CHL-1/L1CAM-2 Antibody (316223) [Unconjugated]

CHL‑1/L1CAM‑2 was detected in immersion fixed paraffin-embedded sections of human melanoma tissue using Rat Anti-Human CHL‑1/L1CAM‑2 Monoclonal Antibody (Catalog # MAB2126) at 1.7 µg/mL for 1 hour at room temperature followed by incubation with the Anti-Rat IgG VisUCyte™ HRP Polymer Antibody (VC005). Before incubation with the primary antibody, tissue was subjected to heat-induced epitope retrieval using Antigen Retrieval Reagent-Basic (CTS013). Tissue was stained using DAB (brown) and counterstained with hematoxylin (blue). Specific staining was localized to cell nuclei. Staining was performed using our protocol for IHC Staining with VisUCyte HRP Polymer Detection Reagents.

CHL‑1/L1CAM‑2 was detected in immersion fixed paraffin-embedded sections of human spleen using Rat Anti-Human CHL‑1/L1CAM‑2 Monoclonal Antibody (Catalog # MAB2126) at 1.7 µg/mL for 1 hour at room temperature followed by incubation with the Anti-Rat IgG VisUCyte™ HRP Polymer Antibody (VC005). Before incubation with the primary antibody, tissue was subjected to heat-induced epitope retrieval using Antigen Retrieval Reagent-Basic (CTS013). Tissue was stained using DAB (brown) and counterstained with hematoxylin (blue). Specific staining was localized to cell nuclei. Staining was performed using our protocol for IHC Staining with VisUCyte HRP Polymer Detection Reagents.

Western blot analysis of the protein levels of CHL1 detected in normal human glial HEB cells and 3 glioma/glioblastoma cell lines. CHL1 was weakly expressed in normal human HEB glial cells. Its levels in all the 3 glioma/glioblastoma cells were higher than that in normal human HEB glial cells, with the statistical significance detected in SHG44 cells (*p < 0.05 vs. HEB cells) and U-87 MG cells (**p < 0.01 vs. HEB cells). n = 3 for each group. Student’s t-test for independent samples was used. Image collected and cropped by CiteAb from the following publication (//journal.frontiersin.org/article/10.3389/fnmol.2017.00324/full), licensed under a CC-BY license. Not internally tested by R&D Systems.

H&E staining and immunohistochemical staining analyses for the CHL1, caspase-3, PCNA and GFAP molecules in glioblastoma xenograft tissues from both control siRNA and CHL1 siRNA-treated groups. Scale bars represent 25 μm. Image collected and cropped by CiteAb from the following publication (//journal.frontiersin.org/article/10.3389/fnmol.2017.00324/full), licensed under a CC-BY license. Not internally tested by R&D Systems.

Treatment of siRNA targeting CHL1 in three human glioma cell lines. Total RNA was isolated from U251, SHG44 and U-87 MG cells treated with vehicle control (vc), control siRNA (control siRNA) or siRNA targeting CHL1 (CHL1 siRNA). RT-PCR and Western blot analysis were then used to measure both relative mRNA and protein levels of CHL1. (A) RT-PCR analysis of the mRNA levels of CHL1 in U251, SHG44 and U-87 MG cells treated with vehicle control (vc), control siRNA and siRNA targeting CHL1, and (B) Western blot analysis of the protein levels of CHL1 detected in U251, SHG44 and U-87 MG cells treated with vehicle control (vc), control siRNA and siRNA targeting CHL1. Data are presented as means ± standard error of the mean (SEM) (n = 3, *p < 0.05; **p < 0.01, independent Student’s t-test). Image collected and cropped by CiteAb from the following publication (//journal.frontiersin.org/article/10.3389/fnmol.2017.00324/full), licensed under a CC-BY license. Not internally tested by R&D Systems.

