Human Cathepsin C/DPPI Antibody

Detection of Human Cathepsin C/DPPI by Western Blot. Western blot shows lysates of A549 human lung carcinoma cell line. PVDF membrane was probed with 1 µg/mL of Goat Anti-Human Cathepsin C/DPPI Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1071) followed by HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF019). A specific band was detected for Cathepsin C/DPPI at approximately 55 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.

Detection of Human Cathepsin C/DPPI by Simple Western^TM. Simple Western lane view shows lysates of A549 human lung carcinoma cell line, loaded at 0.2 mg/mL. A specific band was detected for Cathepsin C/DPPI at approximately 56 kDa (as indicated) using 50 µg/mL of Goat Anti-Human Cathepsin C/DPPI Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1071) followed by 1:50 dilution of HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF109). This experiment was conducted under reducing conditions and using the 12-230 kDa separation system. Non-specific interaction with the 230 kDa Simple Western standard may be seen with this antibody.

Cathepsin C/DPPI in Human Lung Cancer Tissue. Cathepsin C/DPPI was detected in immersion fixed paraffin-embedded sections of human lung cancer tissue using Goat Anti-Human Cathepsin C/DPPI Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1071) at 1 µg/mL overnight at 4 °C. Tissue was stained using the Anti-Goat HRP-DAB Cell & Tissue Staining Kit (brown; Catalog # CTS008) and counterstained with hematoxylin (blue). Specific staining was localized to cytoplasm in cancer cells. View our protocol for Chromogenic IHC Staining of Paraffin-embedded Tissue Sections.

Detection of Human Cathepsin C/DPPI by Western Blot Hypoglycosylation of proteins in knockout HEK293 cell lines.Cell lines were transfected with expression vectors for the following human glycoproteins: (a) SHBG, (b) SHBG derivatives (see diagram) with single glycosylation sites (SHBG N380Q and N396Q), (c) a cathepsin C derivative with a single glycosylation site (CatC delta 234-HA), (d) prosaposin (pSAP-DDKHis), (e) haptoglobin (Hp-DDKHis), (f) hemopexin (Hpx-DDKHis). Endogenous progranulin (pGran) (d) was analyzed using non-transfected cells. The cells were pulse labeled for 10 min and chased for 10 min (pSAP, pGran and CatC delta 234-HA) or pulse labeled for 5 min and chased for 20 min (Hp, SHBG and Hpx). Glycoproteins precipitated with anti-DDK, anti-HA or anti-SHBG were resolved by PAGE in SDS. EH designates treatment with endoglycosidase H. Quantified values below gel lanes (a–f) are for the displayed image, which is representative of two or more experiments. (b) The vertical line indicates the excision of three intervening gel lanes. (d) Resolution of pGran glycoforms is not sufficient for quantification. Nonglycosylated forms of pSAP, Hp, and Hpx that comigrate with the EH-digested form of the substrate in HEK293 cells correspond to a non-translocated precursor and were not used to calculate the average number of glycans. (a–f) Phosphorimages were cropped to display the region of interest. Full-length phosphorimages are shown in Supplemental Fig. S5. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/26864433), licensed under a CC-BY license. Not internally tested by R&D Systems.