PSAP Antibody [Unconjugated]

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 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.

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.

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 (//pubmed.ncbi.nlm.nih.gov/31864418), licensed under a CC-BY license. Not internally tested by R&D Systems.

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 (//pubmed.ncbi.nlm.nih.gov/31864418), licensed under a CC-BY license. Not internally tested by R&D Systems.

Western blot shows culture media of HeLa human cervical epithelial carcinoma parental cell line and PSAP knockout HeLa cell line (KO). Nitrocellulose membrane was probed with Rabbit Anti-Human PSAP Antigen Affinity-purified Polyclonal Antibody (Catalog # AF8520) followed by HRP-conjugated secondary antibody. A specific band was detected for PSAP at approximately 58.1 kDa (as indicated) in the parental HeLa cell line, but is not detectable in knockout HeLa cell line. Primary antibody dilution used: 1:1000. The Ponceau stained transfer of the blot is shown. This experiment was conducted under reducing conditions. Image, protocol, and testing courtesy of YCharOS Inc. See ycharos.com for additional details.

HeLa human cervical epithelial carcinoma cell line culture medium were prepared and immunoprecipitation was performed using 2.0 μg of Rabbit Anti-Human PSAP Antigen Affinity-purified Polyclonal Antibody (Catalog # AF8520) pre-coupled to Dynabeads Protein A. Immunoprecipitated PSAP was detected in Western Blot with a rabbit PSAP antibody used at 1/1000. The Ponceau stained transfer of the blot is shown. SM=4% starting material; UB=4% unbound fraction; IP=immunoprecipitate; HC=antibody heavy chain. Image, protocol and testing courtesy of YCharOS Inc. (ycharos.com).

• Reactivity: Hu
• Applications: WB, Simple Western, IP, ICC/IF, KO
• Clonality: Polyclonal
• Host: Rabbit
• Conjugate: Unconjugated

PSAP Antibody [Unconjugated] Summary

• Immunogen: S. frugiperda insect ovarian cell line Sf21-derived recombinant human PSAP
  Gly17-Asn524
  Accession # P07602
• Specificity: Detects human PSAP in direct ELISAs and Western blots.
• Source: N/A
• Isotype: IgG
• Clonality: Polyclonal
• Host: Rabbit
• Gene: PSAP
• Purity Statement: Antigen Affinity-purified

Applications/Dilutions

• Dilutions:
  • Immunocytochemistry 1:100 dilution
  • Immunoprecipitation 1 ug/mL
  • Knockout Validated
  • Simple Western 2 ug/mL
  • Western Blot 1:1000 dilution

Packaging, Storage & Formulations

• Storage: Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
  12 months from date of receipt, -20 to -70 degreesC, as supplied. 1 month, 2 to 8 degreesC under sterile conditions after opening. 6 months, -20 to -70 degreesC under sterile conditions after opening.
• Buffer: Supplied as a solution in PBS containing BSA, Glycerol and Sodium Azide. See Certificate of Analysis for details.
• Preservative: Sodium Azide

Notes

* Contains <0.1% Sodium Azide, which is not hazardous at this concentration according to GHS classifications. Refer to SDS for additional information and handling instructions. This product is produced by and ships from R&D Systems, Inc., a Bio-Techne brand.

Alternate Names for PSAP Antibody [Unconjugated]

• FLJ00245
• GLBA
• proactivator polypeptide
• prosaposin
• PSAP
• SAP1
• SAP1MGC110993
• sphingolipid activator protein-1

Background

Prosaposin behaves as a myelinotrophic and neurotrophic factor, whose effects are mediated by its G-protein-coupled receptors, GPR37 and GPR37L1, undergoing ligand-mediated internalization followed by ERK phosphorylation signaling.By similarity Saposins are specific low-molecular mass non-enzymic proteins and participate in the lysosomal degradation of sphingolipids, which takes place by the sequential action of specific hydrolases. The Prosaposin gets cleaved into 5 chains, Saposin A, B, B-Val, C and D. It also contains several disulfide bonds as well as glycosylation sites.

Limitations

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

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We have publications tested in 1 confirmed species: Human.

We have publications tested in 3 applications: IHC-Fr, Immunoprecipitation, Western Blot.

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Showing Publications 1 - 2 of 2.

Publications using AF8520	Applications	Species
Varga, AJ;Nemeth, IB;Kemeny, L;Varga, J;Tiszlavicz, L;Kumar, D;Dodd, S;Simpson, AWM;Buknicz, T;Beynon, R;Simpson, D;Krenacs, T;Dockray, GJ;Varro, A; Elevated Serum Gastrin Is Associated with Melanoma Progression: Putative Role in Increased Migration and Invasion of Melanoma Cells International journal of molecular sciences 2023-11-28 [PMID: 38069171] (Western Blot, Human)	Western Blot	Human
A Mendsaikha, I Tooyama, JP Bellier, GE Serrano, LI Sue, LF Lue, TG Beach, DG Walker Characterization of lysosomal proteins Progranulin and Prosaposin and their interactions in Alzheimer's disease and aged brains: increased levels correlate with neuropathology Acta Neuropathol Commun, 2019-12-21;7(1):215. 2019-12-21 [PMID: 31864418] (IHC-Fr, Immunoprecipitation, Western Blot, Human)	IHC-Fr, Immunoprecipitation, Western Blot	Human

Find general support by application which include: protocols, troubleshooting, illustrated assays, videos and webinars.

• Immunoprecipitation
• Western Blot
• View all Protocols, Troubleshooting, Illustrated assays and Webinars.

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Publications for PSAP Antibody (AF8520)(2)

Publications using AF8520

• Varga, AJ;Nemeth, IB;Kemeny, L;Varga, J;Tiszlavicz, L;Kumar, D;Dodd, S;Simpson, AWM;Buknicz, T;Beynon, R;Simpson, D;Krenacs, T;Dockray, GJ;Varro, A; Elevated Serum Gastrin Is Associated with Melanoma Progression: Putative Role in Increased Migration and Invasion of Melanoma Cells International journal of molecular sciences 2023-11-28 [PMID: 38069171] (Western Blot, Human)
• A Mendsaikha, I Tooyama, JP Bellier, GE Serrano, LI Sue, LF Lue, TG Beach, DG Walker Characterization of lysosomal proteins Progranulin and Prosaposin and their interactions in Alzheimer&#039;s disease and aged brains: increased levels correlate with neuropathology Acta Neuropathol Commun, 2019-12-21;7(1):215. 2019-12-21 [PMID: 31864418] (IHC-Fr, Immunoprecipitation, Western Blot, Human)

Review for PSAP Antibody (AF8520) (1)

Average Rating: 5

(Based on 1 review)

We have 1 review tested in 1 species: Human.

reviewed by: Verified Customer WB Human 04/04/2020

Summary

• Application: Western Blot
• Sample Tested: A549 cells,H441
• Species: Human
• Lot: CJCW0115011

Bioinformatics

• Gene Symbol: PSAP
• Uniprot:
  • P07602()