Journal: Cell communication and signaling : CCS

Article Title: Peritoneal cavity-derived small extracellular vesicles from aged tumor-naïve hosts promote ovarian cancer adhesion and invasion.

doi: 10.1186/s12964-025-02273-1

Figure Lengend Snippet: Fig. 6 Blocking sEV-associated proteins abrogates the enhanced meso-mimetic adhesion observed with aged sEVs. sEVs (5 × 107) purified from perito neal lavage obtained from aged hosts or control (PBS) were incubated with function-blocking antibodies directed against (A) β1 integrin (2 µg), (B) CA125 (MUC16, 1 µg) or (C) LYN kinase (1 µg) in a total volume of 200 ul for 3 h prior to adding to OvCa cells for 24 h. The meso-mimetic adhesion assay was then performed as described in Fig. 3. (D) sEVs (5 × 107) purified from peritoneal lavage obtained from aged hosts or control (PBS) were incubated with the Lyn kinase inhibitor TL0259 (0.1 nM) for 3 h prior to adding to OvCa cells for 24 h. The meso-mimetic adhesion assay was then performed as described in Fig. 3. Assays were performed in triplicate. Data were analyzed using Kruskal-Wallis test and Dunn’s multi-comparison test

Article Snippet: Gels were transferred to a polyvinylidene difluoride membrane (ImmobilonP, Millipore) using a Bio-Rad Trans-Blot SD Semi-Dry Transfer Cell device, and blocked in 5% milk in TBST buffer (25 mmol/l Tris pH 7.5, 150 mmol/l NaCl, 0.1% Tween 20) for 1 h at room temperature (RT), then were incubated overnight with antibodies to CD9, CD63, CD81, TSG101, Annexin A5, Integrin β1, integrin α3, filaggrin, transglutaminase2, Lyn, Mhc1 (Santa Cruz Biotechnology), Integrin α2 (Advanced Cellular Biology), and MUC16/CA125 (Dako) at a 1:100 dilution in 5% milk in TBST at 4 °C with gentle rocking.

Techniques: Blocking Assay, Purification, Control, Incubation, Cell Adhesion Assay, Comparison

Journal: Scientific Reports

Article Title: Development and application of two novel monoclonal antibodies against overexpressed CD26 and integrin α3 in human pancreatic cancer

doi: 10.1038/s41598-019-57287-w

Figure Lengend Snippet: Immunoprecipitation and immunodetection by Western blot of ( A ) integrin α3 and ( B ) CD26 antigen with novel mAbs KU44.22B and KU44.13A using lysates from CaOV-3 ovarian cancer cells and AsPC-1 pancreatic cancer cells respectively. Left panel: Immunoprecipitation was performed with novel mAbs ( A ) KU44.22B and ( B ) KU44.13A (5 µg) using sheep anti-mouse dynabeads. Protein bands around ~140 KDa and ~ 260KDa were immunoprecipitated with mAb KU44.22B (A; left panel) and ~110 KDa by mAb KU44.13A (B; left panel) respectively and stained with SimplyBlue™ SafeStain. The ~50/25 KDa bands represent heavy and light chains of the anti-mouse antibody. *( B ) left panel corresponds to a cropped gel; vertically sliced images of juxtaposed lanes that were non-adjacent in the gel have a clear separation delineating the boundary between the gels. Middle panel: Integrin α3 and CD26 antigen were immunoprecipitated with mAbs ( A ) KU44.22B and ( B ) KU44.13A (5 µg) respectively, and probed with the same antibody (30 µg/ml). Target antigens were not immunodetected with either of the mAbs. Right panel: Integrin α3 and CD26 antigen were immunoprecipitated with mAbs ( A ) KU44.22B and ( B ) KU44.13A respectively (5 µg) or commercial anti-integrin α3 and anti-CD26 antibodies (2 µg) and immunodetected with commercial mAbs sc-374242 and ab89398 as described in Methods. Immunodetection of target antigens immunoprecipitated by novel mAbs and probed with commercial mAbs confirmed the target identity. MW: molecular weight marker.

Article Snippet: In conclusion, in this study, we reported the production of two novel monoclonal antibodies against integrin α3 and CD26 on human pancreatic cancer cells, using the liver metastasis pancreatic cancer cell line CFPAC-1 as immunogen.

Techniques: Immunoprecipitation, Immunodetection, Western Blot, Staining, Molecular Weight, Marker

Journal: Scientific Reports

Article Title: Development and application of two novel monoclonal antibodies against overexpressed CD26 and integrin α3 in human pancreatic cancer

doi: 10.1038/s41598-019-57287-w

Figure Lengend Snippet: Identification of proteins recognised by novel mAbs KU44.13A and KU44.22B by mass spectrometry.

Article Snippet: In conclusion, in this study, we reported the production of two novel monoclonal antibodies against integrin α3 and CD26 on human pancreatic cancer cells, using the liver metastasis pancreatic cancer cell line CFPAC-1 as immunogen.

Techniques: