synuclein antibody [EP1537Y] - BSA and Azide free (AB221908)

beta Synuclein was immunoprecipitated from 0.35 mg of mouse brain tissue lysate with ab221908 at 1/30 dilution. Western blot was performed from the immunoprecipitated using ab221908 at 1/1000 dilution.

Lane 1:

Mouse brain tissue lysate (Input) at 10 µg

Lane 2:

Mouse brain tissue lysate (+)

Lane 3:

Rabbit monoclonal IgG (<a href='/en-us/products/primary-antibodies/rabbit-igg-monoclonal-epr25a-isotype-control-ab172730'>ab172730</a>) instead of ab221908 in HepG2 whole cell lysate (-)

Secondary

All lanes:

Immunoprecipitation - VeriBlot for IP Detection Reagent (HRP) (<a href='/en-us/products/reagents/veriblot-for-ip-detection-reagent-hrp-ab131366'>ab131366</a>) at 1/5000 dilution

false

• WB

Unknown

Western blot - Anti-beta Synuclein antibody [EP1537Y] - BSA and Azide free (AB221908)

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab76111).

All lanes:

Western blot - Anti-beta Synuclein antibody [EP1537Y] (<a href='/en-us/products/primary-antibodies/beta-synuclein-antibody-ep1537y-ab76111'>ab76111</a>) at 1/10000 dilution

Lane 1:

Human brain lysate at 20 µg

Lane 2:

Mouse brain lysate at 20 µg

Lane 3:

Rat brain lysate at 20 µg

Secondary

All lanes:

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

Predicted band size: 14 kDa

Observed band size: 18 kDa

false

• WB

Unknown

Western blot - Anti-beta Synuclein antibody [EP1537Y] - BSA and Azide free (AB221908)

A : Anti-beta Synuclein antibody [EP1537Y] (ab76111)

B : Anti-Alpha-synuclein antibody [MJFR1] (ab138501)

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab76111).

All lanes:

Western blot - Anti-beta Synuclein antibody [EP1537Y] (<a href='/en-us/products/primary-antibodies/beta-synuclein-antibody-ep1537y-ab76111'>ab76111</a>) at 1/10000 dilution

Lane 1:

His-Tagged human beta Synuclein (aa 1 to 134) recombinant protein at 0.015 µg

Lane 2:

His-Tagged alpha synuclein (aa 1 to 140) recombinant protein at 0.015 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

Supplier Data

Western blot - Anti-beta Synuclein antibody [EP1537Y] - BSA and Azide free (AB221908)

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

Lane 1:

His-Tagged human beta Synuclein (aa 1 to 134) recombinant protein at 15 µg

Lane 2:

His-Tagged human alpha synuclein (aa 1 to 140) recombinant protein at 15 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

false