Name: Anti-L1CAM antibody [EPR18750]
Brand: RabMAb
SKU: ab208155
Rating: 4 (2 reviews)

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Western blot - Anti-L1CAM antibody [EPR18750] (ab208155)

Key features and details

Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
Rabbit monoclonal [EPR18750] to L1CAM
Suitable for: WB, IHC-P, IP
Knockout validated
Reacts with: Mouse, Rat, Human

Carrier Free

Product name

Anti-L1CAM antibody [EPR18750]
See all L1CAM primary antibodies
Description

Rabbit monoclonal [EPR18750] to L1CAM
Host species

Rabbit
Tested applications

Suitable for: WB, IHC-P, IPmore details
Species reactivity

Reacts with: Mouse, Rat, Human
Immunogen

Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.
Positive control
- WB: Human fetal brain and cerebellum lysates; HeLa and A-375 whole cell lysates; Rat brain, cerebellum and hippocampus lysates. Mouse cerebellum and brain lysates. IHC-P: Human kidney, Human stomach cancer, Mouse cerebrum, Mouse colon, Rat cerebellum and Rat colon tissues. IP: Human cerebellum and Rat brain lysates.
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
For more information see here.
Our RabMAb^® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb^® patents.

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, 0.05% BSA
Concentration information loading...
Purity

Protein A purified
Clonality

Monoclonal
Clone number

EPR18750
Isotype

IgG
Research areas
- Neuroscience
- Development

Alternative Versions
- Anti-L1CAM antibody [EPR18750] - Low endotoxin, Azide free (ab213611)
- Anti-L1CAM antibody [EPR18750] - BSA and Azide free (ab271982)
Compatible Secondaries
- Goat Anti-Rabbit IgG H&L (HRP) (ab97051)
Isotype control
- Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)
KO cell lines
- Human L1CAM knockout HeLa cell line (ab255401)
KO cell lysates
- Human L1CAM knockout HeLa cell lysate (ab263786)
Recombinant Protein
- Recombinant Human L1CAM protein (His tag) (ab222980)
Related Products
- VeriBlot for IP Detection Reagent (HRP) (ab131366)

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab208155 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
WB	(1)	1/1000. Predicted molecular weight: 140 kDa.
IHC-P	(1)	1/500. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
IP		1/40.

Notes
WB 1/1000. Predicted molecular weight: 140 kDa.
IHC-P 1/500. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
IP 1/40.

Function

Cell 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 disease

Defects 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 similarities

Belongs to the immunoglobulin superfamily. L1/neurofascin/NgCAM family.
Contains 5 fibronectin type-III domains.
Contains 6 Ig-like C2-type (immunoglobulin-like) domains.
Cellular localization

Cell membrane.
Target information above from: UniProt accession P32004 The UniProt Consortium
The Universal Protein Resource (UniProt) in 2010
Nucleic Acids Res. 38:D142-D148 (2010) .

Information by UniProt
Database links
- Entrez Gene: 3897 Human
- Entrez Gene: 16728 Mouse
- Entrez Gene: 50687 Rat
- Omim: 308840 Human
- SwissProt: P32004 Human
- SwissProt: P11627 Mouse
- SwissProt: Q05695 Rat
- Unigene: 522818 Human
- Unigene: 260568 Mouse
- Unigene: 10378 Rat
see all
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

Western blot - Anti-L1CAM antibody [EPR18750] (ab208155)

All lanes : Anti-L1CAM antibody [EPR18750] (ab208155) at 1/1000 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 kDa

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. 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. 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.
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.
Western blot - Anti-L1CAM antibody [EPR18750] (ab208155)

All lanes : Anti-L1CAM antibody [EPR18750] (ab208155) at 1/2000 dilution

Lane 1 : Human fetal brain lysate
Lane 2 : Human cerebellum lysate
Lane 3 : HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate

Lysates/proteins at 20 µg per lane.

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: 140,200-220 kDa why is the actual band size different from the predicted?

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.
Western blot - Anti-L1CAM antibody [EPR18750] (ab208155)

All lanes : Anti-L1CAM antibody [EPR18750] (ab208155) at 1/2000 dilution

Lane 1 : Rat brain lysate
Lane 2 : Rat cerebellum lysate
Lane 3 : Rat hippocampus lysate

Lysates/proteins at 20 µg per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

Predicted band size: 140 kDa
Observed band size: 140,200-220 kDa why is the actual band size different from the predicted?

Exposure time: 3 minutes

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.
Western blot - Anti-L1CAM antibody [EPR18750] (ab208155)

All lanes : Anti-L1CAM antibody [EPR18750] (ab208155) at 1/1000 dilution

Lane 1 : Mouse cerebellum lysate
Lane 2 : Mouse brain lysate
Lane 3 : A-375 (Human malignant melanoma cell line) whole cell lysate

Lysates/proteins at 10 µg per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

Predicted band size: 140 kDa
Observed band size: 140,200-220 kDa why is the actual band size different from the predicted?

Exposure time: 3 minutes

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

Western blot protocols
Immunoprecipitation protocols
Immunohistochemistry protocols

Click here to view the general protocols

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Publishing research using ab208155? Please let us know so that we can cite the reference in this datasheet.

ab208155 has been referenced in 5 publications.

Yamamoto-Fukuda T et al. L1CAM-ILK-YAP Mechanotransduction Drives Proliferative Activity of Epithelial Cells in Middle Ear Cholesteatoma. Am J Pathol N/A:N/A (2020). PubMed: 32360569
Yang H et al. Downregulation of lncRNA XIST Represses Tumor Growth and Boosts Radiosensitivity of Neuroblastoma via Modulation of the miR-375/L1CAM Axis. Neurochem Res 45:2679-2690 (2020). PubMed: 32857295
Shikanai M et al. Caveolin-1 Promotes Early Neuronal Maturation via Caveolae-Independent Trafficking of N-Cadherin and L1. iScience 7:53-67 (2018). PubMed: 30267686
Agrawal P et al. A Systems Biology Approach Identifies FUT8 as a Driver of Melanoma Metastasis. Cancer Cell 31:804-819.e7 (2017). PubMed: 28609658
Fu TG et al. miR-143 inhibits oncogenic traits by degrading NUAK2 in glioblastoma. Int J Mol Med 37:1627-35 (2016). WB ; Human . PubMed: 27081712

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Application

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)

Sample

Mouse Tissue sections (MC38 murine colon cancer cell)

Antigen retrieval step

Heat mediated - Buffer/Enzyme Used: Tris/EDTA

Permeabilization

Specification

MC38 murine colon cancer cell

Blocking step

Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C

Fixative

Formaldehyde

The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

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Submitted Dec 24 2019

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Key features and details

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Overview

Product name

Description

Host species

Tested applications

Species reactivity

Immunogen

Positive control

General notes

Properties

Form

Storage instructions

Storage buffer

Purity

Clonality

Clone number

Isotype

Research areas

Associated products

Alternative Versions

Compatible Secondaries

Isotype control

KO cell lines

KO cell lysates

Recombinant Protein

Related Products

Applications

The Abpromise guarantee

Target

Function

Involvement in disease

Sequence similarities

Cellular localization

Database links

Alternative names

Images

Protocols

Datasheets and documents

SDS download

Datasheet download

Certificate of Compliance

References (5)

Customer reviews and Q&As

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