Anti-Alpha-synuclein antibody [MJFR1] - Chicken IgY (Chimeric) - BSA and Azide free

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Alpha-synuclein antibody [MJFR1] - Chicken IgY (Chimeric) - BSA and Azide free (AB320840)

This data was developed using ab320839, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin-embedded (A) Parental HAP1 (human chronic myelogenous leukemia near-haploid cell) cell pellet (B) SNCA knockout HAP1 (HO010074) cell pellet labeling Alpha-synuclein with ab320839 at 1/1000 (1.055 ug/ml) dilution, 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 ab320839 for 10 mins at room temperature, followed by anti-Chicken IgY (ab97136) 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] - Chicken IgY (Chimeric) - BSA and Azide free (AB320840)

This data was developed using ab320839, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin-embedded Human liver tissue labeling Alpha-synuclein with ab320839 at 1/5000 (0.211 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Negative control : no staining on human liver.
The section was incubated with ab320839 for 10 mins at room temperature, followed by anti-Chicken IgY antibody (ab97136) 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] - Chicken IgY (Chimeric) - BSA and Azide free (AB320840)

This data was developed using ab320839, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin-embedded Human cerebrum tissue labeling Alpha-synuclein with ab320839 at 1/5000 (0.211 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on human cerebrum.
The section was incubated with ab320839 for 10 mins at room temperature, followed by anti-Chicken IgY antibody (ab97136) 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

Supplier Data

Western blot - Anti-Alpha-synuclein antibody [MJFR1] - Chicken IgY (Chimeric) - BSA and Azide free (AB320840)

This data was developed using ab320839, the same antibody clone in a different buffer formulation.

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

Negative control : liver and kidney ( (PMID : 14997013).

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

All lanes:

Western blot - Anti-Alpha-synuclein antibody [MJFR1] - Chicken IgY (Chimeric) (<a href='/ko/products/primary-antibodies/alpha-synuclein-antibody-mjfr1-chicken-igy-chimeric-ab320839'>ab320839</a>) 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 kidney tissue lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Chicken IgY H&L (HRP) preadsorbed (<a href='/ko/products/secondary-antibodies/goat-chicken-igy-h-l-hrp-preadsorbed-ab7118'>ab7118</a>) at 1/2000 dilution

Observed band size: 18 kDa,36 kDa

false

Exposure time: 180s

• WB

Supplier Data

Western blot - Anti-Alpha-synuclein antibody [MJFR1] - Chicken IgY (Chimeric) - BSA and Azide free (AB320840)

This data was developed using ab320839, the same antibody clone in a different buffer formulation.

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

In Western blot, ab320839 was shown to bind specifically to SNCA. Target of interest was observed at 18kDa in wild-type HAP1 cell lysates (lane 1) with no signal observed at this size in SNCA knockout cell line (lane 2).

The identity of the bands above target band are unknown.

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

All lanes:

Western blot - Anti-Alpha-synuclein antibody [MJFR1] - Chicken IgY (Chimeric) (<a href='/ko/products/primary-antibodies/alpha-synuclein-antibody-mjfr1-chicken-igy-chimeric-ab320839'>ab320839</a>) at 1/1000 dilution

Lane 1:

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

Lane 2:

SNCA knockout HAP1 whole cell lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Chicken IgY H&L (HRP) preadsorbed (<a href='/ko/products/secondary-antibodies/goat-chicken-igy-h-l-hrp-preadsorbed-ab7118'>ab7118</a>) at 1/2000 dilution

Observed band size: 18 kDa,36 kDa

true

Exposure time: 180s