Human L1CAM Antibody

R&D Systems, part of Bio-Techne | Catalog # MAB7773

Recombinant Monoclonal Antibody.
R&D Systems, part of Bio-Techne

Key Product Details

Species Reactivity

Human

Applications

Flow Cytometry, Immunocytochemistry, Simple Western, Western Blot

Label

Unconjugated

Antibody Source

Recombinant Monoclonal Rabbit IgG Clone # 2702C

View all L1CAM Antibodies »

Product Specifications

Immunogen

Mouse myeloma cell line NS0­ derived Human L1CAM
(Ile20­Glu1120) & (Arg864­Glu1120)
Accession # CAA42508

Specificity

Detects human L1CAM in direct ELISAs.

Clonality

Monoclonal

Host

Rabbit

Isotype

IgG

Scientific Data Images for Human L1CAM Antibody

Detection of Human L1CAM by Western Blot.

Western blot shows lysates of HeLa human cervical epithelial carcinoma cell line and human brain (motor cortex). PVDF membrane was probed with 2 µg/mL of Rabbit Anti-Human L1CAM Monoclonal Antibody (Catalog # MAB7773) followed by HRP-conjugated Anti-Rabbit IgG Secondary Antibody (HAF008). A specific band was detected for L1CAM at approximately 250 kDa (as indicated). This experiment was conducted under reducing conditions and using Western Blot Buffer Group 1.

Detection of L1CAM in HeLa Human Cell Line by Flow Cytometry.

HeLa human cervical epithelial cell line was stained with Rabbit Anti-Human L1CAM Monoclonal Antibody (Catalog # MAB7773, filled histogram) or Rabbit IgG isotype control antibody (MAB1050, open histogram), followed by Phycoerythrin-conjugated Anti-Rabbit IgG Secondary Antibody (F0110). Staining was performed using our Staining Membrane-associated Proteins protocol.

L1CAM in HeLa Human Cell Line.

L1CAM was detected in immersion fixed HeLa human cervical epithelial carcinoma cell line (positive staining) and Daudi human Burkitt's lymphoma cell line (negative staining) using Rabbit Anti-Human L1CAM Monoclonal Antibody (Catalog # MAB7773) at 3 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Rabbit IgG Secondary Antibody (red; NL004) and counterstained with DAPI (blue). Specific staining was localized to cell membrane. Staining was performed using our protocol for Fluorescent ICC Staining of Non-adherent Cells.
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Applications for Human L1CAM Antibody

Application
Recommended Usage

Flow Cytometry

0.25 µg/106 cells
Sample: HeLa human cervical epithelial cell line

Immunocytochemistry

3-25 µg/mL
Sample: Immersion fixed HeLa human cervical epithelial carcinoma cell line and MCF‑7 human breast cancer cell line

Simple Western

20 µg/mL
Sample: HeLa human cervical epithelial carcinoma cell line

Western Blot

2 µg/mL
Sample: HeLa human cervical epithelial carcinoma cell line and human brain (motor cortex)

Formulation, Preparation, and Storage

Purification

Protein A or G purified from cell culture supernatant

Reconstitution

Reconstitute at 0.5 mg/mL in sterile PBS. For liquid material, refer to CoA for concentration.

Reconstitution Buffer Available:
Size / Price
Qty

Formulation

Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied either lyophilized or as a 0.2 µm filtered solution in PBS.

Shipping

Lyophilized product is shipped at ambient temperature. Liquid small pack size (-SP) is shipped with polar packs. Upon receipt, store immediately at the temperature recommended below.

Stability & Storage

Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
  • 12 months from date of receipt, -20 to -70 °C as supplied.
  • 1 month, 2 to 8 °C under sterile conditions after reconstitution.
  • 6 months, -20 to -70 °C under sterile conditions after reconstitution.

