Immunohistochemical analysis of paraffin-embedded Human tonsil tissue labeling S100 with ab183979 at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Positive on germinal center of Human tonsil. Counter stained with Hematoxylin.

Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is ab97051 at 1/500 dilution.

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

C2C12 (mouse muscle) cells were fixed with 4% paraformaldehyde and incubated with the primary antibody (ab183979; red line) diluted 1:60 (1µg).  The secondary antibody used was Goat anti-rabbit IgG (Alexa Fluor^® 488) at 1:500 dilution.  The isotype control (black line) antibody was Rabbit monoclonal IgG (ab172730).  An unlabelled control without incubation with primary antibody and secondary antibody was also perfomed (blue line).

Immunohistochemical analysis of paraffin-embedded Human cerebrum tissue labeling S100 with ab183979 at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Positive staining on glial cells and negative on neuron cells of Human cerebrum was observed. Counter stained with Hematoxylin.

Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is ab97051 at 1/500 dilution.

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

Immunohistochemical analysis of paraffin-embedded Human kidney tissue labeling S100 with ab183979 at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Part of kidney tubules were strongly staining. Counter stained with Hematoxylin.

Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is ab97051 at 1/500 dilution.

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

Immunohistochemical analysis of 4% paraformaldehyde-fixed, 0.2% Triton X-100 permeabilized frozen Mouse brain tissue labeling S100 with ab183979 at 1/500 dilution, followed by Goat Anti-Rabbit IgG (Alexa Fluor^® 488) (ab150077) secondary antibody at 1/1000 dilution (green). The result showed positive staining on glial cells of mouse cortex. The nuclear counterstain is DAPI (blue).

Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is ab150077 secondary antibody at 1/1000 dilution.

Immunohistochemical analysis of 4% paraformaldehyde-fixed, 0.2% Triton X-100 permeabilized frozen Mouse kidney tissue labeling S100 with ab183979 at 1/500 dilution, followed by Goat Anti-Rabbit IgG (Alexa Fluor^® 488) (ab150077) secondary antibody at 1/1000 dilution (green). The result showed positive staining on mouse kidney. The nuclear counterstain is DAPI (blue).

Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is ab150077 secondary antibody at 1/1000 dilution.

Immunohistochemical analysis of 4% paraformaldehyde-fixed, 0.2% Triton X-100 permeabilized frozen Mouse spleen tissue labeling S100 with ab183979 at 1/500 dilution, followed by Goat Anti-Rabbit IgG (Alexa Fluor^® 488) (ab150077) secondary antibody at 1/1000 dilution (green). The result showed positive staining on mouse spleen. The nuclear counterstain is DAPI (blue).

Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is ab150077 secondary antibody at 1/1000 dilution.

All lanes : Anti-S100 alpha antibody [EPR19013] (ab183979) at 1/2000 dilution

Lane 1 : Human skeletal muscle lysate
Lane 2 : Mouse cerebral cortex lysate
Lane 3 : Mouse liver lysate

Lysates/proteins at 20 µg per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG Peroxidase Conjugate, specific to the non-reduced form of IgG at 1/10000 dilution

Predicted band size: 11 kDa
Observed band size: 11 kDa


Exposure time: 3 minutes

Blocking/Dilution buffer: 5% NFDM/TBST.

Note that high molecular weight bands were detected in mouse tissue lysates.

Anti-S100 alpha antibody [EPR19013] (ab183979) at 1/2000 dilution + RAW 264.7 (Mouse macrophage cell line transformed with Abelson murine leukemia virus) whole cell lysate at 10 µg

Secondary
Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

Predicted band size: 11 kDa
Observed band size: 11 kDa


Exposure time: 3 minutes

Blocking/Dilution buffer: 5% NFDM/TBST.

All lanes : Anti-S100 alpha antibody [EPR19013] (ab183979) at 1/1000 dilution

Lane 1 : Mouse muscle lysate
Lane 2 : Mouse heart lysate

Lysates/proteins at 10 µg per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

Predicted band size: 11 kDa
Observed band size: 11 kDa


Exposure time: 3 minutes

Blocking/Dilution buffer: 5% NFDM/TBST.

Note that high molecular weight bands were detected in mouse tissue lysates.

S100 was immunoprecipitated from RAW 264.7 (Mouse macrophage cell line transformed with Abelson murine leukemia virus) whole cell lysate with ab183979 at 1/40 dilution. Western blot was performed from the immunoprecipitate using ab183979 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/10000 dilution.

Lane 1: RAW 264.7 whole cell lysate, 10µg (Input).

Lane 2: ab183979 IP in RAW 264.7 whole cell lysate.

Lane 3: Rabbit IgG, monoclonal [EPR25A]  - Isotype Control (ab172730) instead of ab183979 in RAW 264.7 whole cell lysate.

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

Exposure time: 30 seconds.