Human TROP-2 Antibody

Detection of Human TROP‑2 by Western Blot. Western blot shows lysates of NCI-N87 human gastric carcinoma cell line. PVDF membrane was probed with 1 µg/mL of Goat Anti-Human TROP-2 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF650) followed by HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF019). A specific band was detected for TROP-2 at approximately 45-50 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 8.

Detection of TROP‑2 in PC‑3 Human Cell Line by Flow Cytometry. PC-3 human prostate cancer cell line was stained with Goat Anti-Human TROP-2 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF650, filled histogram) or control antibody (Catalog # AB-108-C, open histogram), followed by Phycoerythrin-conjugated Anti-Goat IgG Secondary Antibody (Catalog # F0107).

TROP‑2 in Human Brain. TROP-2 was detected in immersion fixed paraffin-embedded sections of human brain (frontal cortex) using Goat Anti-Human TROP-2 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF650) at 10 µg/mL overnight at 4 °C. Before incubation with the primary antibody tissue was subjected to heat-induced epitope retrieval using Antigen Retrieval Reagent-Basic (Catalog # CTS013). Tissue was stained using the Anti-Goat HRP-DAB Cell & Tissue Staining Kit (brown; Catalog # CTS008) and counterstained with hematoxylin (blue). View our protocol for Chromogenic IHC Staining of Paraffin-embedded Tissue Sections.

Detection of Human TROP-2 by Immunohistochemistry Downregulation of Trop2 inhibits cell invasion. (A) The invasive capability of Hep2 cells transfected with NC or Trop2 siRNA (Trop2-S1) was measured at the indicated time points using the Transwell assay. (B) The number of cells on the underside of the chamber was counted. NC, negative control, siRNA, small interfering RNA. Image collected and cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/25779928), licensed under a CC-BY license. Not internally tested by R&D Systems.

Detection of Human TROP-2 by Western Blot Knockdown of Trop2 expression in Hep2 cells by siRNA. (A) Trop2 mRNA expression in Hep2 cells was examined by reverse transcription-quantitative polymerase chain reaction 48 h after transfection with Trop2 siRNA or NC, as indicated (*P<0.05, **P<0.001). (B) Western blot analysis of Trop2 protein expression in Hep2 cells 48 h following transfection with siRNA as indicated. siRNA, small interfering RNA; NC, negative control. Image collected and cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/25779928), licensed under a CC-BY license. Not internally tested by R&D Systems.

Detection of Human TROP-2 by Knockdown Validated Trop2 Loss Promotes ErbB3 Activation in HNSCC CellsA. Immunofluorescence images of Trop2 staining (green) in SCC1 HNSCC cells after stable knockdown using short hairpins targeting the Trop2 cDNA. Nuclei are counterstained with 4′6-diamidino-2-phenylindole (DAPI). B. Results of a phosphorylated receptor tyrosine kinase antibody array demonstrating elevated p-ErbB3 in lysates from Trop2 knockdown SCC1 cells. The exposures were normalized to the control spots (four corners), which exhibit equal intensities. C&D. Representative immunoblots showing hyperactivation of ErbB3 and AKT caused by Trop2 loss in SCC1 and SCC25 HNSCC cells. Two short hairpins targeting distinct regions of the Trop2 cDNA were used. E. Reduction of ErbB3 activity after ectopic expression of an RNAi-resistant Trop2 cDNA in Trop2 knockdown SCC1 cells. Arrow points to the lower band which is the correct size for Trop2. F. Ectopic expression of a Flag-epitope tagged Trop2 cDNA in SCC1 cells suppresses basal ErbB3 and AKT activation. Control is an empty vector. Relative increases in phosphoproteins in control versus experimental groups were quantified by photodensitometry after normalization to total ErbB3 or AKT protein which served as an internal controls. Immunoblots are representative of at least three independent experiments. Significance was measured by student's t test, * (P<0.05), ** (P<0.01), *** (P<0.001). Image collected and cropped by CiteAb from the following publication (https://www.oncotarget.com/lookup/doi/10.18632/oncotarget.2423), licensed under a CC-BY license. Not internally tested by R&D Systems.

Detection of Human TROP-2 by Western Blot Trop2 expression in laryngeal squamous cell carcinoma tissue. (A) Western blot analysis of the protein expression of Trop2 in laryngeal carcinoma tissues (c1, c2, c3 and c4) and paired paracancerous tissues (p1, p2, p3 and p4). Actin was used as a loading control. Representative (B) negative and (C) high expression of Trop2 in paraffin embedding laryngeal carcinoma and precancerous tissues, demonstrated using immunohistochemical staining. Image collected and cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/25779928), licensed under a CC-BY license. Not internally tested by R&D Systems.

Detection of Human TROP-2 by Knockdown Validated Trop2 Loss Promotes ErbB3 Activation in HNSCC CellsA. Immunofluorescence images of Trop2 staining (green) in SCC1 HNSCC cells after stable knockdown using short hairpins targeting the Trop2 cDNA. Nuclei are counterstained with 4′6-diamidino-2-phenylindole (DAPI). B. Results of a phosphorylated receptor tyrosine kinase antibody array demonstrating elevated p-ErbB3 in lysates from Trop2 knockdown SCC1 cells. The exposures were normalized to the control spots (four corners), which exhibit equal intensities. C&D. Representative immunoblots showing hyperactivation of ErbB3 and AKT caused by Trop2 loss in SCC1 and SCC25 HNSCC cells. Two short hairpins targeting distinct regions of the Trop2 cDNA were used. E. Reduction of ErbB3 activity after ectopic expression of an RNAi-resistant Trop2 cDNA in Trop2 knockdown SCC1 cells. Arrow points to the lower band which is the correct size for Trop2. F. Ectopic expression of a Flag-epitope tagged Trop2 cDNA in SCC1 cells suppresses basal ErbB3 and AKT activation. Control is an empty vector. Relative increases in phosphoproteins in control versus experimental groups were quantified by photodensitometry after normalization to total ErbB3 or AKT protein which served as an internal controls. Immunoblots are representative of at least three independent experiments. Significance was measured by student's t test, * (P<0.05), ** (P<0.01), *** (P<0.001). Image collected and cropped by CiteAb from the following publication (https://www.oncotarget.com/lookup/doi/10.18632/oncotarget.2423), licensed under a CC-BY license. Not internally tested by R&D Systems.