Anti-Alpha-synuclein (phospho S129) antibody [EP1536Y]

12 product images

• 

  This image is taken from Machado de Oliveira. R et al. PLoS Biol. 2017 Mar; 15(3): e2000374. S7a doi: 10.1371/journal.pbio.2000374 Reproduced under the Creative Commons license http://creativecommons.org/licenses/by/4.0/.

  Immunohistochemistry - Free Floating - Anti-Alpha-synuclein (phospho S129) antibody [EP1536Y] (ab51253)

  Phospho alpha-synuclein immunohistochemistry staining of mouse brain using rabbit anti-Phospho alpha-synuclein antibody
  
  Expression of WT aSyn in the SN is toxic over time. Mouse brain sections immunostained for TH (red panels) and aSyn (using ab51253) (green panels) 1 2 and 3 weeks after injection with vectors encoding for EGFP or WT aSyn. Scale bar for isolated channels 200 μm and for merged channels 100 μm.

• 

  This image is taken from Migdalska-Richards A. et al. Ann Neurol. 2016 Nov; 80(5): 766–775. Fig 5 doi: 10.1002/ana.24790

  Western blot - Anti-Alpha-synuclein (phospho S129) antibody [EP1536Y] (ab51253)

  α‐Synuclein (α‐syn) and S129‐phosphorylated α‐synuclein protein levels in SNCA/SNCA mouse brains after 12 days of treatment with 4mM ambroxol (Amb). (A) Western blotting for α‐synuclein (using Anti-Alpha-synuclein antibody [4D6] - BSA and Azide free ab1903) and serine 129 (S129)‐phosphorylated α‐synuclein protein (using ab51253) in the brainstem (example blots shown).

  All lanes: Western blot - Anti-Alpha-synuclein (phospho S129) antibody [EP1536Y] (ab51253)

  Predicted band size: 14 kDa

• 

  Western blot - Anti-Alpha-synuclein (phospho S129) antibody [EP1536Y] (ab51253)

  Blocking and Diluting buffer and concentration - 5% NFDM/TBST
  

  Exposure time: 3 seconds

  All lanes: Western blot - Anti-Alpha-synuclein (phospho S129) antibody [EP1536Y] (ab51253) at 1/1000 dilution

  Lane 1: In vitro kinase assay of Alpha Synuclein phosphorylation using His tagged human full length recombinant alpha-synuclein protein in the presence of PLK2 (Polo-like kinase 2) but absence of AT

  Lane 2: In vitro kinase assay of Alpha Synuclein phosphorylation using His tagged human full length recombinant alpha-synuclein protein in the presence of ATP but absence of PLK2 (Polo-like kinase 2

  Lane 3: In vitro kinase assay of Alpha Synuclein phosphorylation using His tagged human full length recombinant alpha-synuclein protein in the presence of PLK2 (Polo-like kinase 2) and ATP

  Secondary

  All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/20000 dilution

  Predicted band size: 14 kDa

  Observed band size: 17 kDa

• 

  Western blot - Anti-Alpha-synuclein (phospho S129) antibody [EP1536Y] (ab51253)

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

  Exposure time: 20 seconds

  All lanes: Western blot - Anti-Alpha-synuclein (phospho S129) antibody [EP1536Y] (ab51253) at 1/1000 dilution

  Lane 1: Rat brain lysates, the membrane was incubated with alkaline phosphatase. at 15 µg

  Lane 2: Rat brain lysates at 15 µg

  Secondary

  All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/2000 dilution

  Predicted band size: 14 kDa

  Observed band size: 18 kDa

• 

  Western blot - Anti-Alpha-synuclein (phospho S129) antibody [EP1536Y] (ab51253)

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

  Exposure time: 40 seconds

  All lanes: Western blot - Anti-Alpha-synuclein (phospho S129) antibody [EP1536Y] (ab51253) at 1/1000 dilution

  Lane 1: Mouse brain lysates, the membrane was incubated with alkaline phosphatase at 15 µg

  Lane 2: Mouse brain lysates at 15 µg

  Secondary

  All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/2000 dilution

  Predicted band size: 14 kDa

  Observed band size: 18 kDa

• 

  This image is courtesy of Professor Michael Goedert’s team (Sophie Morgan and Masami Masuda-Suzukake).

