Western blotting analysis using neurofilament medium chain antibody (Cat#63480). Total cell lysates (30 μg) from various cell lines were loaded and separated by SDS-PAGE. The blot was incubated with neurofilament medium chain antibody (Cat#63480, 1:5,000) and HRP-conjugated goat anti-mouse secondary antibody (Cat#101, 1:20,000) respectively. Image was developed using FeQ™ ECL Substrate Kit (Cat#226).
Western blotting analysis using neurofilament medium chain antibody (Cat#63480). Neurofilament medium chain expression in wild type (WT) and neurofilament medium chain (NEFM) shRNA knockdown (KD) HeLa cells with 20 μg of total cell lysates. β-Tubulin serves as a loading control. The blot was incubated with neurofilament medium chain antibody (Cat#63480, 1:5,000) and HRP-conjugated goat anti-mouse secondary antibody (Cat#101, 1:20,000) respectively. Image was developed using NaQ™ ECL Substrate Kit (Cat#716).
Immunohistochemistry was performed on paraffin-embedded human skin tissue using neurofilament medium chain antibody (Cat#63480, 1:200). Antigen retrieval was done in sodium citrate buffer (pH 6.0). DAB was used for detection, with hematoxylin counterstaining. Images were acquired using a Nikon Ci-L Plus microscope (40× objective). Scale bar, 25 μm.
Immunocytochemical staining of HeLa cells with neurofilament medium chain antibody (Cat#63480, 1:1,000) . Nuclei were stained blue with DAPI; Neurofilament medium chain was stained magenta with Alexa Fluor® 647. Images were taken using Leica stellaris 5. Protein abundance based on laser Intensity and Smart Gain: High. Scale bar, 20 μm.
Immunocytochemical staining of HeLa cells using neurofilament medium chain antibody (Cat#63480, 1:1,000), Top panel: wild-type (WT); Bottom panal: Neurofilament medium chain shRNA knockdown (KD). Nuclei were stained blue with DAPI; Neurofilament medium chain was stained magenta with Alexa Fluor® 647. Scale bar, 20 μm.
Flow cytometric analysis of neurofilament medium chain expression in HeLa cells using neurofilament medium chain antibody (Cat#63480, 1:2,000). Green, isotype control; red, neurofilament medium chain.
