Human NF-L Antibody

Detection of Human and rat NF‑L by Western Blot. Western blot shows lysates of SH‑SY5Y human neuroblastoma cell line, A172 human glioblastoma cell line and rat hyphothalamus. PVDF membrane was probed with 2 µg/mL of Mouse Anti-Human NF‑L Monoclonal Antibody (Catalog # MAB22164) followed by HRP-conjugated Anti-Mouse IgG Secondary Antibody (HAF018). A specific band was detected for NF‑L at approximately 68 kDa (as indicated). This experiment was conducted under reducing conditions and using Western Blot Buffer Group 1.

NF‑L in Human Brain (Hippocampus). NF‑L was detected in immersion fixed paraffin-embedded sections of human brain (hippocampus) using Mouse Anti-Human NF‑L Monoclonal Antibody (Catalog # MAB22164) at 5 µg/mL for 1 hour at room temperature followed by incubation with the Anti-Mouse IgG VisUCyte™ HRP Polymer Antibody (VC001). 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 cytoplasm in neurons. Staining was performed using our protocol for IHC Staining with VisUCyte HRP Polymer Detection Reagents.

NF-L in U251 U251 human myeloma cell line (positive) and K562 human chronic myelogenous leukemia cell line (negative). NF‑L was detected in immersion fixed U251 human myeloma cell line (positive) and K562 human chronic myelogenous leukemia cell line (negative) for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Mouse IgG Secondary Antibody (red; Catalog # NL007) and counterstained with DAPI (blue). Specific staining was localized to cytoplasm. View our protocol for Fluorescent ICC Staining of Non-adherent Cells.

Detection of NF-L in Human Cerebellum by Dual RNAscope® ISH-IHC NEFL mRNA (red) and protein (green/blue) was detected in formalin-fixed paraffin-embedded tissue sections of Human Cerebellum. ACD’s Integrated Co-Detection Workflow was performed using ACD RNAScope Probe Hs-NEFL (Catalog # 468671) and Mouse Anti-Human NEFL Monoclonal Antibody (R&D Systems, Catalog # MAB22164) at 7ug/mL. Tissue was stained using RNAscope® 2.5 HD Detection Reagents-RED (Catalog # 322360), Mouse IgG VisUCyte HRP Polymer Antibody (VC001) and RNAscope® 2.5 LS Green Accessory Pack (Catalog #322550). Tissue was counterstained with 50% hematoxylin (purple).