Anti-L1CAM antibody [EPR18750]

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-L1CAM antibody [EPR18750] (AB208155)

Immunohistochemical analysis of paraffin-embedded Human stomach cancer tissue labeling L1CAM with ab208155 at 1/500 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

Membrane staining on the tumor cells of Human stomach cancer is observed.
L1CAM specific staining most abundant on nervous system, distal kidney tubules, and tumor cells. [PMID : 16867862, PMID : 20044598].

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 [EPR18750] (AB208155)

Immunohistochemical analysis of paraffin-embedded Human liver tissue labeling L1CAM with ab208155 at 1/500 dilution followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

Negative staining on the Human liver.

L1CAM specific staining most abundant on nervous system distal kidney tubules and tumor cells. [PMID : 16867862 PMID : 20044598].

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 [EPR18750] (AB208155)

Immunohistochemical analysis of paraffin-embedded Human kidney tissue labeling L1CAM with ab208155 at 1/500 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

Membrane staining on a part of Human kidney tubules is observed.
L1CAM specific staining most abundant on nervous system, distal kidney tubules, and tumor cells. [PMID : 16867862, PMID : 20044598].

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.

• IP

Supplier Data

Immunoprecipitation - Anti-L1CAM antibody [EPR18750] (AB208155)

L1CAM was immunoprecipitated from 1mg of Human cerebellum lysate with ab208155 at 1/40 dilution. Western blot was performed from the immunoprecipitate using ab208155 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/10000 dilution.
Lane 1 : Human cerebellum lysate 10μg (Input).
Lane 2 : ab208155 IP in Human cerebellum lysate.
Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab208155 in Human cerebellum lysate.
Blocking and dilution buffer and concentration : 5% NFDM/TBST.
Exposure time : 3 minutes.

All lanes:

Immunoprecipitation - Anti-L1CAM antibody [EPR18750] (ab208155)

Predicted band size: 140 kDa

false

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-L1CAM antibody [EPR18750] (AB208155)

Immunohistochemical analysis of paraffin-embedded Mouse testis tissue labeling L1CAM with ab208155 at 1/500 dilution followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

Negative staining on the mouse testis.
L1CAM specific staining most abundant on nervous system distal kidney tubules and tumor cells. [PMID : 16867862 PMID : 20044598].

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 [EPR18750] (AB208155)

Immunohistochemical analysis of paraffin-embedded Rat skeletal muscle tissue labeling L1CAM with ab208155 at 1/500 dilution followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

Negative staining on the rat skeletal muscle.
L1CAM specific staining most abundant on nervous system distal kidney tubules and tumor cells. [PMID : 16867862 PMID : 20044598].

Counter stained with Hematoxylin.

Secondary antibody only control : Used PBS instead of primary antibody secondary antibody is 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 [EPR18750] (AB208155)

Immunohistochemical analysis of paraffin-embedded Mouse cerebrum tissue labeling L1CAM with ab208155 at 1/500 dilution followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

Cytoplasm staining on the mouse cerebrum is observed.
L1CAM specific staining most abundant on nervous system distal kidney tubules and tumor cells. [PMID : 16867862 PMID : 20044598].

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 [EPR18750] (AB208155)

Immunohistochemical analysis of paraffin-embedded Rat colon tissue labeling L1CAM with ab208155 at 1/500 dilution followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

Cytoplasm staining on the nerve tract of the rat colon is observed.
L1CAM specific staining most abundant on nervous system distal kidney tubules and tumor cells. [PMID : 16867862 PMID : 20044598].

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 [EPR18750] (AB208155)

Immunohistochemical analysis of paraffin-embedded Mouse colon tissue labeling L1CAM with ab208155 at 1/500 dilution followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

Mainly membrane staining on the nerve tract of mouse colon is observed.
L1CAM specific staining most abundant on nervous system distal kidney tubules and tumor cells. [PMID : 16867862 PMID : 20044598].

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 [EPR18750] (AB208155)

Immunohistochemical analysis of paraffin-embedded Rat cerebellum tissue labeling L1CAM with ab208155 at 1/500 dilution followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

Cytoplasm staining on the molecular layer of the rat cerebellar cortex is observed.
L1CAM specific staining most abundant on nervous system distal kidney tubules and tumor cells. [PMID : 16867862 PMID : 20044598].

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.

