Human/Rat Lactate Dehydrogenase A/LDHA Antibody

Lactate Dehydrogenase A/LDHA in PC‑3 Human Cell Line.

Lactate Dehydrogenase A/LDHA was detected in immersion fixed PC-3 human prostate cancer cell line using Rabbit Anti-Human/Rat Lactate Dehydrogenase A/LDHA Monoclonal Antibody (Catalog # MAB9158) at 3 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Rabbit IgG Secondary Antibody (red; Catalog # NL004) and counterstained with DAPI (blue). Specific staining was localized to cytoplasm. View our protocol for Fluorescent ICC Staining of Cells on Coverslips.

Detection of Lactate Dehydrogenase A/LDHA by Western Blot

Effect of FoxO3a on relevant enzymes involved in carbohydrate metabolism. A double set of cells was transiently transfected in suspension with F3aAAA and pcDNA3 plasmids for 48 h. FoxO3a, PFKP, LDHA, and Aldolase A expressions were assessed at the RNA and protein level in Tam-resistant MCF-7/TR (a–e), T47D/TR (f–j) and ZR-75/TR (k–o). mRNA content was normalized vs. the relative 18S rRNA content, and beta -actin was employed for loading controls (* p < 0.05, ** p < 0.005). Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/38132097), licensed under a CC-BY license. Not internally tested by R&D Systems.

Detection of Lactate Dehydrogenase A/LDHA by Western Blot

Influence of FoxO3a expression on glycolysis of TamR breast cancer cells. TamR/TetOn-AAA and TamR/TetOn-V were plated, left to attach and then treated with Dox 1 µg/mL for 48 h to induce FoxO3a expression. (A) FoxO3a overexpression was determined by WB. GAPDH = loading control. Glucose content (B) LDH (Lactate dehydrogenase) activity in cell lysates (C) and in the growing media (D) were analyzed as described in Section 2. Data are reported as the mean ± SD of three independent experiments (at least in triplicate). Statistical significances are calculated by using Student’s t-test evaluating the differences between TamR/TetOn-AAA and TamR/TetOn-V samples (* p < 0.05). Proteins involved in Carbohydrate Metabolism differentially expressed in the comparison analysis in TamR/TetOn-AAA with respect to TamR/TetOn-V (E). The expression fold changes of reported proteins were evaluated in TamR/TetOn-AAA with respect to TamR/TetOn-V by using Ingenuity Pathway Analysis (Qiagen), fixing the cut-off to 1.6 (F). Proteomic data were confirmed by RNA and WB analysis. To this aim, a duplicate set of cells was treated with Dox 1 µg/mL for 48 h. The RNA (G–J) and protein (K) expressions of FoxO3a, AldoA, LDHA and PFKM were assessed. mRNA content was normalized vs. the relative 18S rRNA content, and beta -actin was employed as a loading control. (L–N) Analysis of genes involved in carbohydrate metabolism that correlated with FOXO3 (cBioPortal dataset) [26], Spearman and Pearson correlation coefficient with the respective p-value are reported. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/38132097), licensed under a CC-BY license. Not internally tested by R&D Systems.

Detection of Lactate Dehydrogenase A/LDHA by Western Blot

Effect of FoxO3a on relevant enzymes involved in carbohydrate metabolism. A double set of cells was transiently transfected in suspension with F3aAAA and pcDNA3 plasmids for 48 h. FoxO3a, PFKP, LDHA, and Aldolase A expressions were assessed at the RNA and protein level in Tam-resistant MCF-7/TR (a–e), T47D/TR (f–j) and ZR-75/TR (k–o). mRNA content was normalized vs. the relative 18S rRNA content, and beta -actin was employed for loading controls (* p < 0.05, ** p < 0.005). Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/38132097), licensed under a CC-BY license. Not internally tested by R&D Systems.