Human CHL-1/L1CAM-2 Antibody

CHL‑1/L1CAM‑2 in Human Melanoma Tissue. 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 in Human Spleen. 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.

Detection of Human CHL-1/L1CAM-2 by Western Blot 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 (https://journal.frontiersin.org/article/10.3389/fnmol.2017.00324/full), licensed under a CC-BY license. Not internally tested by R&D Systems.

Detection of Human CHL-1/L1CAM-2 by Immunohistochemistry 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 (https://journal.frontiersin.org/article/10.3389/fnmol.2017.00324/full), licensed under a CC-BY license. Not internally tested by R&D Systems.

Detection of Human CHL-1/L1CAM-2 by Western Blot 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 (https://journal.frontiersin.org/article/10.3389/fnmol.2017.00324/full), licensed under a CC-BY license. Not internally tested by R&D Systems.

Detection of Human CHL-1/L1CAM-2 by Western Blot 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 (https://journal.frontiersin.org/article/10.3389/fnmol.2017.00324/full), licensed under a CC-BY license. Not internally tested by R&D Systems.