Background: L1CAM

L1CAM (Neural cell adhesion molecule L1, also known as L1, CD171 and NCAM-L1) is a founding member of the L1 family, Immunoglobulin (Ig) superfamily of molecules (1-4). It was initially described as a 200-230 kDa neural adhesion molecule that likely played a key role in mouse nervous system development (4-6). Subsequent studies have confirmed the adhesive nature of the molecule, and expanded its activities in both neural and nonneural cell types. L1 is now recognized to play a key role in cell migration, adhesion, neurite outgrowth, myelination and neuronal differentiation (1, 7, 8). It does so through a series of cis and trans interactions that involve multiple copartners and target receptors (1, 3, 6, 8). Cells known to express L1 are varied, and include immature oligodendrocytes (9), CD4+ T cells, B cells and monocytes (10), both motor and sensory Schwann cells (11, 12), intestinal epithelial progenitor cells (12), cerebellar granule and Purkinje cells (5, 13, 14), and multiple tumor cells such as melanoma (15) plus pancreatic duct and lung carcinoma cells (16, 17). Human L1 was first identified as a 215 kDa glycoprotein on the surface of SKNAS neuroblastoma cells (18). Subsequent cloning established its precursor as being 1257 amino acids (aa) in length (19, 20). The molecule is a type I transmembrane (TM) protein that contains an 1101 aa extracellular region (aa 20-1120) plus a 114 aa cytoplasmic domain (aa 1144-1257). The extracellular region possesses six C2-type Ig-like domains (aa 35-607) followed by five fibronectin (FN) type III domains (aa 612-1108). As noted, L1 participates in multiple cis and trans interactions, and some of these interaction sites have been mapped to select Ig or FN domains. For instance, Ig-like domains #1, 2 and 6 associate with NP-1, L1 (homotypic binding), and various integrins, respectively (21‑23). The latter interaction is mediated by one (in human) or two (in mouse) RGD motifs (23, 24). Other molecules that heterotypically associate with L1 include NCAM, neurocan, CD24 and EGFR. The cytoplasmic tail contains no kinase motifs, but does possess a FIGQY peptide that interacts with ankyrin, and an RSLE sequence that mediates clathrin-associated endocytosis (1). There are two splice variants, one each in the intracellular and extracellular domains.  A deletion of RSLE adversely affects endocytosis, while a Leu substitution for aa 26-31 interfers with numerous heterotypic interactions (25, 26).  In general, the full-length L1 molecule is a neuron-associated isoform. L1 is known to undergo proteolysis, either by plasmin or ADAMs. This generates soluble isoforms of varying sizes (140-200 kDa) that retain bioactivity, and which can be incorporated into the surrounding ECM (5, 13, 27-30). The membrane fragments (30-80 kDa) undergo further processing, most importantly by gamma-secretase, to generate a soluble 28 kDa intracellular domain. This domain is SUMOylated, and believed to possess an NLS at Lys1147.  Upon presumed entry into the nucleus, L1 is posited to activate L1-responsive genes. Human and mouse L1 precursors share 88% aa sequence identity.

References

  1. Maness, P.F. and M. Schachner (2007) Nat. Neurosci. 10:19.
  2. Wei, C.H. and S.E. Ryu (2012) Exp. Mol. Med. 44:413.
  3. Faspel, J. and M. Grumet (2003) Front. Biosci. 8:1210.
  4. Herron, L.R. et al. (2009) Biochem. J. 419:519.
  5. Rathjen, F.G. and M. Schachner (1984) EMBO. J. 3:1.
  6. Keifel, H. et al. (2011) Trends Mol. Med. 17:178.
  7. Dihne, M. et al. (2003) J. Neurosci. 23:6638.
  8. Kadmon, G. et al. (1998) Dev. Immunol. 6:205.
  9. Itoh, K. et al. (2000) J. Neurosci. Res. 60:579.
  10. Jouet, M. et al. (1995) Mol. Brain Res. 30:378.
  11. He, Q. et al. (2012) Neurosci. Lett. 521:57.
  12. Thor, G. et al. (1987) EMBO J. 6:2581.
  13. Sadoul, K. et al. (1988) J. Neurochem. 50:510.
  14. Hubbe, M. et al. (1993) Eur. J. Immunol. 23:2927.
  15. Hoja-Lukowicz, D. et al. (2012) Glycoconj J. Apr 29. [Epub ahead of print].
  16. Geismann, C. et al. (2009) Cancer Res. 69:4517.
  17. Tischler, V. et al. (2011) Mol. Cancer 10:127.
  18. Mujoo, K. et al. (1986) J. Biol. Chem. 261:10299.
  19. Kobayashi, M. et al. (1991) Biochim. Biophys. Acta 1090:238.
  20. Hlavin, M.L. and V. Lemmon (1991) Genomics 11:416.
  21. Castellani, V. et al. (2002) EMBO J. 21:6348.
  22. Zhao, X. and C-H Siu (1995) J. Biol. Chem. 270:29413.
  23. Oleszewski, M. et al. (1999) J. Biol. Chem. 274:24602.
  24. Felding-Habermann, B. et al. (1997) J. Cell Biol. 139:1567.
  25. Kamiguchi, H. et al. (1998) J. Neurosci. 18:5311.
  26. De Angelis, E. et al. (2001) J. Biol. Chem. 276:32738.
  27. Montgomery, A.M.P. et al. (1996) J. Cell Biol. 132:475.
  28. Sadoul, R. et al. (1989) J. Neurochem. 53:1471.
  29. Reidle, S. et al. (2009) Biochem. J. 420:391.
  30. Lutz, D. et al. (2012) J. Biol. Chem. 287:17161.

Long Name

Cell Adhesion Molecule L1

Alternate Names

CAML1, CD171, HSAS, HSAS1, MASA, MIC5, NCAM-L1, S10, SPG1

Entrez Gene IDs

3897 (Human); 16728 (Mouse); 50687 (Rat)

Gene Symbol

L1CAM

UniProt

CAA42508

Additional L1CAM Products

Product Documents for Human L1CAM Antibody

Product Datasheets

Certificate of Analysis

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Product Specific Notices for Human L1CAM Antibody

For research use only

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