Journal: Journal of Oncology

Article Title: High Aldehyde Dehydrogenase Levels Are Detectable in the Serum of Patients with Lung Cancer and May Be Exploited as Screening Biomarkers

doi: 10.1155/2019/8970645

Figure Lengend Snippet: Western blot analysis of the polyclonal anti-RTNKEASI serum in A549 cell line. SDS gel was loaded with protein extract from non-small cell lung cancer A549 cell line followed by electrophoresis and immunoblotting. The blots were incubated with the preimmune serum and the polyclonal anti-RTNKEASI serum produced in rabbits from Rockland (a) or BioGenes (b). The 55 kDa target of anti-RTNKEASI appears only in sera of animals after immunization, and it is absent in the presera from the same rabbit. These results were confirmed with both Rockland and BioGenes antibodies. A PCA was performed by incubating the corresponding blots with the polyclonal anti-RTNKEASI serum from Rockland, pretreated with or without the peptide RTNKEASI (c) and also β -actin, as endogenous control. The intensity of the band approximately at 55 kDa detected by the secondary anti-rabbit horseradish peroxidase-conjugated was reduced after the preincubation of the antibody with the peptide RTNKEASI.

Article Snippet: The production of two different rabbit polyclonal anti-RTNKEASI immune sera and the synthesis of TLP-derived peptide RTNKEASI [ ] were both conducted at Rockland Immunochemicals Inc. (Gilbertsville, PA, USA) and at BioGenes GmbH (Berlin, Germany).

Techniques: Western Blot, SDS-Gel, Electrophoresis, Incubation, Produced, Control

Journal: Journal of Oncology

Article Title: High Aldehyde Dehydrogenase Levels Are Detectable in the Serum of Patients with Lung Cancer and May Be Exploited as Screening Biomarkers

doi: 10.1155/2019/8970645

Figure Lengend Snippet: Western blot analysis for the polyclonal anti-RTNKEASI serum in cancer cell lines and cellular supernatant. (a) Protein extracts from non-small cell lung cancer A549, normal lung tissue MRC-5, Burkitt lymphoma CA46, leukemia HL60, breast cancer MCF-7, cervical carcinoma Hela, and prostate cancer PC-3 cell lines were loaded into SDS gel and subjected to electrophoresis and immunoblotting with the polyclonal anti-RTNKEASI serum from Rockland. High 55 kDa protein levels were identified in A549 cells and a lower amount in Hela, PC-3, CA46 and HL-60 cell lines whereas no protein levels were observed in MCF-7 cell line from breast cancer and in MRC-5 from normal lung tissue. An anti-ß-actin antibody was used for a loading control. (b) Protein extracts from cellular supernatant of non-small cell lung cancer A549 and cell lysate from A549 and MRC-5 cells were loaded into SDS gel and subjected to electrophoresis and immunoblotting with the polyclonal anti-RTNKEASI serum from Rockland. A specific band at 55 kDa was detected in the cellular supernatant sample with respect to cell lysate from A549; MRC-5 was used as negative control.

Article Snippet: The production of two different rabbit polyclonal anti-RTNKEASI immune sera and the synthesis of TLP-derived peptide RTNKEASI [ ] were both conducted at Rockland Immunochemicals Inc. (Gilbertsville, PA, USA) and at BioGenes GmbH (Berlin, Germany).

Techniques: Western Blot, SDS-Gel, Electrophoresis, Control, Negative Control

Journal: Journal of Oncology

Article Title: High Aldehyde Dehydrogenase Levels Are Detectable in the Serum of Patients with Lung Cancer and May Be Exploited as Screening Biomarkers

doi: 10.1155/2019/8970645

Figure Lengend Snippet: 2D gel electrophoresis and western blot analysis with cell lysates from A549 and MCF-7 cell lines using the polyclonal anti-RTNKEASI serum and preimmune serum. Protein extract from A549 and MCF-7 cells were loaded in 2D gel followed by electrophoresis and transferred to PVDF for immunoblotting with the polyclonal anti-RTNKEASI serum or preserum. A rectangle marks the location of the ALDHs detected with the polyclonal anti-RTNKEASI serum in A549 cells; the reactive spots indicated with arrows were absent in MCF-7 cells and in the blot incubated with the preserum. These protein spots at 55 kDa, excited and subjected to MS analysis, correspond to ALDH1A1.

Article Snippet: The production of two different rabbit polyclonal anti-RTNKEASI immune sera and the synthesis of TLP-derived peptide RTNKEASI [ ] were both conducted at Rockland Immunochemicals Inc. (Gilbertsville, PA, USA) and at BioGenes GmbH (Berlin, Germany).

Techniques: Two-Dimensional Gel Electrophoresis, Electrophoresis, Western Blot, Incubation