Human Park7/DJ-1 Antibody

Western Blot Shows Human Park7/DJ‑1 Specificity by Using Knockout Cell Line.

Western blot shows lysates of HEK293T human embryonic kidney parental cell line and Park7 knockout HEK293T cell line (KO). PVDF membrane was probed with 0.2 µg/mL of Goat Anti-Human Park7/DJ-1 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF3995) followed by HRP-conjugated Anti-Goat IgG Secondary Antibody (HAF017). A specific band was detected for Park7/DJ-1 at approximately 23 kDa (as indicated) in the parental HEK293T cell line, but is not detectable in knockout HEK293T cell line. GAPDH (AF5718) is shown as a loading control. This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.

Detection of Park7/DJ‑1 in Human Brain Hypothalamus.

Park7/DJ‑1 was detected in immersion fixed paraffin-embedded sections of Human Brain Hypothalamus using Goat Anti-Human Park7/DJ‑1 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF3995) at 1.7 µg/mL for 1 hour at room temperature followed by incubation with the Anti-Goat IgG VisUCyte™ HRP Polymer Antibody (Catalog # VC004). 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 cytoplasm in neurons. View our protocol for IHC Staining with VisUCyte HRP Polymer Detection Reagents.

Detection of Park7/DJ-1 by Western Blot

Intracellular PCSK9 content in Caco-2 cells decreases with cell maturation. Differentiated Caco2 cells at d8 and at d16 were recovered after a 2-h secretion assay, and intracellular proteins were extracted from cells. A: Intracellular PCSK9 content (ng extracted from cells) in samples from six independent experiments were measured by ELISA. B: Intracellular pro-PCSK9, PCSK9 and beta -actin contents were analyzed by WB. Representative images from three independent experiments are shown (left panel). Pro-PCSK9 and PCSK9 contents relative to b-actin content from seven independent experiments are shown (right panel). C: Cells lysates were denatured and incubated 3 h at 37°C in presence or not of endoH or PNGase F enzymes, followed by WB for PCSK9. Bands corresponding to undigested Pro-PCSK9 (Pro-PCSK9), undigested PCSK9 (PCSK9), digested PCSK9 (Dig-PCSK9), and digested Pro-PCSK9 (Dig-Pro-PCSK9) by enzymes are indicated. One representative result from two independent experiments is shown. Histograms represents the median ± interquartile. ∗P value< 0.05, ∗∗∗P value <0.001. PCSK9, proprotein convertase subtilisin/kexin type 9; WB, Western blot. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/34280453), licensed under a CC-BY license. Not internally tested by R&D Systems.