Mouse Monoclonal L1CAM antibody. Suitable for IHC-P, ICC, WB and reacts with Rat, Human, Mouse samples. Cited in 58 publications.
IgG2b
Mouse
Preservative: 0.02% Sodium azide
Constituents: PBS, 6.97% L-Arginine
Liquid
Monoclonal
IHC-P | ICC | WB | |
---|---|---|---|
Human | Tested | Tested | Tested |
Mouse | Expected | Expected | Tested |
Rat | Tested | Tested | Tested |
Species | Dilution info | Notes |
---|---|---|
Species Rat | Dilution info 0.10000-0.50000 µg/mL | Notes Unsuitable for IHC-P in mouse tissue from in-house testing. Perform heat-mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol. |
Species Human | Dilution info 0.10000-0.50000 µg/mL | Notes Unsuitable for IHC-P in mouse tissue from in-house testing. Perform heat-mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol. |
Species | Dilution info | Notes |
---|---|---|
Species Mouse | Dilution info Use at an assay dependent concentration. | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Rat | Dilution info 1 µg/mL | Notes - |
Species Human | Dilution info 1 µg/mL | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Mouse | Dilution info Use at an assay dependent concentration. | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Mouse | Dilution info 1 µg/mL | Notes Cleavage products observed at 60-80 kDa. |
Species Rat | Dilution info 1 µg/mL | Notes Cleavage products observed at 60-80 kDa. |
Species Human | Dilution info 1 µg/mL | Notes Cleavage products observed at 60-80 kDa. |
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Neural cell adhesion molecule involved in the dynamics of cell adhesion and in the generation of transmembrane signals at tyrosine kinase receptors. During brain development, critical in multiple processes, including neuronal migration, axonal growth and fasciculation, and synaptogenesis. In the mature brain, plays a role in the dynamics of neuronal structure and function, including synaptic plasticity.
CAML1, MIC5, L1CAM, CAML1, MIC5, Neural cell adhesion molecule L1, N-CAM-L1, NCAM-L1
Mouse Monoclonal L1CAM antibody. Suitable for IHC-P, ICC, WB and reacts with Rat, Human, Mouse samples. Cited in 58 publications.
IgG2b
Mouse
Preservative: 0.02% Sodium azide
Constituents: PBS, 6.97% L-Arginine
Liquid
Monoclonal
2C2
Affinity purification Protein G
This clone may cross-react with CHL1 (PMID: 20215505).
Purified from TCS.
Blue Ice
1-2 weeks
+4°C
-20°C
Upon delivery aliquot
Avoid freeze / thaw cycle
L1CAM can be detected between 200-220 kD. In brain samples it is typically seen at ~ 200 kD. When the protein is overexpressed in vitro it is often detected as a doublet with bands at 200 and 220 kD. The unglycosylated, unprocessed L1CAM is ~ 140-150 kDa. The protein has 21 putative N-glycosylation sites on the extracellular portion of the protein which, when they are all glycosylated, results in a detected MW of 200-220 kD depending upon how many residues are actually glycosylated. L1CAM can be cleaved by the metalloprotease ADAM10 resulting in fragments of 180 kD and 40 kD. L1CAM can also be cleaved by plasmin resulting in fragments of 140 kD and 80 kD. In theory, therefore, one could detect bands at ~220, 200, 180, 140, 80 and 40 kD.
This product was changed from ascites to tissue culture supernatant on 08/Jul/2019. Lot numbers higher than GR3248431 are from tissue culture supernatant. Please note that the dilutions may need to be adjusted accordingly. If you have any questions, please do not hesitate to contact our scientific support team.
This antibody clone is manufactured by Abcam. If you require a custom buffer formulation or conjugation for your experiments, please contact orders@abcam.com.
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This supplementary information is collated from multiple sources and compiled automatically.
L1CAM also known as neural cell adhesion molecule L1 is a transmembrane protein with a molecular weight of approximately 200-220 kDa. The L1CAM protein belongs to the immunoglobulin superfamily and is heavily glycosylated. It plays a significant role in cell adhesion processes within the nervous system. L1CAM is expressed in a variety of tissues including the brain kidney and peripheral nerves and is notably present in neuronal cells where it engages in cell-cell interactions. Researchers frequently use L1CAM IHC ELISA methods and specific fluorescent markers like Alexa Fluor 568 to study its expression patterns and localization.
