Overview

  • Product nameAnti-L1CAM antibody [2C2]
    See all L1CAM primary antibodies
  • Description
    Mouse monoclonal [2C2] to L1CAM
  • Specificityab24345 L1[2C2] antibody recognizes one or two polypeptides of L1 or Ng-CAM corresponding to the full length protein (~200 kD) and the 60-80 kD C-terminal cleavage products (as shown). ab24345 will recognize protein after denaturation in the presence of reducing agents and can detect protein in lysates (10-100 micrograms of protein) of the nervous system (such as rat cerebellum - see figure).
  • Tested applicationsSuitable for: ICC/IF, IHC-FoFr, Flow Cyt, WBmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Human
  • Immunogen

    chicken NgCAM protein (ab24345 detects the C-terminus portion of the protein that is conserved with mammalian L1)

  • Positive control
    • rat cerebellum, nervous system tissue
  • General notesL1CAM 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.

Properties

Applications

Our Abpromise guarantee covers the use of ab24345 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes
ICC/IF 1/300 - 1/1000.
IHC-FoFr 1/300 - 1/1000.
Flow Cyt 1/20. (paraformaldehyde or methanol fixed cells)



ab170190-Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody.
WB 1/1000. Detects a band of approximately 200 kDa. Cleavage products observed at 60-80 kDa.

Target

  • FunctionCell adhesion molecule with an important role in the development of the nervous system. Involved in neuron-neuron adhesion, neurite fasciculation, outgrowth of neurites, etc. Binds to axonin on neurons.
  • Involvement in diseaseDefects in L1CAM are the cause of hydrocephalus due to stenosis of the aqueduct of Sylvius (HSAS) [MIM:307000]. Hydrocephalus is a condition in which abnormal accumulation of cerebrospinal fluid in the brain causes increased intracranial pressure inside the skull. This is usually due to blockage of cerebrospinal fluid outflow in the brain ventricles or in the subarachnoid space at the base of the brain. In children is typically characterized by enlargement of the head, prominence of the forehead, brain atrophy, mental deterioration, and convulsions. In adults the syndrome includes incontinence, imbalance, and dementia. HSAS is characterized by mental retardation and enlarged brain ventricles.
    Defects in L1CAM are the cause of mental retardation-aphasia-shuffling gait-adducted thumbs syndrome (MASA) [MIM:303350]; also known as corpus callosum hypoplasia, psychomotor retardation, adducted thumbs, spastic paraparesis, and hydrocephalus or CRASH syndrome. MASA is an X-linked recessive syndrome with a highly variable clinical spectrum. Main clinical features include spasticity and hyperreflexia of lower limbs, shuffling gait, mental retardation, aphasia and adducted thumbs. The features of spasticity have been referred to as complicated spastic paraplegia type 1 (SPG1). Some patients manifest corpus callosum hypoplasia and hydrocephalus. Inter- and intrafamilial variability is very wide, such that patients with hydrocephalus, MASA, SPG1, and agenesis of corpus callosum can be present within the same family.
    Defects in L1CAM are the cause of spastic paraplegia X-linked type 1 (SPG1) [MIM:303350]. Spastic paraplegia is a degenerative spinal cord disorder characterized by a slow, gradual, progressive weakness and spasticity of the lower limbs.
    Note=Defects in L1CAM may contribute to Hirschsprung disease by modifying the effects of Hirschsprung disease-associated genes to cause intestinal aganglionosis.
    Defects in L1CAM are a cause of partial agenesis of the corpus callosum (ACCPX) [MIM:304100]. A syndrome characterized by partial corpus callosum agenesis, hypoplasia of inferior vermis and cerebellum, mental retardation, seizures and spasticity. Other features include microcephaly, unusual facies, and Hirschsprung disease in some patients.
  • Sequence similaritiesBelongs to the immunoglobulin superfamily. L1/neurofascin/NgCAM family.
    Contains 5 fibronectin type-III domains.
    Contains 6 Ig-like C2-type (immunoglobulin-like) domains.
  • Cellular localizationCell membrane.
  • Information by UniProt
  • Database links
  • Alternative names
    • Antigen identified by monoclonal antibody R1 antibody
    • CAML1 antibody
    • CD171 antibody
    • CD171 antigen antibody
    • HSAS antibody
    • HSAS1 antibody
    • Hyd antibody
    • L1 antibody
    • L1 cell adhesion molecule antibody
    • L1-NCAM antibody
    • L1cam antibody
    • L1CAM_HUMAN antibody
    • MASA antibody
    • MIC5 antibody
    • N CAML1 antibody
    • N-CAM-L1 antibody
    • NCAM-L1 antibody
    • NCAML1 antibody
    • Nerve-growth factor-inducible large external glycoprotein antibody
    • Neural cell adhesion molecule L1 antibody
    • NILE antibody
    • OTTHUMP00000025992 antibody
    • S10 antibody
    • SPG1 antibody
    see all

Anti-L1CAM antibody [2C2] images

  • ab24345 (1/500) immunostaining L1CAM  in 4% PFA-fixed rat cerebellum tissue. ML = molecular layer and FT = fiber tract of the cerebellum.

