Anti-Tau (phospho S396) antibody [E178] - BSA and Azide free

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Tau (phospho S396) antibody [E178] - BSA and Azide free (AB218600)

This data was developed using the same antibody clone in a different buffer formulation (ab32057).

Immunohistochemistry analysis of paraffin-embedded human cerebrum tissue sections labeling Tau (phospho S396) with ab32057 at 1/4000 dilution (0.026 μg/mL). Goat Anti-Rabbit IgG H&L (HRP polymer) was used as the secondary antibody. Sections were counterstained with Hematoxylin. Antigen retrieval was heat mediated using ab93684 (Tris/EDTA buffer, pH 9.0).

Positive staining on human cerebrum without alkaline phosphatase treatment (image A). No staining on human cerebrum with alkaline phosphatase treatment (image B).
The section was incubated with ab32057 overnight at +4°C.

• IHC-P

Collaborator

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Tau (phospho S396) antibody [E178] - BSA and Azide free (AB218600)

This IHC data was generated using the same anti-Tau antibody clone, E178, in a different buffer formulation (cat# ab32057).

Immunohistochemistical detection of Tau antibody [E178] (ab32057) on formaldehyde-fixed paraffin-embedded human salivary gland sections. Antigen retrieval step : Heat mediated. Buffer Used : Citric acid pH6. Permeabilization : No. Blocking step : 1% BSA for 10 mins @ 21°C. ab32057 incubated at 1/1000 for 2 hours @ 21°C in TBS/BSA/azide. Secondary antibody : anti rabbit IgG conjugated to Biotin (1/200). NB : An interesting pattern of positivity that seems to be supported by the Human Protein Atlas. Coloured arrowheads in the submitted image indicate features : red for positive serous glands, blue for positive intra-lobular collecting ducts, black for negative mucous glands (there is a serous demilune around this acinus), yellow for intralobular collecting ducts, green for nerve tracks in the interlobular areas, blue for positive interlobular collecting ducts. There appears to be a population of positive nuclei but this may b

This image is courtesy of Carl Hobbs, King's College London, United Kingdom

• IHC-Fr

Lab

Immunohistochemistry (Frozen sections) - Anti-Tau (phospho S396) antibody [E178] - BSA and Azide free (AB218600)

This data was developed using the same antibody clone in a different buffer formulation (ab32057).

IHC image of Tau 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 ab32057, 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.

• IP

Unknown

Immunoprecipitation - Anti-Tau (phospho S396) antibody [E178] - BSA and Azide free (AB218600)

ab32057 (purified) at 1/20 dilution (0.5ug) immunoprecipitating Tau in Human fetal brain lysates.
Lane 1 : Human fetal brain lysates 10ug
Lane 2 (+) : ab32057 & Human fetal brain lysates
Lane 3 (-) : Rabbit monoclonal IgG (ab172730) instead of ab32057 in Human fetal brain lysates
For western blotting, VeriBlot for IP Detection Reagent (HRP) (ab131366) was used at 1/1000 dilution.
Blocking and diluting buffer : 5% NFDM/TBST.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab32057).

All lanes:

Immunoprecipitation - Anti-Tau (phospho S396) antibody [E178] (<a href='/en-us/products/primary-antibodies/tau-phospho-s396-antibody-e178-ab32057'>ab32057</a>)

Predicted band size: 78 kDa

false

• ELISA

Lab

ELISA - Anti-Tau (phospho S396) antibody [E178] - BSA and Azide free (AB218600)

This data was developed using the same antibody clone in a different buffer formulation (ab32057).

Indirect ELISA antigen dose-response curve using ab32057 at 1000-0 ng/mL. Antigen Human Tau (phospho S396) peptide, Human Tau non-phospho peptide at concentration of 1000 ng/mL. Alkaline Phosphatase-conjugated AffiniPure Goat Anti-Rabbit IgG H+L at 1/2500 dilution was used as the secondary antibody.

• WB

Lab

Western blot - Anti-Tau (phospho S396) antibody [E178] - BSA and Azide free (AB218600)

Blocking/Diluting buffer and concentration 5% NFDM/TBST

Tau assembles into oligomers as described in PMID : 28382304, 32692785 and 30120733.

This data was developed using the same antibody clone in a different buffer formulation (ab32057).

All lanes:

Western blot - Anti-Tau (phospho S396) antibody [E178] (<a href='/en-us/products/primary-antibodies/tau-phospho-s396-antibody-e178-ab32057'>ab32057</a>) at 1/1000 dilution

Lane 1:

Human brain lysates at 15 µg

Lane 2:

Human brain lysates and the membrane was incubated with alkaline phosphatase at 15 µg

Lane 3:

Human brain lysates and the membrane was incubated with lambda phosphatase at 15 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG (HRP) with minimal cross-reactivity with human IgG at 1/2000 dilution

Predicted band size: 78 kDa

Observed band size: 50-79 kDa

false

Exposure time: 60s

• AP

Lab

Affinity Purification - Anti-Tau (phospho S396) antibody [E178] - BSA and Azide free (AB218600)

Biotinylated Human Tau (pS396) [0.05 μg/ml] was loaded to SA biosensor on Fortebio RED96e Machine, then associate with recombinant Anti-Tau (phospho S396) antibody [E178] in serial concentration points [3.33, 1.67, 0.83, 0.42, 0.21 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 S396) antibody [E178] is 2.08E-11

This data was developed using the same antibody clone in a different buffer formulation (ab32057).

• WB

Lab

Western blot - Anti-Tau (phospho S396) antibody [E178] - BSA and Azide free (AB218600)

Blocking/Diluting buffer and concentration 5% NFDM/TBST

Tau assembles into oligomers as described in PMID : 28382304, 32692785 and 30120733.

This data was developed using the same antibody clone in a different buffer formulation (ab32057).

All lanes:

Western blot - Anti-Tau (phospho S396) antibody [E178] (<a href='/en-us/products/primary-antibodies/tau-phospho-s396-antibody-e178-ab32057'>ab32057</a>) at 1/1000 dilution

Lane 1:

Mouse brain lysates at 15 µg

Lane 2:

Mouse brain lysates and the membrane was incubated with alkaline phosphatase at 15 µg

Lane 3:

Mouse brain lysates and the membrane was incubated with lambda phosphatase at 15 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG (HRP) with minimal cross-reactivity with human IgG at 1/2000 dilution

Predicted band size: 78 kDa

Observed band size: 50-79 kDa

false

Exposure time: 10s

• Dot

Lab

Dot Blot - Anti-Tau (phospho S396) antibody [E178] - BSA and Azide free (AB218600)

This data was developed using the same antibody clone in a different buffer formulation (ab32057).

Dot blot analysis of Tau (phospho S396) phospho peptide (Lane 1) and Tau non-phospho peptide (Lane 2) labelling Tau (phospho S396) with ab32057 at a dilution of 1/1000. ab97051 (Peroxidase conjugated goat anti-rabbit IgG (H+L)) was used as the secondary antibody at a dilution of 1/100000.

Blocking and dilution buffer : 5% NFDM/TBST.

Exposure time : 3 minutes.