Human PSAP Antibody

R&D Systems | Catalog # AF8520

R&D Systems
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Key Product Details

Validated by

Knockout/Knockdown

Species Reactivity

Validated:

Human

Cited:

Human

Applications

Validated:

Knockout Validated, Western Blot, Immunocytochemistry, Simple Western, Immunoprecipitation

Cited:

Immunohistochemistry-Frozen, Western Blot, Immunoprecipitation

Label

Unconjugated

Antibody Source

Polyclonal Rabbit IgG
View all PSAP Primary Antibodies »

Product Specifications

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. 

Clonality

Polyclonal

Host

Rabbit

Isotype

IgG

Scientific Data Images for Human PSAP Antibody

Detection of Human PSAP antibody by Western Blot.

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 antibody in A431 Human Cell Line by Immunocytochemistry (ICC).

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 Immunocytochemistry/Immunofluorescence

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 Simple WesternTM.

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

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.
Western Blot Shows PSAP Specificity Using Knockout Cell Line.

Western Blot Shows PSAP Specificity Using Knockout Cell Line.

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.
Detection of PSAP by Immunoprecipitation.

Detection of PSAP by Immunoprecipitation.

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).

Applications for Human PSAP Antibody

Application
Recommended Usage

Immunocytochemistry

1:100 dilution
Sample: Immersion fixed A431 human epithelial carcinoma cell line

Immunoprecipitation

1 µg/mL
Sample: Culture medium of HeLa human cervical epithelial carcinoma cell line

Knockout Validated

PSAP is specifically detected in HeLa human cervical epithelial carcinoma parental cell line but is not detectable in knockout HeLa human cervical epithelial carcinoma cell line.

Simple Western

2 µg/mL
Sample: A431 human epithelial carcinoma cell line

Western Blot

1:1000 dilution
Sample: A431 human epithelial carcinoma cell line

Reviewed Applications

Read 2 reviews rated 5 using AF8520 in the following applications:

Formulation, Preparation, and Storage

Purification

Antigen Affinity-purified

Formulation

Supplied as a solution in PBS containing BSA, Glycerol and Sodium Azide. See Certificate of Analysis for details.

Shipping

The product is shipped with polar packs. Upon receipt, store it immediately at the temperature recommended below.

Stability & Storage

Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
  • 12 months from date of receipt, -20 to -70 °C, as supplied.
  • 1 month, 2 to 8 °C under sterile conditions after opening.
  • 6 months, -20 to -70 °C under sterile conditions after opening.

Background: PSAP

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.

Long Name

Prosaposin

Alternate Names

GLBA, SAP1

Entrez Gene IDs

5660 (Human); 19156 (Mouse); 25524 (Rat)

Gene Symbol

PSAP

UniProt

P07602

Additional PSAP Products

Product Documents for Human PSAP Antibody

Certificate of Analysis

To download a Certificate of Analysis, please enter a lot or batch number in the search box below.

Note: Certificate of Analysis not available for kit components.

Download SDS

Product Specific Notices for Human PSAP Antibody

* 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.

For research use only

Citations for Human PSAP Antibody

Customer Reviews for Human PSAP Antibody (2)

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Customer Images

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  • Human PSAP Antibody
    Name: Anonymous
    Application: Western Blot
    Sample Tested: A549 cells and H441
    Species: Human
    Verified Customer | Posted 04/04/2020
    Human PSAP Antibody AF8520
  • Human PSAP Antibody
    Name: Anonymous
    Application: Immunocytochemistry/Immunofluorescence
    Sample Tested: A431 human epithelial carcinoma cell line
    Species: Human
    Verified Customer | Posted 10/28/2018
    Human PSAP Antibody AF8520

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Protocols

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