  • Anti-L1CAM antibody [2C2] (ab24345) at 1/1000 dilution + 30ug CNS protein

    Performed under reducing conditions.

    Observed band size : 200 kDa (why is the actual band size different from the predicted?)
    Additional bands at : 60-80 kDa (possible cleavage fragment).

    This image is courtesy of Martin Grumet, Rutgers University, United States

    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).

  • Overlay histogram showing PC12 cells stained with ab24345 (red line). The cells were fixed with 4% paraformaldehyde (10 min) and incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab24345, 1/20 dilution) for 30 min at 22°C. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22°C. Isotype control antibody (black line) was mouse IgG1 [ICIGG1] (ab91353, 2µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed. This antibody gave a positive signal in PC12 cells fixed with methanol (5 min) used under the same conditions.

    Please note that Abcam does not have data for use of this antibody on non-fixed cells. We welcome any customer feedback.
  • 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).

References for Anti-L1CAM antibody [2C2] (ab24345)

This product has been referenced in:
  • Ghazvini M  et al. Cancer stemness in Apc- vs. Apc/KRAS-driven intestinal tumorigenesis. PLoS One 8:e73872 (2013). Read more (PubMed: 24069241) »
  • Donier E  et al. L1CAM binds ErbB receptors through Ig-like domains coupling cell adhesion and neuregulin signalling. PLoS One 7:e40674 (2012). ICC/IF . Read more (PubMed: 22815787) »

See all 13 Publications for this product

Product Wall

Thank you for your enquiry.

I can confirm that ab24345 antibody is sold as ascites. Unpurified antibodies, such as those sold as whole antiserum, ascites or tissue culture supernatant will not have a concentration stated on the datasheet. Anti...

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Abcam guarantees this product to work in the species/application used in this Abreview.
Application Western blot
Sample Mouse Cell lysate - whole cell (Brain - Cerebellar granule cells - DIV3)
Loading amount 30 µg
Specification Brain - Cerebellar granule cells - DIV3
Treatment 100 ng/ml human BDNF for 6 minutes
Gel Running Conditions Reduced Denaturing (8% SDS PAGE)
Blocking step Milk as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: RT°C
Username

Abcam user community

Verified customer

Submitted Mar 12 2013

Thank you for your interest in this antibody. Two customers sent their mouse western blot results, both showing detection of bands that they assumed represented the full-length protein and a fragment. However, we do not guarantee that the antibody will...

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Thank you for contacting us. I am sorry to hear your experiencing problems with our antibody. Could you please let me know what type of sections you are using? As of now, fixed-frozen or paraffin-embedded sections have not been tested with this antibo...

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Abcam guarantees this product to work in the species/application used in this Abreview.
Application Immunocytochemistry/ Immunofluorescence
Sample Mouse Cell (EpiSC)
Specification EpiSC
Fixative Paraformaldehyde
Permeabilization Yes - triton 0.1%
Blocking step Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 4% · Temperature: 4°C
Username

Abcam user community

Verified customer

Submitted Nov 09 2011

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Western blot
Sample Rat Cell lysate - whole cell (Cortical neurons)
Loading amount 30 µg
Specification Cortical neurons
Gel Running Conditions Reduced Denaturing
Blocking step Commercially available protein free blocking reagent as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 100% · Temperature: 24°C
Username

Abcam user community

Verified customer

Submitted Feb 15 2008

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Immunocytochemistry/ Immunofluorescence
Sample Rat Cell (Cortical neurons)
Specification Cortical neurons
Fixative Paraformaldehyde
Permeabilization Yes - 0.1% triton X
Blocking step BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: RT°C
Username

Abcam user community

Verified customer

Submitted Feb 15 2008

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Western blot
Sample Mouse Tissue lysate - whole (Mouse Brain)
Specification Mouse Brain
Gel Running Conditions Reduced Denaturing
Blocking step Milk as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Username

Dr. Li Tian

Verified customer

Submitted Sep 21 2007

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Western blot
Sample Mouse Tissue lysate - whole (superior colliculus)
Loading amount 50 µg
Specification superior colliculus
Gel Running Conditions Reduced Denaturing
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 20°C
Username

Abcam user community

Verified customer

Submitted Aug 01 2007

Thank you for your enquiry. The originator of the antibody stated that you could see the cleavage products and no full-length protein in certain sample types, and recommended running a positive control. The positive controls recommended are rat c...

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Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"