Introduction: Pancreatobiliary lymphadenopathy (PBL) may be due to a number of benign or malignant causes. Tissue sampling of these lymph nodes (LN) can be possible with the help of endoscopic ultrasound (EUS). Aim of this study was to identify the etiology of the PBL, morphology, and factors predicting good yield of biopsy with EUS. Materials and methods: All patients found to have pancreatobiliary lymph node (PBLN) enlargement (>10 mm) on abdominal imaging and referred for EUS-guided biopsy were included in this prospective observational study. The facility of rapid on-site evaluation (ROSE) was not available. Adequacy of the tissue specimen was assessed by the endoscopist with macroscopic on-site evaluation (MOSE) and then sent to histopathologist for final diagnosis. Factors predicting good yield of biopsy were then analyzed. Results: Of the total 87 patients with PBL, 54 (62.1%) were males. Mean age of the patients was 52.0 (±13.4) and range 18–80 years. The commonest locations of PBL were porta hepatis 37 (42.5%), peripancreatic 24 (27.6%), celiac 16 (18.4%), and others 10 (11.5%). Histological reports showed: neoplastic tissue in 34 (39.1%), non-neoplastic in 20 (23%), normal lymphoid tissue (27.6%) and suboptimal in 9 (10.3%). Among the 34 neoplastic causes, 26 had metastatic adenocarcinoma, 5 had lymphoma, and 3 had metastatic neuroendocrine tumors. Among the 20 non-neoplastic causes, 10 had tuberculosis, 4 had anthracosis, and 6 had other findings. Factors predicting good yield of biopsy were a PBLN size ≥12 mm and satisfactory MOSE on both univariate [PBLN (p = 0.005); MOSE (p < 0.0001)] and multivariate [PBLN (p = 0.011); MOSE (p < 0.0001)] analysis. Conclusion: The commonest etiology of PBLN enlargement was metastatic adenocarcinoma among the neoplastic causes and tuberculosis among the non-neoplastic causes. The most common PBLNs approached by EUS were in portahepatis and peripancreatic regions. A good biopsy yield can be predicted with PBLN size of ≥12 mm and a satisfactory MOSE.