Immunohistochemical analysis of paraffin-embedded human glioma tissue labeling NCAM1 with ab237708 at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) ready to use. Positive staining on the human glioma is observed. Counter stained with hematoxylin.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) ready to use.

Heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0).

 

Immunohistochemical analysis of paraffin-embedded human cerebrum tissue labeling NCAM1 with ab237708 at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) ready to use. Positive staining on the human cerebrum is observed. Counter stained with hematoxylin.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) ready to use.

Heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0).

 

Immunohistochemical analysis of paraffin-embedded human pancreas tissue labeling NCAM1 with ab237708 at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) ready to use. Positive staining on islet in the human pancreas is observed. Counter stained with hematoxylin.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) ready to use.

Heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0).

 

Immunohistochemical analysis of paraffin-embedded mouse cerebrum tissue labeling NCAM1 with ab237708 at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) ready to use. Positive staining on islet in the mouse cerebrum is observed. Counter stained with hematoxylin.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) ready to use.

Heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0).

 

Immunohistochemical analysis of paraffin-embedded rat cerebrum tissue labeling NCAM1 with ab237708 at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) ready to use. Positive staining on islet in the rat cerebrum is observed. Counter stained with hematoxylin.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) ready to use.

Heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0).

Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized Neuro-2a (mouse neuroblastoma cell line) cells labeling NCAM1 with ab237708 at 1/50 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) (ab150077) secondary antibody at 1/1000 dilution (green).Confocal image showing membranous staining in Neuro-2a cells. The nuclear counter stain is DAPI (blue). Tubulin is detected with Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor^® 594) (ab195889) at 1/200 dilution (red).

PCS only control: Used PBS instead of primary antibody, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) (ab150077) secondary antibody at 1/1000 dilution.

Negative control: L-929 (PMID:9696812).

NCAM1 was immunoprecipitated from 0.35 mg of SH-SY5Y (Human neuroblastoma cell line from bone marrow) whole cell lysate with ab237708 at 1/20 dilution. Western blot was performed from the immunoprecipitate using ab237708 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used as secondary antibody at 1/5000 dilution.

Lane 1: SH-SY5Y whole cell lysate 10 μg (Input).
Lane 2: ab237708 IP in SH-SY5Y whole cell lysate.
Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab237708 in SH-SY5Y whole cell lysate.

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

All lanes : Anti-NCAM1 antibody [CAL53] (ab237708) at 1/1000 dilution

Lane 1 : Human brain lysate
Lane 2 : Human cerebellum lysate
Lane 3 : Neuro-2a (mouse neuroblastoma cell line0 whole cell lysate
Lane 4 : Mouse cerebellum lysate
Lane 5 : Rat brain lysate

Lysates/proteins at 20 µg per lane.

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

Predicted band size: 94 kDa
Observed band size: 140-180 kDa why is the actual band size different from the predicted?


Exposure time: 26 seconds

Blocking and dilution buffer: 5% NFDM/TBST.

Formalin-fixed, paraffin-embedded human pancreatic carcinoma tissue stained for NCAM1 using ab237708 at 0.25 µg/ml in immunohistochemical analysis.