Mouse Klotho Antibody

Detection of Mouse Klotho by Western Blot. Western blot shows lysates of mouse kidney tissue. PVDF membrane was probed with 0.2 µg/mL of Goat Anti-Mouse Klotho Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1819) followed by HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF109). A specific band was detected for Klotho at approximately 145 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.

Klotho in Mouse Kidney. Klotho was detected in perfusion fixed frozen sections of mouse kidney using 1.7 µg/mL Goat Anti-Mouse Klotho Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1819) overnight at 4 °C. Tissue was stained with the Anti-Goat HRP-DAB Cell & Tissue Staining Kit (brown; Catalog # CTS008) and counterstained with hematoxylin (blue). View our protocol for Chromogenic IHC Staining of Frozen Tissue Sections.

Mouse Klotho ELISA Standard Curve. Recombinant Mouse Klotho protein was serially diluted 2-fold and captured by Rat Anti-Mouse Klotho Monoclonal Antibody (Catalog # MAB1819) coated on a Clear Polystyrene Microplate (Catalog # DY990). Goat Anti-Mouse Klotho Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1819) was biotinylated and incubated with the protein captured on the plate. Detection of the standard curve was achieved by incubating Streptavidin-HRP (Catalog # DY998) followed by Substrate Solution (Catalog # DY999) and stopping the enzymatic reaction with Stop Solution (Catalog # DY994).

Detection of Human Klotho by Western Blot Ectopic expression of Klotho protects renal mitochondrial homeostasis and inhibits tubular senescence in an accelerated aging mouse model. (a) Renal expression of PGC‐1 alpha mRNA in different groups was assessed by real‐time PCR. *p < .05 versus control mice; †p < .05 versus d‐gal‐treated mice (n = 5–6). (b) Representative micrographs show renal expression of PGC‐1 alpha. Paraffin sections were immunostained for PGC‐1 alpha. Arrows indicate positive staining. Scale bar, 50 µm. (c–h) Representative (c) Western blots and graphical representations of (d) phospho‐PGC‐1 alpha, (e) TFAM, (f) COX1, (g) Cytb, and (h) TOMM20 protein expression levels in different groups are presented. *p < .05 versus control mice; †p < .05 versus d‐gal‐treated mice (n = 5–6). (i) Quantification of renal mitochondrial mass is shown. Mitochondrial mass was determined by the fluorescence intensity of MitoTracker deep red staining normalized to DAPI. *p < .05 versus control mice; †p < .05 versus d‐gal‐treated mice (n = 5–6). (j) Representative micrographs show mitochondrial ROS production and ultrastructure. They were assessed by mitoSOX staining and electron microscopy analyses, respectively. The administration of d‐gal induced mitochondrial ROS production, mitochondrial swelling, and cristae disorganization in renal tubular cells. Arrows indicate positive staining. Ectopic expression of Klotho inhibited mitochondrial ROS production and preserved normal structure of mitochondria. Scale bar in mitoSOX staining, 50 μm. Scale bar in TEM, 1 μm. TEM, transmission electron microscopy. (k) Representative staining micrographs show renal expression of p16INK4A, SA‐ beta ‐gal activity, and gamma H2AX. Paraffin kidney sections were stained with antibodies against p16INK4A and gamma H2AX. Frozen kidney sections were stained for SA‐ beta ‐gal activity. Arrows indicate positive staining. Scale bar, 25 μm. (l–n) Representative (l) Western blots and graphical representations of (m) p16INK4A and (n) p19ARF protein expression levels in kidneys are presented. *p < .05 versus control mice; †p < .05 versus d‐gal‐treated mice (n = 5–6) Image collected and cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/31318148), licensed under a CC-BY license. Not internally tested by R&D Systems.