L1CAM Antibody (Rabbit mAb) [M4J1]

Application: Reactivity: Human

Usage Information

Dilution
WB1:1000 IP1:30 IHC1:100 IF1:100 - 1:500 FCM1:50 - 1:500

Application
WB, IP, IHC, IF, FCM

Reactivity
Human

Source
Rabbit Monoclonal Antibody

Storage Buffer
PBS, pH 7.2+50% Glycerol+0.05% BSA+0.01% NaN3

Storage (from the date of receipt)
-20°C (avoid freeze-thaw cycles), 2 years

Predicted MW Observed MW
140 kDa 250 kDa, 80 kDa
^*Why do the predicted and actual molecular weights differ? The following reasons may explain differences between the predicted and actual protein molecular weight. Post-translational modifications(e.g., phosphorylation, glycosylation); Splice variants and isoforms; Relative charge; Multimerization.

Positive Control	Human spleen tissue; Human cerebellum tissue; Human kidney tissue; Human brain tissue; HeLa cells; Parental HeLa; A375 cells
Negative Control	MCF7 cells

Datasheet & SDS

Biological Description

Specificity
L1CAM Antibody (Rabbit mAb) [M4J1] detects endogenous levels of total L1CAM protein.

Clone
M4J1

Synonym(s)
CD171, CAML1, MIC5, L1CAM, Neural cell adhesion molecule L1, N-CAM-L1, NCAM-L1

Background
L1CAM belongs to the immunoglobulin superfamily of cell adhesion molecules and is built as a type I transmembrane glycoprotein composed of six immunoglobulin-like domains followed by five fibronectin type III repeats, a single transmembrane region, and a highly conserved cytoplasmic tail; this modular ectodomain supports two distinct binding modes, homophilic engagement with L1CAM molecules on adjacent cells and heterophilic binding to other neural cell adhesion molecules, integrins, CD24, neurocan, and neuropilin-1. Within the sixth immunoglobulin domain, an RGD motif mediates direct binding to integrins including alpha5beta1, alphavbeta3, and alphavbeta5, and this integrin engagement activates signaling distinct from that triggered by homophilic L1CAM-L1CAM binding. Homophilic binding functions predominantly in a static, adhesive capacity, gluing adjacent cells together and activating the MAPK pathway, and this MAPK signal can be further modulated through coupling to growth factor receptors known to interact with L1CAM. In contrast, L1CAM-integrin engagement drives a motility-promoting function by activating NF-kB signaling, which supports cell migration and invasiveness, so that the same protein produces functionally opposite cellular behaviors, static adhesion versus active migration, depending on which binding partner it engages. A key determinant of this functional switch is proteolytic processing: L1CAM is cleaved proximal to the plasma membrane by the metalloproteinases ADAM10 and ADAM17, releasing a soluble ectodomain fragment while leaving a smaller transmembrane stub in the membrane, and this ectodomain shedding, together with the capacity to exchange binding partners between homophilic and integrin-mediated modes, governs the transition from the adhesive to the migratory functional state. The cytoplasmic tail of L1CAM engages the cytoskeletal proteins ankyrin, spectrin, actin, and the ERM protein ezrin, and this ezrin linkage is the principal route through which full-length, membrane-associated L1CAM drives ERK activation, proliferation, and gene regulation, while the cleaved cytoplasmic fragment can additionally translocate to the nucleus and promote gene transcription through an ERK-independent mechanism, extending L1CAM's signaling output beyond the plasma membrane. Loss-of-function L1CAM mutations produce L1 syndrome, a spectrum of neurodevelopmental disorders including hydrocephalus, and elevated L1CAM expression correlates with advanced tumor stage, metastasis, and poor prognosis across multiple cancers.

Background

L1CAM belongs to the immunoglobulin superfamily of cell adhesion molecules and is built as a type I transmembrane glycoprotein composed of six immunoglobulin-like domains followed by five fibronectin type III repeats, a single transmembrane region, and a highly conserved cytoplasmic tail; this modular ectodomain supports two distinct binding modes, homophilic engagement with L1CAM molecules on adjacent cells and heterophilic binding to other neural cell adhesion molecules, integrins, CD24, neurocan, and neuropilin-1. Within the sixth immunoglobulin domain, an RGD motif mediates direct binding to integrins including alpha5beta1, alphavbeta3, and alphavbeta5, and this integrin engagement activates signaling distinct from that triggered by homophilic L1CAM-L1CAM binding. Homophilic binding functions predominantly in a static, adhesive capacity, gluing adjacent cells together and activating the MAPK pathway, and this MAPK signal can be further modulated through coupling to growth factor receptors known to interact with L1CAM. In contrast, L1CAM-integrin engagement drives a motility-promoting function by activating NF-kB signaling, which supports cell migration and invasiveness, so that the same protein produces functionally opposite cellular behaviors, static adhesion versus active migration, depending on which binding partner it engages. A key determinant of this functional switch is proteolytic processing: L1CAM is cleaved proximal to the plasma membrane by the metalloproteinases ADAM10 and ADAM17, releasing a soluble ectodomain fragment while leaving a smaller transmembrane stub in the membrane, and this ectodomain shedding, together with the capacity to exchange binding partners between homophilic and integrin-mediated modes, governs the transition from the adhesive to the migratory functional state. The cytoplasmic tail of L1CAM engages the cytoskeletal proteins ankyrin, spectrin, actin, and the ERM protein ezrin, and this ezrin linkage is the principal route through which full-length, membrane-associated L1CAM drives ERK activation, proliferation, and gene regulation, while the cleaved cytoplasmic fragment can additionally translocate to the nucleus and promote gene transcription through an ERK-independent mechanism, extending L1CAM's signaling output beyond the plasma membrane. Loss-of-function L1CAM mutations produce L1 syndrome, a spectrum of neurodevelopmental disorders including hydrocephalus, and elevated L1CAM expression correlates with advanced tumor stage, metastasis, and poor prognosis across multiple cancers.

References
https://pubmed.ncbi.nlm.nih.gov/22796939/ https://pubmed.ncbi.nlm.nih.gov/31455004/

Protein Size (kDa)	PVDF Transfer Membrane Pore Size (μm)
< 10	0.22
10-20	0.22
20-30	0.45
30-45	0.45
45-70	0.45
80-100	0.45
100-140	0.45
> 140	0.45

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Protein Size (kDa)	Separating Gel Percentage
4-40	20%
12-45	15%
10-70	12.5%
15-100	10%
25-100	8%
60-210	5%