  Western blot - Anti-Alpha-synuclein (phospho S129) antibody [EP1536Y] (ab51253)

  All lanes: Western blot - Anti-Alpha-synuclein (phospho S129) antibody [EP1536Y] (ab51253) at 1/1000 dilution

  Lane 1: Sarkosyl-insoluble brain extract from mice transgenic for PrPA53T alpha-synuclein (line M83)

  Lane 2: Recombinant alpha-synuclein phosphorylated at S129, 3ng

  Predicted band size: 14 kDa

• 

  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Alpha-synuclein (phospho S129) antibody [EP1536Y] (ab51253)

  IHC image of alpha Synuclein (phospho S129) staining Human Parkinson Substantia Nigra tissue section*, previously antigen was retrieved by heat mediated with citrate buffer pH 6, fixed in formalin and embedded in paraffin. This section was incubated with ab51253 at 10 μg/mL for 15 mins at room temperature and detected using an HRP conjugated compact polymer system, performed on a Leica Bond™ system using the standard protocol F. DAB was used as the chromogen. Counterstained with haematoxylin and mounted with DPX. The inset negative control image is taken from an identical assay performed on Human normal Substantia Nigra.
  

  For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.

  *Tissue obtained from the Human Research Tissue Bank, supported by the NIHR Cambridge Biomedical Research Centre

• 

  Dot Blot - Anti-Alpha-synuclein (phospho S129) antibody [EP1536Y] (ab51253)

  Dot blot analysis of human alpha Synuclein (pS129) peptide (Lane 1), huamn alpha Synuclein (unmodified) peptide (Lane 2) labelling alpha Synuclein (pS129) with ab51253 at a dilution of 1/1000. Peroxidase conjugated goat anti-rabbit IgG (H+L) was used as the secondary antibody at a dilution of 1/2500.
  

  Blocking and dilution buffer: 5% NFDM/TBST.

  Exposure time: 3 minutes.

• 

  This image is courtesy of a customer review by Sophie Morgan

  Immunohistochemistry - Free Floating - Anti-Alpha-synuclein (phospho S129) antibody [EP1536Y] (ab51253)

  Phospho alpha-synuclein immunohistochemistry staining of mouse brain using rabbit anti-Phospho alpha-synuclein antibody
  
  IHC image of alpha Synuclein (phospho S129) staining in free floating tgM83^+/- or tgM83^+/+ mouse brain tissue. The section was incubated with ab51253 at 1/5000 dilution for 15 hours at 4^oC and detected using a Biotinylated conjugated Anti-Rabbit monocloal antibody at 1/200 dilution.

• 

  ELISA - Anti-Alpha-synuclein (phospho S129) antibody [EP1536Y] (ab51253)

  Direct ELISA antibody dose-response curve using ab51253.
  

  Antibody concentration of 0-5000 ng/mL.

  Antigen (Alpha-synuclein (pS129) phospho peptide and Alpha-synuclein non-phospho peptide) concentration of 1000 ng/mL.

  An alkaline phosphatase conjugated goat anti-rabbit IgG (1/1000) was used as the secondary antibody.

• 

  Western blot - Anti-Alpha-synuclein (phospho S129) antibody [EP1536Y] (ab51253)

  Blocking and diluting buffer and concentration: 5% NFDM/TBST
  
  This blot was developed using a higher sensitivity ECL substrate.
  
  Band around 100kda corresponds to αS oligomer (PMID: 27637918, PMID: 12597857)
  
  Lysates used here were prepared using RIPA method. We recommend 1% SDS hot lysate method to reduce the detection of oligomers. Please refer to Support & downloads -> Downloads -> Protocol booklet.

  All lanes: Western blot - Anti-Alpha-synuclein (phospho S129) antibody [EP1536Y] (ab51253) at 1/1000 dilution

  Lane 1: Mouse brain with Alzheimer’s disease tissue lysate at 20 µg

  Lane 2: Mouse brain with Alzheimer’s disease tissue lysate, membrane was incubated with alkaline phosphatase

  Secondary

  All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/20000 dilution

  Predicted band size: 14 kDa

  Observed band size: 18 kDa, 100 kDa

• 

  Immunohistochemistry paraffin embedded sections - Anti-Alpha-synuclein (phospho S129) antibody [EP1536Y] (ab51253)

  Immunohistochemistry image of Alpha Synuclein Phospho S129 staining in a section of formalin-fixed paraffin-embedded normal human cerebral cortex* performed on a Leica Biosystems BOND^® RX instrument using the standard protocol. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20mins. The section was then incubated with ab51253, 5 µg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX. The inset secondary-only control image is taken from an identical assay without primary antibody.

  For other Immunohistochemistry staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.

  *Tissue obtained from the Human Research Tissue Bank, supported by the NIHR Cambridge Biomedical Research Centre.