Human Cathepsin L Antibody

Detection of Recombinant Human Cathepsin L by Western Blot.

Western blot shows 10 ng of Recombinant Human Cathepsin L (952-CY), Recombinant Mouse Cathepsin L (1515-CY), Recombinant Human Cathepsin V (1080-CY), Recombinant Human Cathepsin K, Recombinant Human Cathepsin S (1183-CY), Recombinant Human Cathepsin H (7516-CY), and Recombinant Human Cathepsin F. PVDF Membrane was probed with 0.1 µg/mL of Goat Anti-Human Cathepsin L Antigen Affinity-purified Polyclonal Antibody (Catalog # AF952) followed by HRP-conjugated Anti-Goat IgG Secondary Antibody (HAF109). A specific band was detected for Cathepsin L at approximately 35 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 3.

Cathepsin L in Human Kidney.

Cathepsin L was detected in immersion fixed paraffin-embedded sections of human kidney using Goat Anti-Human Cathepsin L Antigen Affinity-purified Polyclonal Antibody (Catalog # AF952) at 15 µg/mL overnight at 4 °C. Tissue was stained using the Anti-Goat HRP-DAB Cell & Tissue Staining Kit (brown; CTS008) and counterstained with hematoxylin (blue). Specific staining was localized to convoluted tubules. View our protocol for Chromogenic IHC Staining of Paraffin-embedded Tissue Sections.

Detection of Human Cathepsin L by Western Blot

Cathepsins are reduced in GNPTAB- knockout cells, but restored upon GNPTAB reconstitution. a Immunoblotting of lysates from parental, knockout and reconstituted cells for CatB and CatL. The migration of a 30 kDa molecular mass marker is shown to the right. b CatB activity in lysates from parental HAP1, or knockout GNPTAB- or NPC1-cells. Cells were treated with cathepsin B inhibitor (Bi), cathepsin L inhibitor (Li), at 10 or 1 µM, or DMSO vehicle control for 1 h at 37 °C, before lysis and incubation with fluorescent CatB peptide substrate. After 1 h incubation at room temperature, fluorescence was measured. Data represent the mean ± s.d. of three technical replicates. A representative of three independent experiments is shown. c CatB activity in lysates from reconstituted cells. Lysates from transduced HAP1 or GNPTAB− cells were generated and tested for CatB activity, as for panel b. Data represent the mean ± s.d. of three technical replicates. A representative of two independent experiments is shown. For both panels b and c, statistical analysis was performed with a two-tailed Student’s t-test with significance shown as **** P ≤ 0.0001. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/30655525), licensed under a CC-BY license. Not internally tested by R&D Systems.