Anti-Tau (phospho T231) antibody [RP23040023]

5 product images

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  Western blot - Anti-Tau (phospho T231) antibody [RP23040023] (ab307987)

  Western blot analysis of Tau (phospho T231)was performed by rat brain lysate (lane 1) using 4-12% Bis-Tris gel, electrophoresis system with a pre-stained protein standard. Proteins were transferred to a nitrocellulose membrane and blocked with 5% skim milk for 1 hour at room temperature. Tau (pT231) was detected at ~55 kDa using ab307987 at a 1/1000 dilution in 2.5% skim milk at 4°C overnight on a rocking platform. To confirm specificity, competition was performed with the phosphopeptide (10 µg/mL) (lane 2). Detection was performed using an HRP-conjugated Goat anti-Rabbit secondary antibody at a 1/5000 dilution and chemiluminescent detection was performed (ECL).

  All lanes: Western blot - Anti-Tau (phospho T231) antibody [RP23040023] (ab307987) at 1/1000 dilution

  Lane 1: rat brain lysate at 20 µg

  Lane 2: rat brain lysate with phosphopeptide at 20 µg

  Secondary

  All lanes: HRP-conjugated Goat anti-Rabbit secondary antibody at 1/5000 dilution

  Developed using the ECL technique.

  Observed band size: 55 kDa

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  Immunocytochemistry/ Immunofluorescence - Anti-Tau (phospho T231) antibody [RP23040023] (ab307987)

  For immunofluorescence analysis, 70% confluent log phase HeLa cells were fixed with 4% paraformaldehyde for 15 minutes, permeabilized with 0. 25% Triton X-100 for 10 minutes, and blocked with 5% BSA for 1 hour at room temperature then stained for Tau (phospho T231) using ab307987 at 1/500 dilution and incubated for 3 hours at room temperature, then labeled with Alexa Fluor® 488 Goat anti-Rabbit IgG secondary antibody at a dilution of 1/400 for 30 minutes at room temperature (Panel a: green).
  
  Nuclei were stained with DAPI (Panel b: blue).
  
  F-actin was stained with Alexa Fluor® 594 phalloidin (Panel c: red)
  
  Panel d is a merged image showing nuclear localization and panel e shows competition with the phospho-Tau (pT231) peptide. The images were captured at 20X magnification.

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  Immunohistochemistry paraffin embedded sections - Anti-Tau (phospho T231) antibody [RP23040023] (ab307987)

  Paraffin-embedded rat brain tissue stained for Tau (phospho T231) using ab307987 at 1/300 overnight at 4°C in a humidified chamber (right) compared to a negative control without primary antibody (left). To expose target proteins, antigen retrieval was performed using 10 mM sodium citrate (pH 6.0), microwaved for 8-15 min. Following antigen retrieval, tissues were blocked in 3% H2O2-methanol for 15 min at room temperature. Tissues were washed extensively in PBST and detection was performed using a HRP-conjugated secondary antibody followed by colorimetric detection using a DAB kit. Tissues were counterstained with hematoxylin and dehydrated with ethanol and xylene to prep for mounting.

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  Western blot - Anti-Tau (phospho T231) antibody [RP23040023] (ab307987)

  Western blot analysis of Tau (phospho T231) in total rat brain lysate using ab307987 at a dilution of 2.5µg/mL. To confirm specificity, competition was performed by preincubation with phosphopeptide to inhibit antibody binding (lane 2). Samples were detected using chemiluminescence (ECL).

  All lanes: Western blot - Anti-Tau (phospho T231) antibody [RP23040023] (ab307987) at 2.5 µg/mL

  Lane 1: total rat brain lysate without phosphopeptide

  Lane 2: total rat brain lysate with phosphopeptide

  Developed using the ECL technique.

  Observed band size: 56 kDa

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  Immunohistochemistry paraffin embedded sections - Anti-Tau (phospho T231) antibody [RP23040023] (ab307987)

  Immunohistochemical analysis of paraffin-embedded human glioma tissue labeling Phospho-Tau pThr231 with ab307987 at 1/100 dilution (right) and without ab307987 (left) followed by HRP-conjugated secondary antibody followed by colorimetric detection using a DAB kit. The sections were incubated with ab307987 overnight at 4degC. Counter stained with hematoxylin and dehydrated with ethanol and xylene to prep for mounting. Antigen retrieval was performed using 10mM sodium citrate (pH 6.0), microwaved for 8-15 min. Following antigen retrieval, tissues were blocked in 3% H2O2-methanol for 15 min at room temperature, washed with ddH2O and PBS. Tissues were washed extensively in PBST.