Recombinant Anti-Tau (phospho S396) antibody [EPR2731] (ab109390)

Blocking buffer: 5% NFDM/TBST

Dilution buffer: 5% NFDM/TBST

IHC image of Tau (phospho S396) staining in a section of frozen normal human Alzheimer brain performed on a Leica BOND^TM system using the standard protocol. The section was fixed in 10% paraformaldehyde (10 min) prior to staining. The section was incubated with ab109390, 1/1000 dilution, 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 IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.

Immunohistochemistry (Frozen sections) analysis of rat cerebrum tissue sections labeling Tau with Purified ab109390 at 1/100 (1.0 µg/ml).Heat mediated antigen retrieval using sodium citrate buffer (10mM citrate pH 6.0 + 0.05% Tween-20). Goat anti rabbit IgG (Alexa Fluor^® 488, ab150077) was used as the secondary antibody. Negative control: PBS instead of the primary antibody. DAPI was used as a counterstain.

Immunohistochemical staining of paraffin embedded human glioblastoma with purified ab109390 at a dilution of 1/4000. A pre-diluted HRP polymer for rabbit/mouse IgG was used as the secondary antibody and the sample was counter-stained wirh hematoxylin. Antigen retrieval was perfomed using Tris-EDTA buffer, pH 9.0. PBS was used instead of the primary antibody as the negative control, and is shown in the inset.

ab109390 at 1/20 immunoprecipitating Tau (phospho S396) in Human brain lysate.

Lane 1 (input): Human brain lysate (10µg)

Lane 2 (+): ab109390 + Human brain lysate.

Lane 3 (-): Rabbit monoclonal IgG (ab172730) instead of ab109390 in Human brain lysate.

For western blotting, ab109390  at 1/1000 dilution followed by VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/5000 dilution.

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

Immunohistochemistry (Frozen sections) analysis of mouse cerebrum tissue sections labeling Tau with Purified ab109390 at 1/100 (1.0 µg/ml).Heat mediated antigen retrieval using sodium citrate buffer (10mM citrate pH 6.0 + 0.05% Tween-20). Goat anti rabbit IgG (Alexa Fluor^® 488, ab150077) was used as the secondary antibody. Negative control: PBS instead of the primary antibody. DAPI was used as a counterstain.

IHC image of Tau (phospho S396) staining in human Alzheimer hippocampus formalin fixed paraffin embedded tissue section. The section was pre-treated using heat mediated antigen retrieval with citrate buffer. The section was then incubated with unpurified ab109390 at 1/1000 dilution for 2 hoursat 21ºC. A biotin conjugated goat-anti-rabbit antibody was used as a secondary at 1/250. The section shows clear neurofibrillary tangles in a subset of neurons.

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Diluting and blocking buffer: 5% NFDM/TBST

Blocking buffer: 5% NFDM/TBST

Dilution buffer: 5% NFDM/TBST

Dot blot analysis of Tau (phospho S396) phospho peptide (Lane 1) and Tau non-phospho peptide (Lane 2) labeling Tau (phospho S396) with ab109390 at a dilution of 1/1000. ab97051 (Peroxidase conjugated goat anti-rabbit IgG) (H+L) at 1/100 000 was used as the secondary antibody.

Blocking and diluting buffer: 5% NFDM/TBST.

Exposure time: 3 minutes.