Human/Mouse/Rat Ninjurin-2 Antibody

Detection of Ninjurin‑2 in Human Tonsil Ninjurin‑2 was detected in immersion fixed paraffin-embedded sections of Human Tonsil using Sheep Anti-Human/Mouse/Rat Ninjurin‑2 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF5056) at 3 µg/mL for 1 hour at room temperature followed by incubation with the Anti-Sheep IgG VisUCyte™ HRP Polymer Antibody (Catalog # VC006). Before incubation with the primary antibody, tissue was subjected to heat-induced epitope retrieval using VisUCyte Antigen Retrieval Reagent-Basic (Catalog # VCTS021). Tissue was stained using DAB (brown) and counterstained with hematoxylin (blue). Specific staining was localized to cell surface on lymphocytes. View our protocol for IHC Staining with VisUCyte HRP Polymer Detection Reagents.

Detection of Human/Mouse/Rat Ninjurin‑2 by Western Blot. Western blot shows lysates of human thymus, mouse thymus, rat thymus, NTera‑2 human testicular embryonic carcinoma cells and HT‑29 human colon adenocarcinoma cells. PVDF membrane was probed with 1 µg/mL of Sheep Anti-Human/Mouse/Rat Ninjurin‑2 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF5056) followed by HRP-conjugated Anti-Sheep IgG Secondary Antibody (Catalog # HAF016). A specific band was detected for Ninjurin‑2 at approximately ~26kDa kDa (as indicated). This experiment was conducted under reducing conditions and using Western Blot Buffer Group 1.

Detection of Ninjurin-2 by Western Blot NINJ2 in ECF-R regulates chemo-sensitivity and cell cycle arrest. (A) ECF-R MKN-74 cells were transduced with shRNA lentiviral particles targeting human NINJ2 (clone 1 and clone 2) or the pLKO.1-puro empty control. Viability analysis using crystal violet staining (Left) and the CCK-8 assay (Right) 3 weeks after ECF treatment in mock and NINJ2 K/D ECF-R MKN-74 cells. (B) Representative ECF IC50 values for MKN-74 cells overexpressing NINJ2 isoform-1 and isoform-3. (C) (Left) Cell cycle status of mock-, NINJ2 isoform-1-, and isoform-3-O/E MKN-74 cells. (Right) Quantification of the left FACS plots. *p < 0.05 versus mock (S-phase reduction), #p < 0.05 versus mock (G0/G1-phase increase). (D) Western blot analysis of NINJ2, p27KIP1, CDK2, CDK4, CDK6, CDC25a, cyclin D1, cyclin E1, Rb (p-S780), Rb (p-S795), total Rb, and GAPDH in scramble siRNA and NINJ2 K/D MKN-74 cells. K/D, knockdown. Data are presented as mean ± SD Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/40518514), licensed under a CC-BY license. Not internally tested by R&D Systems.

Detection of Ninjurin-2 by Western Blot NINJ2 expression in ECF-R gastric cancer cells. (A) Representative IC50 values for parent (WT) and ECF-resistant (ECF-R) MKN-74 and SNU-484 cells. The IC50 values are given in an equation using a four-parameter logistic curve. (B) Parent and ECF-R MKN-74 cells were individually transplanted into nude mice and left until the tumor reached a volume of 100 mm3. The tumor-bearing mice were given ECF, and the tumor volume was measured. *p < 0.05 versus parent at each time point. (C) (Left) Venn diagram showing top genes with 15-fold changes in the RNA-sequencing of WT and ECF-R genes related to “Integral component of plasma membrane (GO:0005887)” and “Cell adhesion (GO:0007155).” (Right) Heatmap showing the gene expression level associated with resistance and the two gene ontology categories. (D) RGV analysis for the human NINJ2 isoform based on RNA-sequencing of WT and ECF-R (hg19 base) cells. (E) mRNA levels of NINJ2 was analyzed by quantitative real-time PCR in parent and ECF-R cells from the MKN-28/74, MKN-74, MKN-45, SNU-484, SNU-520, and SNU-688 lines. (F) Protein levels of NINJ2 in parent and ECF-R MKN-74 and SNU-484 cells were determined by Western blot analysis. ECF, epirubicin, cisplatin, and 5-fluorouracil; ECF-R, ECF-resistance. Data are presented as mean ± SD Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/40518514), licensed under a CC-BY license. Not internally tested by R&D Systems.