Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized mouse primary neural mix culture cells labelling Pan Trk with ab181560 at 1:100 dilution, followed by ab150077 AlexaFluor^®488 Goat anti-Rabbit secondary antibody at 1:1000 dilution (Green). Confocal scanning Z step was set as 0.3 µm followed by image processing with maximum Z projection. ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor^® 594) was used to counterstain tubulin at 1:200 dilution (Red). The Nuclear counterstain was DAPI (Blue).

Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% tritonX-100 permeabilized Neuro-2a (Mouse neuroblastoma cells) cells labeling Pan Trk with ab181560 at 1/250 dilution. Goat anti-rabbit IgG (Alexa Fluor^® 488) (ab150077) at 1/400 dilution was used as the secondary antibody (green). Confocal image showing cytoplasmic staining on Neuro-2a cells is shown. The nuclear counter stain is DAPI (blue). Tubulin is detected with ab7291 (Tubulin mouse mAb) at 1/500 and ab150120 (Alexa Fluor^® 594 Goat anti-Mouse secondary) at 1/500 dilution (red).

The negative controls are as follows;
1. ab181560 at 1/250 dilution followed by ab150120 (Goat anti mouse IgG (Alexa Fluor^® 594)) at 1/500 dilution.
2. ab7291 (anti-Tubulin mouse mAb) at 1/500 dilution followed by ab150077 (Goat anti rabbit IgG (Alexa Fluor^® 488)) at 1/400 dilution.

All lanes : Extraction Buffer (ab191560) at 1/10000 dilution

Lane 1 : Mouse brain lysates
Lane 2 : Rat brain lysates
Lane 3 : Mouse kidney lysates
Lane 4 : Mouse spleen lysates
Lane 5 : Rat kidney lysates
Lane 6 : Rat spleen lysates

Lysates/proteins at 10 µg per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG, (H+L),Peroxidase conjugated at 1/1000 dilution

Predicted band size: 92 kDa

TrkB is abundantly expressed in the central and peripheral nervous system. Mouse kidney, mouse spleen, rat kidney and rat spleen are used as negative control.
The 30KDa band is an intracellular fragment TrkB-ICD, and the 140KDa observed MW which is higher than the predicted one is due to the glycosylation modification.

Immunohistochemical analysis of paraffin-embedded Human astrocytoma tissue labeling Pan Trk with ab181560 at 1/500 dilution, followed by Anti-Rabbit HRP (ab97051) at 1/500 dilution. Astrocytoma cells show strong cytoplasmic staining. Counter stained with Hematoxylin. Antigen retrieval was perfomed using Tris-EDTA buffer, pH 9.0

Negative control: Using PBS instead of primary ab, secondary ab is prediluted HRP Polymer for Rabbit/Mouse IgG.

All lanes : Anti-Pan Trk antibody [EPR17341] (ab181560) at 1/10000 dilution

Lane 1 : Human fetal brain lysates
Lane 2 : Human fetal heart lysates
Lane 3 : Human fetal spleen lysates

Lysates/proteins at 20 µg per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG, (H+L),Peroxidase conjugated at 1/1000 dilution

Predicted band size: 92 kDa

TrkB is abundantly expressed in the central and peripheral nervous systems,

human fetal heart and human fetal spleen are used as negative controls.

The 30KDa band is an intracellular fragment TrkB-ICD, and the 140KDa observed MW which is higher than the predicted one is due to the glycosylation modification.

Blocking/dilution buffer: 5% NFDM/TBST.

Immunohistochemical analysis of paraffin-embedded Human cerebral cortex tissue labeling Pan Trk with ab181560 at 1/500 dilution, followed by Anti-Rabbit HRP (ab97051) at 1/500 dilution. Cytoplasmic staining is observed on neurons of human cerebral cortex. Counter stained with Hematoxylin. Antigen retrieval was perfomed using Tris-EDTA buffer, pH 9.0.

Negative control: Using PBS instead of primary ab, secondary ab is prediluted HRP Polymer for Rabbit/Mouse IgG.

Anti-Pan Trk antibody [EPR17341] (ab181560) at 1/10000 dilution + Human cerebellum lysates at 10 µg

Secondary
Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG at 1/1000 dilution

Predicted band size: 92 kDa

TrkB is abundantly expressed in the central and peripheral nervous system. The 30KDa band is an intracellular fragment TrkB-ICD, and the 140KDa observed MW which is higher than the predicted one is due to the glycosylation modification.

Blocking/dilution buffer: 5% NFDM/TBST.

Immunohistochemical analysis of paraffin-embedded Mouse cerebral cortex tissue labeling Pan Trk with ab181560 at 1/500 dilution, followed by Anti-Rabbit HRP (ab97051) at 1/500 dilution. Cytoplasmic staining is observed on neurons of mouse cerebral cortex. Counter stained with Hematoxylin. Antigen retrieval was perfomed using Tris-EDTA buffer, pH 9.0.
Negative control: Using PBS instead of primary ab, secondary ab is prediluted HRP Polymer for Rabbit/Mouse IgG.

Immunohistochemical analysis of paraffin-embedded Rat cerebral cortex tissue labeling Pan Trk with ab181560 at 1/500 dilution, followed by Anti-Rabbit HRP (ab97051) at 1/500 dilution. Cytoplasmic staining is observed on neurons of Rat cerebral cortex. Counter stained with Hematoxylin. Antigen retrieval was perfomed using Tris-EDTA buffer, pH 9.0.
Negative control: Using PBS instead of primary ab, secondary ab is prediluted HRP Polymer for Rabbit/Mouse IgG.

Immunohistochemical analysis of paraffin-embedded Human (Panel 1), Mouse (Panel 2) or Rat (Panel 3) liver tissue labeling Pan Trk with ab181560 at 1/500 dilution, followed by Anti-Rabbit HRP (ab97051) at 1/500 dilution. The staining is negative on normal Human liver. Counter stained with Hematoxylin. Antigen retrieval was perfomed using Tris-EDTA buffer, pH 9.0.
Negative control: Using PBS instead of primary ab, secondary ab is prediluted HRP Polymer for Rabbit/Mouse IgG.