Anti-NCAM1 antibody

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-NCAM1 antibody (AB95153)

IHC image of NCAM staining in Mouse foetal brain formalin fixed paraffin embedded tissue section, performed on a Leica BondTM system using the standard protocol B. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab95153, 1μg/ml, 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.

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.

• WB

Unknown

Western blot - Anti-NCAM1 antibody (AB95153)

This antibody recognises isoforms NCAM140 and NCAM180, at 140 and 180 kDa respectively. NCAM contains a number of potential glycosylation sites (SwissProt) which may explain its migration at a higher molecular weight than predicted. This banding pattern has been observed in the literature PMID : 21389209. Abcam recommends using milk as the blocking agent. Abcam welcomes customer feedback and would appreciate any comments regarding this product and the data presented above.

All lanes:

Western blot - Anti-NCAM1 antibody (ab95153) at 1 µg/mL

Lane 1:

Brain (Mouse) Tissue Lysate at 10 µg

Lane 2:

Forebrain (Mouse) Tissue Lysate at 10 µg

Lane 3:

Mouse Hippocampus Tissue Lysate at 10 µg

Lane 4:

Brain (Rat) Tissue Lysate at 10 µg

Lane 5:

Cerebellum Rat Tissue Lysate at 10 µg

Lane 6:

Rat Forebrain at 10 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) preadsorbed (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-preadsorbed-ab97080'>ab97080</a>) at 1/5000 dilution

Predicted band size: 94 kDa

Observed band size: 140 kDa,180 kDa

true

Exposure time: 30s