Anti-Alpha-synuclein antibody [MJFR1] - Mouse IgG2a (Chimeric)

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Alpha-synuclein antibody [MJFR1] - Mouse IgG2a (Chimeric) (AB309157)

Immunohistochemical analysis of paraffin-embedded human cerebrum tissue labeling Alpha-synuclein with ab309157 at 1/50000 (0.017 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Cytoplasmic staining on human cerebrum. The section was incubated with ab309157 for 10 mins at room temperature, followed by anti-mouse IgG2a antibody for 8 mins during the LeicaDS9800 kit staining procedure. 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 Citrate buffer (pH 6.0, Epitope Retrieval Solution 1) for 20 mins.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Alpha-synuclein antibody [MJFR1] - Mouse IgG2a (Chimeric) (AB309157)

Immunohistochemical analysis of paraffin-embedded cells labeling Alpha-synuclein with ab309157 at 1/500 (1.7 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Cytoplasmic staining on (A) Parental HAP1 cell pellet, no staining on (B) SNCA knockout HAP1 (HO010074) cell pellet. The section was incubated with ab309157 for 10 mins at room temperature, followed by anti-mouse IgG2a antibody for 8 mins during the LeicaDS9800 kit staining procedure. 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 Citrate buffer (pH 6.0, Epitope Retrieval Solution 1) for 20 mins.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Alpha-synuclein antibody [MJFR1] - Mouse IgG2a (Chimeric) (AB309157)

Immunohistochemical analysis of paraffin-embedded human kidney tissue labeling Alpha-synuclein with ab309157 at 1/50000 (0.017 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Negative control : No staining on human kidney. The section was incubated with ab309157 for 10 mins at room temperature, followed by anti-mouse IgG2a antibody for 8 mins during the LeicaDS9800 kit staining procedure. 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 Citrate buffer (pH 6.0, Epitope Retrieval Solution

1. for 20 mins.

• WB

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Western blot - Anti-Alpha-synuclein antibody [MJFR1] - Mouse IgG2a (Chimeric) (AB309157)

Blocking and diluting buffer and concentration : 5% NFDM/TBST. Negative control : Kidney and heart. (PMID : 14997013). In Western blot, anti-GAPDH antibody (ab181602) loading control staining at 1/200000 dilution. Exposure time : 37 seconds.

All lanes:

Western blot - Anti-Alpha-synuclein antibody [MJFR1] - Mouse IgG2a (Chimeric) (ab309157) at 1/1000 dilution

Lane 1:

Human cerebellum tissue lysate at 10 µg

Lane 2:

Human heart tissue lysate at 10 µg

Lane 3:

Human kidney tissue lysate at 10 µg

Secondary

All lanes:

Western blot - Goat Anti-Mouse IgG Fc (HRP) preadsorbed (<a href='/en-us/products/secondary-antibodies/goat-mouse-igg-fc-hrp-preadsorbed-ab98717'>ab98717</a>) at 1/20000 dilution

Observed band size: 18 kDa

false

Exposure time: 37s

• WB

Supplier Data

Western blot - Anti-Alpha-synuclein antibody [MJFR1] - Mouse IgG2a (Chimeric) (AB309157)

Blocking and diluting buffer and concentration : 5% NFDM/TBST. In Western blot, ab309157 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 (ab181602) loading control staining at 1/200000 dilution. Exposure time : 180 seconds.

All lanes:

Western blot - Anti-Alpha-synuclein antibody [MJFR1] - Mouse IgG2a (Chimeric) (ab309157) at 1/1000 dilution

Lane 1:

Parental HAP1 whole cell lysate at 20 µg

Lane 2:

Alpha-synuclein knockout HAP1 whole cell lysate at 20 µg

Secondary

All lanes:

Peroxidase-Conjugated Goat anti-Mouse IgG (H+L) at 1/5000 dilution

Observed band size: 18 kDa

false

Exposure time: 180s