Treatment of siRNA targeting CHL1 in three human glioma cell lines. Total RNA was isolated from U251, SHG44 and U-87 MG cells treated with vehicle control (vc), control siRNA (control siRNA) or siRNA targeting CHL1 (CHL1 siRNA). RT-PCR and Western blot analysis were then used to measure both relative mRNA and protein levels of CHL1. (A) RT-PCR analysis of the mRNA levels of CHL1 in U251, SHG44 and U-87 MG cells treated with vehicle control (vc), control siRNA and siRNA targeting CHL1, and (B) Western blot analysis of the protein levels of CHL1 detected in U251, SHG44 and U-87 MG cells treated with vehicle control (vc), control siRNA and siRNA targeting CHL1. Data are presented as means ± standard error of the mean (SEM) (n = 3, *p < 0.05; **p < 0.01, independent Student’s t-test). Image collected and cropped by CiteAb from the following publication (//journal.frontiersin.org/article/10.3389/fnmol.2017.00324/full), licensed under a CC-BY license. Not internally tested by R&D Systems.

• Reactivity: Hu
• Applications: WB, IHC
• Clone: 316223
• Clonality: Monoclonal
• Host: Rat
• Conjugate: Unconjugated
• Concentration: LYOPH

CHL-1/L1CAM-2 Antibody (316223) [Unconjugated] Summary

• Immunogen: Mouse myeloma cell line NS0-derived recombinant human CHL‑1/L1CAM‑2
  Ile25-Gln1096
  Accession # NP_006605
• Specificity: Detects human CHL‑1/L1CAM‑2 in direct ELISAs and Western blots. In direct ELISAs and Western blots, no cross-reactivity with recombinant mouse CHL-1 is observed.
• Source: N/A
• Isotype: IgG1
• Clonality: Monoclonal
• Host: Rat
• Gene: CHL1
• Purity Statement: Protein A or G purified from hybridoma culture supernatant

Applications/Dilutions

• Dilutions:
  • Immunohistochemistry 1.7-25 ug/mL
  • Western Blot 1 ug/mL

Packaging, Storage & Formulations

• 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.
• Buffer: 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.
• Preservative: No Preservative
• Concentration: LYOPH
• Reconstitution Instructions: Reconstitute at 0.5 mg/mL in sterile PBS.

Notes

This product is produced by and ships from R&D Systems, Inc., a Bio-Techne brand.

Alternate Names for CHL-1/L1CAM-2 Antibody (316223) [Unconjugated]

• CALL
• CALLClose homolog of L1
• cell adhesion molecule with homology to L1CAM (close homolog of L1)
• cell adhesion molecule with homology to L1CAM (close homologue of L1)
• CHL1
• CHL-1
• FLJ44930
• L1CAM-2
• L1CAM2L1 cell adhesion molecule 2
• MGC132578
• neural cell adhesion molecule L1-like protein

Background

Close homolog of L1 (CHL-1), also known as cell adhesion L1-like (CALL) and L1 cell adhesion molecule 2 (L1-CAM2), belongs to the L1 subfamily of the Ig superfamily cell adhesion molecules, which also include L1, neurofascin and NgCAM-related cell adhesion molecule (NrCAM) (1‑3). These molecules are type I transmembrane proteins that have 6 Ig-like domains and 4‑5 fibronectin type III-like (FNIII) domains in their extracellular regions. They also shared a highly conserved cytoplasmic region of approximately 110 amino acids (aa) containing an ankyrin-binding site. CHL-1 is expressed as a highly glycosylated 185 kDa transmembrane protein by subpopulations of neurons and glia of the central and peripheral nervous system (4, 5). Ectodomain shedding via the metalloprotease-disintegrin ADAM8 releases 165 kDa and 125 kDa soluble CHL-1 fragments, which can diffuse away to function at distant sites (6). CHL-1 is not capable of homotypic interactions, but an extracellular binding partner of CHL-1 has not been identified (4). Human CHL1 has been mapped to chromosome 3p26 and is a candidate gene for 3p^- syndrome characterized by mental impairment (7). A missense CHL1 polymorphism associated with an increased risk of schizophrenia has been reported (8). The functional importance of CHL-1 in the nervous system is also evident in CHL-1 deficient mice, which display behavioral abnormalities and show misguided axons within the hippocampus and olfactory tract (9). Enhanced ectodomain-shedding of CHL-1 is also observed in Wobbler mice, the neurodegenerative mutant mice (6). In vitro, soluble or substrate-coated CHL-1 promotes neurite outgrowth and neuronal survival of both cerebellar and hippocampal neurons. Cell surface CHL-1 interacts with integrins in cis to potentiate integrin-dependent cell migration toward extracellular matrix proteins (10). For this enhanced cell motility, CHL-1 linkage to the actin cytoskeleton via interaction between ankyrin and the CHL-1 cytoplasmic region is required.