• IP

Supplier Data

Immunoprecipitation - Anti-L1CAM antibody [EPR18750] (AB208155)

L1CAM was immunoprecipitated from 1mg of Rat brain whole cell lysate with ab208155 at 1/40 dilution. Western blot was performed from the immunoprecipitate using ab208155 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/10000 dilution.
Lane 1 : Rat brain whole cell lysate 10μg (Input).
Lane 2 : ab208155 IP in Rat brain whole cell lysate.
Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab208155 in Rat brain whole cell lysate.
Blocking and dilution buffer and concentration : 5% NFDM/TBST.
Exposure time : 3 minutes.

All lanes:

Immunoprecipitation - Anti-L1CAM antibody [EPR18750] (ab208155)

Predicted band size: 140 kDa

false

• WB

Supplier Data

Western blot - Anti-L1CAM antibody [EPR18750] (AB208155)

Blocking/Dilution buffer : 5% NFDM/TBST.

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

All lanes:

Western blot - Anti-L1CAM antibody [EPR18750] (ab208155) at 1/1000 dilution

Lane 1:

Mouse cerebellum lysate at 10 µg

Lane 2:

Mouse brain lysate at 10 µg

Lane 3:

A-375 (Human malignant melanoma cell line) whole cell lysate at 10 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/100000 dilution

Predicted band size: 140 kDa

Observed band size: 140 kDa,200-220 kDa

false

Exposure time: 3min

• WB

Lab

Western blot - Anti-L1CAM antibody [EPR18750] (AB208155)

False colour image of Western blot : Anti-L1CAM antibody [EPR18750] staining at 1/1000 dilution, shown in green; Mouse anti-Alpha Tubulin [DM1A] (ab7291) loading control staining at 1/20000 dilution, shown in red. In Western blot, ab208155 was shown to bind specifically to L1CAM. A band was observed at 220 kDa in wild-type HeLa cell lysates with no signal observed at this size in L1CAM knockout cell line ab273836 (knockout cell lysate ab273790). The band observed in the knockout lysate lane below 220 kDa is likely to represent a truncated form of L1CAM. This has not been investigated further and the functional properties of the gene product have not been determined. To generate this image, wild-type and L1CAM knockout HeLa cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3 % milk in TBS-0.1 % Tween^® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit IgG H&L (IRDye^® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preabsorbed (ab216776) at 1/20000 dilution.

All lanes:

Western blot - Anti-L1CAM antibody [EPR18750] (ab208155) at 1/1000 dilution

All lanes:

Wild-type HeLa cell lysate at 20 µg

Predicted band size: 140 kDa

Observed band size: 220 kDa

false

• WB

Unknown

Western blot - Anti-L1CAM antibody [EPR18750] (AB208155)

Lanes 1-2 : Merged signal (red and green). Green - ab208155 observed at 220 kDa. Red - loading control, ab130007 observed at 125 kDa.

ab208155 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. ab208155 and Anti-Vinculin antibody [VIN-54] (ab130007) were incubated overnight at 4°C at 1 in 1000 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 [EPR18750] (ab208155) at 1/1000 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

• WB

Supplier Data

Western blot - Anti-L1CAM antibody [EPR18750] (AB208155)

Blocking/Dilution buffer : 5% NFDM/TBST.

Exposure time : Lane 1 and 2 : 30seconds; Lane 3 : 3 minutes.

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

All lanes:

Western blot - Anti-L1CAM antibody [EPR18750] (ab208155) at 1/2000 dilution

Lane 1:

Human fetal brain lysate at 20 µg

Lane 2:

Human cerebellum lysate at 20 µg

Lane 3:

HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate at 20 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG Peroxidase Conjugate, specific to the non-reduced form of IgG at 1/10000 dilution

Predicted band size: 140 kDa

Observed band size: 140 kDa,200-220 kDa

false

• WB

Supplier Data

Western blot - Anti-L1CAM antibody [EPR18750] (AB208155)

Blocking/Dilution buffer : 5% NFDM/TBST.

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

All lanes:

Western blot - Anti-L1CAM antibody [EPR18750] (ab208155) at 1/2000 dilution

Lane 1:

Rat brain lysate at 20 µg

Lane 2:

Rat cerebellum lysate at 20 µg

Lane 3:

Rat hippocampus lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/100000 dilution

Predicted band size: 140 kDa

Observed band size: 140 kDa,200-220 kDa

false

Exposure time: 3min