Human/Mouse/Rat PEDFR/PNPLA2 Antibody

Detection of Human and Mouse PEDF R/PNPLA2 by Western Blot. Western blot shows lysates of human adipose tissue and mouse adipose tissue. PVDF membrane was probed with 1 µg/mL of Sheep Anti-Human/Mouse/Rat PEDF R/PNPLA2 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF5365) followed by HRP-conjugated Anti-Sheep IgG Secondary Antibody (Catalog # HAF016). A specific band was detected for PEDF R/PNPLA2 at approximately 55 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 8.

PEDF R/PNPLA2 in Mouse Kidney. PEDF R/PNPLA2 was detected in perfusion fixed frozen sections of mouse kidney using 15 µg/mL Sheep Anti-Human/Mouse/Rat PEDF R/PNPLA2 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF5365) overnight at 4 °C. Tissue was stained with the Anti-Sheep HRP-DAB Cell & Tissue Staining Kit (brown; Catalog # CTS019) and counterstained with hematoxylin (blue). Specific labeling was localized to the plasma membrane of epithelial cells in tubules. View our protocol for Chromogenic IHC Staining of Frozen Tissue Sections.

Detection of Mouse PEDF R/PNPLA2 by Simple Western^TM. Simple Western lane view shows lysates of mouse adipose tissue, loaded at 0.2 mg/mL. A specific band was detected for PEDF R/PNPLA2 at approximately 59 kDa (as indicated) using 10 µg/mL of Sheep Anti-Human/Mouse/Rat PEDF R/PNPLA2 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF5365) followed by 1:50 dilution of HRP-conjugated Anti-Sheep IgG Secondary Antibody (Catalog # HAF016). This experiment was conducted under reducing conditions and using the 12-230 kDa separation system.

Detection of Human PEDFR/PNPLA2/ATGL by Western Blot Loss-of-function analyses of PEDF in Bkid cells. (A) PEDF micro RNA (miR) expressing vector was transfected into Bkid cells. Immunoblot by anti-PEDF antibody showed the reduction of PEDF proteins by miRs. (B) The renal and the hepatic lesions of the results of the intracardiac injections of Bkid cells or Bkid+PEDFmiR cells. The adrenal metastasis was observed. Also, by Bkid cell injections, the renal metastasis was observed frequently (left, 12-positive/12), while no renal nodule was observed in Bkid+PEDFmiR injected mouse (right, 17-negative/18). The ruler is 1mm square. (C) The hepatic metastases of both cells were not inhibited with or without PEDF. (D) The section Hematoxylin/Eosin (H&E) staining of the similar samples to (B). The adrenal hypertrophy was observed both in Bkid- and Bkid+PEDFmiR-injected mouse organs. The renal metastatic lesion (met) was seen as the lighter color. The scale is 2 mm. (E) The hepatic metastasis. Both tumors had the cavity. (F) PEDF-immunohistochemistry (IHC) of the section (D). PEDF staining was observed inside of the adrenal gland cystic vesicle of Bkid tumors. The overall staining level of PEDF in Bkid tumor was higher than Bkid+PEDFmiR cells. The scale is 1 mm. (G) IHC for PEDF on paraffin sections of collagen gel-embedded 143B and 143B+PEDF cells. (H-L) The renal section IHC by various antibodies. (H) Anti-SPARC (osteonectin)-IHC indicates that the lesions are derived from the injected osteosarcoma. In the section of kidney from Bkid-injected mouse, there is SPARC-positive tumor both in kidney and adrenal gland sides. On the right, the kidney from Bkid+PEDFmiR-injected mouse had SPARC-positive cells only in adrenal gland. The scale indicates 100 µm. (I–L) The scale indicates 20 µm. (I) The higher magnified picture of the tissue surrounded glomerulus. There is SPARC-positive cells were seen around the glomerulus of Bkid-injected mouse (arrow), while no signal was seen in Bkid+PEDFmiR-injected mouse, (J) PEDF-IHC. PEDF was accumulated in the podocytes around the glomerulus. (K) 67kDa laminin receptor (lamR)-IHC. lamR is a potential PEDF receptor. Arrows indicated lamR signals in the podocytes around and the vein in the glomerulus. (L) CD31-IHC in glomerulus. (M, N) The liver section IHCs with CD31 antibody. The vascular formation was observed. (M) No closed blood vessel was seen in Bkid tumor (above). Near the tumor-liver boundary in Bkid sample (below). (N) The blood vessels could be seen as the closed oval form in Bkid+PEDFmiR tumor (above). Tumor-liver boundary of Bkid+PEDFmiR (below). There were a lot of the closed blood vessels (arrowheads). (O) The closed shapes of CD31-staining were counted only in the tumor regions, and the results were normalized by CD31-positive area. PEDF expression severely disturbed the formation of blood vessels (p=0.015). ad, adrenal gland; kid, kidney; met, metastatic lesion; liv, liver. Image collected and cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/35174090), licensed under a CC-BY license. Not internally tested by R&D Systems.