1. Moos, M. et al. (1988) Nature 334:701. 
2. Holm, J. et al. (1996) Eur. J. Neusci. 8:1613. 
3. Wei, M. et al. (1998) Hum. Genet. 103:355. 
4. Hillenbrand, R. et al. (1999) Eur. J. Neurosci. 11:813. 
5. Liu, Q. et al. (2000) J. Neurosci. 20:7682. 
6. Naus, S. et al. (2004) J. Biol. Chem. 279:16083.
7. Angeloni, D. et al. (1999) Am. J. Med. Genet. 86:482.
8. Sakurai, K. et al. (2002) Mol. Psychiatry 7:412.
9. Montag-Sallaz, M. et al. (2002) Mol. Cell. Biol. 22(22):7967.
10. Buhusi, M. et al. (2003) J. Biol. Chem. 278(27):25024.

Limitations

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

Publications for CHL-1/L1CAM-2 Antibody (MAB2126)(7)

Publications using MAB2126

• Simoes S, Neufeld J. L, et al. Tau and other proteins found in Alzheimer's disease spinal fluid are linked to retromer-mediated endosomal traffic in mice and humans. Sci Transl Med 11/25/2020 [PMID: 33239387]
• WW Lin, GY Ou, JZ Lin, SJ Yi, WC Yao, HC Pan, WJ Zhao Neuregulin 1 enhances cell adhesion molecule L1 like expression levels and promotes malignancy in human glioma Oncol Lett, 2020-04-21;20(1):326-336. 2020-04-21 [PMID: 32565959] (ICC, IHC, Western Blot, Human)
• KF Karstens, E Bellon, A Polonski, G Wolters-Ei, N Melling, M Reeh, JR Izbicki, M Tachezy Expression and serum levels of the neural cell adhesion molecule L1-like protein (CHL1) in gastrointestinal stroma tumors (GIST) and its prognostic power Oncotarget, 2020-03-31;11(13):1131-1140. 2020-03-31 [PMID: 32284790] (ELISA Capture, Human)
• Norihiro Kotani, Arisa Yamaguchi, Tomoko Ohnishi, Ryusuke Kuwahara, Takanari Nakano, Yuka Nakano et al. Proximity proteomics identifies cancer cell membrane cis ‐molecular complex as a potential cancer target Cancer Science 8/1/2019 [PMID: 31228215]
• W Kim, L Ma, S Lomoio, R Willen, S Lombardo, J Dong, PG Haydon, G Tesco BACE1 elevation engendered by GGA3 deletion increases ?-amyloid pathology in association with APP elevation and decreased CHL1 processing in 5XFAD mice Mol Neurodegener, 2018-02-02;13(1):6. 2018-02-02 [PMID: 29391027] (Western Blot, Human)
• Zhai Yang, Qing Xie, Cheng-Liang Hu, Qiong Jiang, Hui-Fan Shen, Melitta Schachner et al. CHL1 Is Expressed and Functions as a Malignancy Promoter in Glioma Cells Frontiers in Molecular Neuroscience 10/17/2017 [PMID: 29089868]
• Wolterink S, Moldenhauer G, Fogel M, Kiefel H, Pfeifer M, Luttgau S, Gouveia R, Costa J, Endell J, Moebius U, Altevogt P Therapeutic antibodies to human L1CAM: functional characterization and application in a mouse model for ovarian carcinoma. Cancer Res., 2010-03-09;70(6):2504-15. 2010-03-09 [PMID: 20215505] (Blocking, Human)

Bioinformatics

• Gene Symbol: CHL1
• Uniprot:
  • NP_006605()