Anti-L1CAM antibody [EPR18998]

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-L1CAM antibody [EPR18998] (AB182407)

Immunohistochemical analysis of paraffin-embedded Human brain tissue labeling L1CAM with ab182407 at 1/250 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Cytoplasm staining on the neurons in Human brain 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) (ab97051) at 1/500 dilution.

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

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-L1CAM antibody [EPR18998] (AB182407)

Immunohistochemical analysis of paraffin-embedded Human kidney tissue labeling L1CAM with ab182407 at 1/250 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Membrane and cytoplasm staining on the Human kidney 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) (ab97051) at 1/500 dilution.

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

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-L1CAM antibody [EPR18998] (AB182407)

Immunohistochemical analysis of paraffin-embedded Human liver tissue labeling L1CAM with ab182407 at 1/250 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Negative staining on the Human liver is observed. Counter stained with Hematoxylin.

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

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-L1CAM antibody [EPR18998] (AB182407)

Immunohistochemical analysis of paraffin-embedded Human breast cancer tissue labeling L1CAM with ab182407 at 1/250 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Membrane and cytoplasm staining on Human breast cancer 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) (ab97051) at 1/500 dilution.

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

• WB

Supplier Data

Western blot - Anti-L1CAM antibody [EPR18998] (AB182407)

Blocking/Dilution buffer : 5% NFDM/TBST.

Exposure time : Lane 1 and 2 : 3minutes; Lane 3 : 2 seconds; Lane 4 : 10 seconds.

Observed MW : 200-220, 85 kDa

ab182407 binds to the full length L1CAM and the 85kDa fragment. Plasmin cleaves L1CAM at the FN3 repeat to produce 140 kDa and 85 kDa fragments (PMID : 7542658; PMID : 18332088; PMID : 20840789). The 85 kDa fragment is where the immunogen is located.

All lanes:

Western blot - Anti-L1CAM antibody [EPR18998] (ab182407) at 1/5000 dilution

Lane 1:

Human glioma lysate at 20 µg

Lane 2:

A-375 (Human malignant melanoma cell line) whole cell lysate at 20 µg

Lane 3:

Human cerebellum lysate at 20 µg

Lane 4:

Human hippocampus lysate at 20 µg

Secondary

All lanes:

Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG at 1/10000 dilution

Predicted band size: 140 kDa

false

• WB

Supplier Data

Western blot - Anti-L1CAM antibody [EPR18998] (AB182407)

Blocking/Dilution buffer : 5% NFDM/TBST.

Exposure time : Lane 1 : 1 second; Lane 2 : 3 minutes.

Observed MW : 200-220, 85 kDa

ab182407 binds to the full length L1CAM and the 85kDa fragment. Plasmin cleaves L1CAM at the FN3 repeat to produce 140 kDa and 85 kDa fragments (PMID : 7542658; PMID : 18332088; PMID : 20840789). The 85 kDa fragment is where the immunogen is located.

All lanes:

Western blot - Anti-L1CAM antibody [EPR18998] (ab182407) at 1/1000 dilution

Lane 1:

Human fetal brain lysate at 10 µg

Lane 2:

Human fetal kidney lysate at 10 µg

Secondary

All lanes:

Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG at 1/10000 dilution

Predicted band size: 140 kDa

false

• WB

Unknown

Western blot - Anti-L1CAM antibody [EPR18998] (AB182407)

Lanes 1-2 : Merged signal (red and green). Green - ab182407 observed at 220 kDa. Red - loading control, ab130007 observed at 125 kDa.

ab182407 was shown to react with L1CAM in wild-type HeLa cells in Western blot. Loss of signal was observed when knockout sample ab263786 was used. Wild-type and L1CAM knockout samples were subjected to SDS-PAGE. ab182407 and Anti-Vinculin antibody [VIN-54] (ab130007) were incubated overnight at 4°C at 1 in 5000 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.

All lanes:

Western blot - Anti-L1CAM antibody [EPR18998] (ab182407) at 1/5000 dilution

Lane 1:

Wild-type HeLa cell lysate at 20 µg

Lane 2:

L1CAM knockout HeLa cell lysate at 20 µg

Lane 2:

Western blot - Human L1CAM knockout HeLa cell line (<a href='/en-us/products/cell-lines/human-l1cam-knockout-hela-cell-line-ab255401'>ab255401</a>)

Secondary

All lanes:

Western blot - Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-irdye-800cw-preadsorbed-ab216773'>ab216773</a>) at 1/20000 dilution

Predicted band size: 140 kDa

Observed band size: 124 kDa,220 kDa

false