Anti-L1CAM antibody [EPR18998] (ab182407)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR18998] to L1CAM
- Suitable for: WB, IHC-P
- Knockout validated
- Reacts with: Human
Overview
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Product name
Anti-L1CAM antibody [EPR18998]
See all L1CAM primary antibodies -
Description
Rabbit monoclonal [EPR18998] to L1CAM -
Host species
Rabbit -
Tested Applications & Species
See all applications and species dataApplication Species IHC-P HumanWB Human -
Immunogen
Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.
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Positive control
- WB: Human fetal brain, fetal kidney, glioma, cerebellum and hippocampus lysates; A-375 and HeLa whole cell lysate. IHC-P: Human kidney, brain and breast cancer tissues.
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General notes
This product is a recombinant monoclonal antibody, which offers several advantages including:
- - High batch-to-batch consistency and reproducibility
- - Improved sensitivity and specificity
- - Long-term security of supply
- - Animal-free production
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle. -
Storage buffer
pH: 7.2
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA -
Concentration information loading...
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Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR18998 -
Isotype
IgG -
Research areas
Images
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All lanes : Anti-L1CAM antibody [EPR18998] (ab182407) at 1/5000 dilution
Lane 1 : Wild-type HeLa cell lysate
Lane 2 : L1CAM knockout HeLa cell lysate
Lysates/proteins at 20 µg per lane.
Predicted band size: 140 kDaLanes 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. Loss of signal was observed when knockout cell line ab255401 (knockout cell lysate 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.
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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.
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All lanes : Anti-L1CAM antibody [EPR18998] (ab182407) at 1/1000 dilution
Lane 1 : Human fetal brain lysate
Lane 2 : Human fetal kidney lysate
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG at 1/10000 dilution
Predicted band size: 140 kDaBlocking/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.
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All lanes : Anti-L1CAM antibody [EPR18998] (ab182407) at 1/5000 dilution
Lane 1 : Human glioma lysate
Lane 2 : A-375 (Human malignant melanoma cell line) whole cell lysate
Lane 3 : Human cerebellum lysate
Lane 4 : Human hippocampus lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG at 1/10000 dilution
Predicted band size: 140 kDaBlocking/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.
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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.
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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.
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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.
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