L1CAM facilitates neuronal migration axonal growth and synaptic plasticity. This protein participates in the formation and maintenance of neural networks. It interacts with other cell adhesion molecules and components of the extracellular matrix. L1CAM acts as an important modulator within neuronal signaling complexes which impacts the development and regeneration of the nervous system. Insights into its structural and cell interaction properties have made it a focus for understanding nervous system functions.
L1CAM participates extensively in the MAPK and PI3K/AKT pathways. These pathways contribute to cellular growth survival and programmed cell death. The protein interacts with other molecules such as integrins and FGFRs to propagate signaling cascades important for cellular response mechanisms. These connections help modulate cytoskeletal dynamics influencing cellular adhesion and motility needed during brain development and response to injury.
L1CAM has been associated with hydrocephalus and some cancers. Mutations in the L1CAM gene can lead to L1 syndrome which includes a spectrum of neurological disorders particularly hydrocephalus. In cancer overexpression or altered regulation of L1CAM may correlate with tumor aggressiveness and metastatic potential. L1CAM's association with integrin and cadherin proteins underlines its relevance in pathological states and highlights a potential target for therapeutic interventions.
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This species and application combination has not been tested, but we predict it will work based on strong homology. However, this combination is not covered by our product promise.
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Terms & Conditions.
ab24345 staining L1CAM in primary hippocampal rat neurons/glia, (obtained from Neuromics, cat. no. PC35101), DIV14. cells. The cells were fixed with 4% paraformaldehyde (10 min), permeabilized with 0.1% PBS-Tween for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h. The cells were then incubated overnight at 4°C with ab24345 at 5µg/ml and Anti-beta Tubulin antibody - Loading Control ab6046, Rabbit polyclonal to beta Tubulin - Loading Control. Cells were then incubated with Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) preadsorbed ab150117, Goat polyclonal Secondary Antibody to Mouse IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 dilution (shown in green) and Goat Anti-Rabbit IgG H&L (Alexa Fluor® 594) ab150080, Goat polyclonal Secondary Antibody to Rabbit IgG - H&L (Alexa Fluor® 594) at 1/1000 dilution (shown in pseudocolour red). Nuclear DNA was labelled with DAPI (shown in blue).
Image was acquired with a high-content analyser (Operetta CLS, Perkin Elmer) and a maximum intensity projection of confocal sections is shown.
ab24345 staining L1CAM in primary mouse neurons/glia, DIV14 (prepared from E18 mouse hippocampal brain area, obtained from Transnetyx Tissue by BrainBits, LLC, cat.no. C57EHP) cells. The cells were fixed with 100% methanol (5 min), permeabilized with 0.1% PBS-Tween for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1%PBS-Tween for 1h. The cells were then incubated overnight at 4°C with ab24345 at 1µg/ml and Anti-GFAP antibody ab4674, Chicken polyclonal to GFAP. Cells were then incubated with Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) preadsorbed ab150117, Goat polyclonal Secondary Antibody to Mouse IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 dilution (shown in green) and Goat Anti-Chicken IgY H&L (Alexa Fluor® 594) preadsorbed ab150176, Goat polyclonal Secondary Antibody to Chicken IgY - H&L (Alexa Fluor® 594), pre-adsorbed at 1/1000 dilution (shown in pseudocolour red). Nuclear DNA was labelled with DAPI (shown in blue).
Also suitable in cells fixed with 4% paraformaldehyde (10 min).
Image was acquired with a high-content analyser (Operetta CLS, Perkin Elmer) and a maximum intensity projection of confocal sections is shown.
ab24345 staining L1CAM in undifferentiated Neuro-2A cells (top panel) and TRA-differentiated Neuro-2A cells (bottom panel). The cells were fixed with 100% methanol (5min), permeabilized with 0.1%PBS-Tween for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1%PBS-Tween for 1h. The cells were then incubated overnight at +4°C with ab24345 at 5μg/ml and Anti-beta Tubulin antibody - Loading Control ab6046, Rabbit polyclonal to beta Tubulin - Loading Control, at 1/1000 dilution. Cells were then incubated with Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) preadsorbed ab150117, Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) at 1/1000 dilution (shown in green) and Goat Anti-Rabbit IgG H&L (Alexa Fluor® 594) preadsorbed ab150084, Goat polyclonal Secondary Antibody to Rabbit IgG - H&L (Alexa Fluor® 594) at 1/1000 dilution (shown in pseudocolor red). Nuclear DNA was labelled with DAPI (shown in blue).
Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).
This image was generated using the ascites version of the product.
ab24345 staining L1CAM in undifferentiated PC12 cells (top panel) and NGF-differentiated PC12 cells (bottom panel). The cells were fixed with 100% methanol (5min), permeabilized with 0.1%PBS-Tween for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1%PBS-Tween for 1h. The cells were then incubated overnight at +4°C with ab24345 at 5μg/ml and Anti-beta Tubulin antibody - Loading Control ab6046, Rabbit polyclonal to beta Tubulin - Loading Control, at 1/1000 dilution. Cells were then incubated with Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) preadsorbed ab150117, Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) at 1/1000 dilution (shown in green) and Goat Anti-Rabbit IgG H&L (Alexa Fluor® 594) preadsorbed ab150084, Goat polyclonal Secondary Antibody to Rabbit IgG - H&L (Alexa Fluor® 594) at 1/1000 dilution (shown in pseudocolor red). Nuclear DNA was labelled with DAPI (shown in blue).
Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).
This image was generated using the ascites version of the product.
Immunofluorescence analysis of COS7 cells transfected with full-length L1CAM (left) or truncated L1CAM (right), staining L1CAM (green) with ab24345.
Cells were incubated with primary antibody (1/1000 in 1% goat serum + 0.3% Triton X-100 in PBS) and incubated overnight at 4°C. An AlexaFluor®488-conjugated anti-mouse IgG (1/700) was used as the secondary antibody. Nuclei were counterstained with bisbenzimide (blue).
This image was generated using the ascites version of the product.
IHC image of L1CAM staining in a section of formalin-fixed paraffin-embedded normal rat kidney performed on a Leica BONDTM system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20mins. The section was then incubated with ab24345, 0.5ug/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. The inset secondary-only control image is taken from an identical assay without primary antibody.
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.
This image was generated using the ascites version of the product.
This blot was produced using a 4-12% Bis-tris gel under the MOPS buffer system. The gel was run at 200V for 55 minutes before being transferred onto a Nitrocellulose membrane at 30V for 70 minutes. The membrane was then blocked for an hour using 3% milk before ab24345 and Anti-GAPDH antibody [EPR16891] - Loading Control ab181602 (Rabbit anti-GAPDH loading control) were incubated overnight at 4°C at a 1ug/ml concentration and 1/10000 dilution respectively. Antibody binding was detected using Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773) at 1/20000 dilution for 1 hour at room temperature before imaging.
This image was generated using the ascites version of the product.
All lanes: Western blot - Anti-L1CAM antibody [2C2] (ab24345)
Lane 1: Mouse whole brain tissue lysate at 20 µg
Lane 2: Rat whole brain tissue lysate at 20 µg
Lane 3: Human whole brain tissue lysate at 20 µg
Performed under reducing conditions.
Predicted band size: 140 kDa
Observed band size: 200 kDa, 60 kDa
IHC image of L1CAM staining in a section of formalin-fixed paraffin-embedded normal human kidney* performed on a Leica BONDTM system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20mins. The section was then incubated with ab24345, 0.1ug/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. The inset secondary-only control image is taken from an identical assay without primary antibody.
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.
*Tissue obtained from the Human Research Tissue Bank, supported by the NIHR Cambridge Biomedical Research Centre
This image was generated using the ascites version of the product.
ab24345 recognizes one or two polypeptides of L1 or Ng-CAM corresponding to the full length protein (~200kDa) as well as 60-80 kDa C-terminal cleavage products (as shown in the figure).
This image was generated using the ascites version of the product.
All lanes: Western blot - Anti-L1CAM antibody [2C2] (ab24345) at 1/1000 dilution
All lanes: 30ug CNS protein
Performed under reducing conditions.
Predicted band size: 140 kDa
Observed band size: 200 kDa, 60-80 kDa
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