Anti-Tau (phospho T231) 抗体 [EPR2488]

• IHC-P

Collaborator

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Tau (phospho T231) antibody [EPR2488] (AB151559)

IHC image of Tau (phospho T231) 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 ab151559 at 1/2000 dilution for 2 hours at 21°C. 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.

Image courtesy of Carl Hobbs, Kings College London.

• IHC-Fr

Lab

Immunohistochemistry (Frozen sections) - Anti-Tau (phospho T231) antibody [EPR2488] (AB151559)

IHC image of Tau (phospho T231) staining in a section of frozen normal human Azheimer 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 ab151559, 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.

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Tau (phospho T231) antibody [EPR2488] (AB151559)

IHC image of Tau (phospho T231) staining in human Alzheimer hippocampus formalin fixed paraffin embedded tissue section, performed on a Leica Bond system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6, epitope retrieval solution 1) for 20 minutes. The section was then incubated with unpurified ab151559, 1/200 dilution, for 15 minutes 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.

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.

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Tau (phospho T231) antibody [EPR2488] (AB151559)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Human cerebrum tissue sections labeling Tau with Purified ab151559 at 1 : 500 dilution (0.24 μg/ml). Heat mediated antigen retrieval was performed using citrate (pH 6.0)

• I-ELISA

Lab

Indirect ELISA - Anti-Tau (phospho T231) antibody [EPR2488] (AB151559)

Indirect ELISA analysis of ab151559 at 1000-0 ng/ml. The Secondary antibody used was Alkaline Phosphatase-conjugated AffiniPure Goat Anti-Rabbit IgG (H+L) at 1 : 2500 dilution.

Antigen : Human Tau (phospho T231) peptide, Human Tau non-phospho peptide

Antigen concentration : 1000 ng/ml.

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Tau (phospho T231) antibody [EPR2488] (AB151559)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Mouse cerebrum tissue sections labeling Tau or Primary ab team with Purified ab151559 at 1 : 500 dilution (0.24 μg/ml). Heat mediated antigen retrieval was performed using citrate (pH 6.0)

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Tau (phospho T231) antibody [EPR2488] (AB151559)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Rat cerebrum tissue sections labeling Tau with Purified ab151559 at 1 : 500 dilution (0.24 μg/ml). Heat mediated antigen retrieval was performed using citrate (pH 6.0)

• IHC-Fr

Unknown

Immunohistochemistry (Frozen sections) - Anti-Tau (phospho T231) antibody [EPR2488] (AB151559)

Immunohistochemistry (Frozen sections) analysis of mouse cerebrum tissue sections labeling Tau with Purified ab151559 at 1/50 (1.5 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.

• IP

Lab

Immunoprecipitation - Anti-Tau (phospho T231) antibody [EPR2488] (AB151559)

ab151559 (purified) at 1 : 20 dilution (0.6μg) immunoprecipitating Tau in Rat cerebral cortex lysate.
Lane 1 (input) : Rat cerebral cortex lysate 10μg
Lane 2 (+) : ab151559 & Rat cerebral cortex lysate
Lane 3 (-) : Rabbit monoclonal IgG (ab172730) instead of ab151559 inRat cerebral cortex lysate
For western blotting, VeriBlot for IP Detection Reagent (HRP) (ab131366) was used for detection at 1 : 1000 dilution.
Blocking and diluting buffer : 5% NFDM/TBST.

All lanes:

Immunoprecipitation - Anti-Tau (phospho T231) antibody [EPR2488] (ab151559)

Predicted band size: 78 kDa

Observed band size: 50-70 kDa

false

• WB

Lab

Western blot - Anti-Tau (phospho T231) antibody [EPR2488] (AB151559)

Blocking and dilution buffer : 5% NFDM/TBST.

All lanes:

Western blot - Anti-Tau (phospho T231) antibody [EPR2488] (ab151559) at 1/1000 dilution

Lane 1:

Human hippocampus tissue lysate at 15 µg

Lane 2:

Human hippocampus tissue lysate incubated with phosphatase at 15 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/20000 dilution

Predicted band size: 78 kDa

Observed band size: 50-70 kDa

false

Exposure time: 1s

• WB

Lab

Western blot - Anti-Tau (phospho T231) antibody [EPR2488] (AB151559)

Blocking and dilution buffer : 5% NFDM/TBST.

Western blot for ab205606 is positioned on the right-hand side and serves as a control for the detection of the flag tag.

All lanes:

Western blot - Anti-Tau (phospho T231) antibody [EPR2488] (ab151559) at 1/1000 dilution

Lane 1:

293T cells transfected with an empty vector containing a flag tag whole cell lysate at 20 µg

Lane 2:

293T cells transfected with a human Tau expression vector containing a flag whole cell lysate at 20 µg

Lane 3:

293T cells transfected with a human MAP2 expression vector containing a flag whole cell lysate at 20 µg

Lane 4:

293T cells transfected with a human MAP4 expression vector containing a flag whole cell lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/20000 dilution

Observed band size: 50-100 kDa

false

Exposure time: 1s

• AP

Lab

Affinity Purification - Anti-Tau (phospho T231) antibody [EPR2488] (AB151559)

Biotinylated Human Tau (pT231) [0.3125 μg/ml] was loaded to SA biosensor on Fortebio RED96e Machine, then associate with recombinant Anti-Tau (phospho T231) antibody [EPR2488] in serial concentration points [4.17, 2.08, 1.04, 0.52, 0.26 nM/mL] by 2-fold dilution, next to dissociate in blank testing buffer [0.1% BSA in PBST (0.05%Tween-20)]. Calculated signals had already subtracted blank control, curve fitting using 1 : 1 binding model. Non-phospho Tau protein's association and dissociation were also showed in graph. KD(M) value of Anti-Tau (phospho T231) antibody [EPR2488] is 1.26E-11

• WB

Lab

Western blot - Anti-Tau (phospho T231) antibody [EPR2488] (AB151559)

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

All lanes:

Western blot - Anti-Tau (phospho T231) antibody [EPR2488] (ab151559) at 1/5000 dilution

Lane 1:

Rat cerebral cortex lysates at 15 µg

Lane 2:

Rat cerebral cortex lysates. Then the membrane was incubated with phosphatase. at 15 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/20000 dilution

Predicted band size: 78 kDa

Observed band size: 50-70 kDa

false

• WB

Lab

Western blot - Anti-Tau (phospho T231) antibody [EPR2488] (AB151559)

Blocking and dilution buffer : 5% NFDM/TBST.

All lanes:

Anti-cardiac Troponin I antibody (ab1000) at 1/1000 dilution

Lane 1:

C57 (cerebral cortex) whole cell lysate at 15 µg

Lane 2:

C57 (cerebral cortex) whole cell lysate incubated with phosphatase at 15 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/20000 dilution

Predicted band size: 78 kDa

Observed band size: 50-70 kDa

false

Exposure time: 1s

• WB

Unknown

Western blot - Anti-Tau (phospho T231) antibody [EPR2488] (AB151559)

All lanes:

Western blot - Anti-Tau (phospho T231) antibody [EPR2488] (ab151559) at 1/1000 dilution

Lane 1:

SH-SY5Y (human neuroblastoma cell line from bone marrow) cell lysate, untreated at 10 µg

Lane 2:

SH-SY5Y cell lysate, treated with sorbitol at 10 µg

Predicted band size: 78 kDa

false