Recombinant Anti-NCAM1 antibody [EP2567Y] (ab75813)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human cerebrum tissue sections labeling NCAM1 with Purified ab75813 at 1/10000 dilution (0.02 µg/ml). Heat mediated antigen retrieval was performed using Heat mediated antigen retrieval using Bond™ Epitope Retrieval Solution 2 (pH 9.0) . Tissue was counterstained with Hematoxylin. Followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101) secondary antibody. PBS instead of the primary antibody was used as the negative control.

Immunocytochemistry analysis of SH-SY5Y (Human neuroblastoma epithelial cell) cells labeling NCAM1 with Purified ab75813 at 1:50 dilution (3.1 µg/ml). Cells were fixed in 4% Paraformaldehyde and permeabilized with 0.1% tritonX-100. Cells were counterstained with Ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) 1:200 (2.5 µg/ml). Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) was used as the secondary antibody at 1:1000 (2 µg/ml) dilution. DAPI (blue) was used as nuclear counterstain. PBS instead of the primary antibody was used as the secondary antibody only control.

Flow Cytometry analysis of SH-SY5Y (Human neuroblastoma epithelial cell) cells labelling NCAM1 with Purified ab75813 at 1:20 dilution (10 µg/ml) (Red). Cells were fixed with 4% Paraformaldehyde and permeabilised with 90% Methanol. A Goat anti rabbit IgG (Alexa Fluor® 488, ab150081) secondary antibody was used at 1:2000. Isotype control - Rabbit monoclonal IgG (Black). Unlabelled control - Cell without incubation with primary antibody and secondary antibody (Blue).

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human astrocytoma tissue sections labeling NCAM1 with Purified ab75813 at 1:10000 dilution (0.0156 µg/ml). Heat mediated antigen retrieval was performed using Heat mediated antigen retrieval using Bond™ Epitope Retrieval Solution 2 (pH 9.0) . Tissue was counterstained with Hematoxylin. Rabbit specific IHC polymer detection kit HRP/DAB (ab209101) secondary antibody was used at 1:0 dilution. PBS instead of the primary antibody was used as the negative control. 

NCAM1 was immunoprecipitated from 0.35 mg SH-SY5Y (Human neuroblastoma epithelial cell) whole cell lysate 10 µg with 75813 at 1/50 dilution (2µg). VeriBlot for IP Detection Reagent (HRP)(ab131366) was used at 1/5000 dilution.

Lane 1: SH-SY5Y (Human neuroblastoma epithelial cell) whole cell lysate 10 µg

Lane 2: ab75813 IP in SH-SY5Y whole cell lysate

Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab75813 in SH-SY5Y whole cell lysate

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