Human PSAP Antibody

Detection of Human PSAP by Western Blot. Western blot shows lysates of A431 human epithelial carcinoma cell line. PVDF membrane was probed with 1:1000 dilution of Rabbit Anti-Human PSAP Antigen Affinity-purified Polyclonal Antibody (Catalog # AF8520) followed by HRP-conjugated Anti-Rabbit IgG Secondary Antibody (Catalog # HAF008). Specific bands were detected for PSAP at approximately 60-80 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.

PSAP in A431 Human Cell Line. PSAP was detected in immersion fixed A431 human epithelial carcinoma cell line using Rabbit Anti-Human PSAP Antigen Affinity-purified Polyclonal Antibody (Catalog # AF8520) at 1:100 dilution for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Rabbit IgG Secondary Antibody (red; Catalog # NL004) and counterstained with DAPI (blue). Specific staining was localized to Golgi. View our protocol for Fluorescent ICC Staining of Cells on Coverslips.

Detection of Human PSAP by Simple Western^TM. Simple Western lane view shows lysates of A431 human epithelial carcinoma cell line, loaded at 0.2 mg/mL. A specific band was detected for PSAP at approximately 98 kDa (as indicated) using 2 µg/mL of Rabbit Anti-Human PSAP Antigen Affinity-purified Polyclonal Antibody (Catalog # AF8520). This experiment was conducted under reducing conditions and using the 12-230 kDa separation system.

Detection of Human PSAP by Immunocytochemistry/Immunofluorescence Confocal microscopy of prosaposin localization on plaques and different cell types. (a-c). A beta (green) (a) and PSAP (red) plaque (b) with limited colocalization (C – yellow) in an AD case. Scale bar represents 30 μm. (d-f). Comparison of colocalization in plaque of PGRN (green) and A beta (blue) (d) with PSAP (red) and A beta (blue) in triple-stained AD section. Merged images show extensive colocalization of PGRN and PSAP (yellow) but limited overlap with A beta -positive structures. Scale bar represents 30 μm. (G-I). Merged images of PSAP (red) immunoreactivity with microglial markers IBA-1 (g) and CD68 (h) (green) and astrocyte marker GFAP (green) show some expression of PSAP in both cell types (yellow). These images show that PSAP (red) is predominantly in cells with morphology of neurons. Scale bar represents 10 μm. (j-l). Merged images of CD68 (green) and PSAP (red) on plaques in low plaque case (J), high plaque case (k) and AD case (l). Significant amounts of PSAP immunoreactivity (red) can be observed on all plaques but with only limited colocalization with CD68 in infiltrating microglia. Scale bar represents 30 μm. (m-n) Merged images of AT180 (pTau) (green) and PSAP (red) on tangle in low plaque case (M), high plaque case (n) and Alzheimer’s disease case (o). Very limited amounts of PSAP immunoreactivity (yellow) can be observed on tangles. Panel M and N show intracellular tangles with DAPI-positive nuclei, while panel O shows extracellular tangle. Scale bar represents 30 μm. Image collected and cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/31864418), licensed under a CC-BY license. Not internally tested by R&D Systems.

Detection of Human PSAP by Immunocytochemistry/Immunofluorescence Confocal microscopy of prosaposin localization on plaques and different cell types. (a-c). A beta (green) (a) and PSAP (red) plaque (b) with limited colocalization (C – yellow) in an AD case. Scale bar represents 30 μm. (d-f). Comparison of colocalization in plaque of PGRN (green) and A beta (blue) (d) with PSAP (red) and A beta (blue) in triple-stained AD section. Merged images show extensive colocalization of PGRN and PSAP (yellow) but limited overlap with A beta -positive structures. Scale bar represents 30 μm. (G-I). Merged images of PSAP (red) immunoreactivity with microglial markers IBA-1 (g) and CD68 (h) (green) and astrocyte marker GFAP (green) show some expression of PSAP in both cell types (yellow). These images show that PSAP (red) is predominantly in cells with morphology of neurons. Scale bar represents 10 μm. (j-l). Merged images of CD68 (green) and PSAP (red) on plaques in low plaque case (J), high plaque case (k) and AD case (l). Significant amounts of PSAP immunoreactivity (red) can be observed on all plaques but with only limited colocalization with CD68 in infiltrating microglia. Scale bar represents 30 μm. (m-n) Merged images of AT180 (pTau) (green) and PSAP (red) on tangle in low plaque case (M), high plaque case (n) and Alzheimer’s disease case (o). Very limited amounts of PSAP immunoreactivity (yellow) can be observed on tangles. Panel M and N show intracellular tangles with DAPI-positive nuclei, while panel O shows extracellular tangle. Scale bar represents 30 μm. Image collected and cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/31864418), licensed under a CC-BY license. Not internally tested by R&D Systems.