Neprilysin/CD10 Antibody (715820) [Unconjugated]

Neprilysin/CD10 was detected in immersion fixed paraffin-embedded sections of human Kidney using Mouse Anti-Human Neprilysin/CD10, Monoclonal Antibody (Catalog # MAB11821) at 0.25ug/mL at 37 ° Celsius for 2 minutes. Before incubation with the primary antibody, tissue underwent an all-in-one dewaxing and antigen retrieval preprocessing using PreTreatment Module (PT Module) and Dewax and HIER Buffer H (pH 9; Epredia Catalog # TA-999-DHBH). Tissue was stained using the Alexa Fluor™ 555 Goat anti-Mouse IgG Secondary Antibody at 1:100 at 37 ° Celsius for 2 minutes. (Yellow; Lunaphore Catalog # DR555MS) and counterstained with DAPI (blue; Lunaphore Catalog # DR100). Specific staining was localized to the membrane. Protocol available in COMET™ Panel Builder.​

Western blot shows lysates of Daudi human Burkitt's lymphoma cell line and Ramos human Burkitt's lymphoma cell line. PVDF membrane was probed with 0.1 µg/mL of Mouse Anti-Human Neprilysin/CD10 Monoclonal Antibody (Catalog # MAB11821) followed by HRP-conjugated Anti-Mouse IgG Secondary Antibody (Catalog # HAF007). A specific band was detected for Neprilysin/CD10 at approximately 100 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.

Neprilysin/CD10 was detected in immersion fixed paraffin-embedded sections of human kidney using Mouse Anti-Human Neprilysin/CD10 Monoclonal Antibody (Catalog # MAB11821) at 0.5 µg/mL for 1 hour at room temperature followed by incubation with the Anti-Mouse IgG VisUCyte™ HRP Polymer Antibody (Catalog # VC001). Tissue was stained using DAB (brown) and counterstained with hematoxylin (blue). Specific staining was localized to convoluted tubules and glomeruli. View our protocol for IHC Staining with VisUCyte HRP Polymer Detection Reagents.

Western blot analysis and quantitative densitometry for NEP levels in RA-differentiated SH-SY5Y cells.The cells were incubated with 0.5% DMSO (control) or 5 μM curcumin (no. 1) or compound 7 or 8 for 24 h, then NEP protein in the cell membrane fraction was measured by western blotting. (a) Western blotting results of three independent experiments. GAPDH was used as the loading control. (b) The quantification of NEP protein levels by Image J. The NEP intensities were normalized to the GAPDH intensities for three independent experiments. Data are presented as the mean ± SD; ns, not significant; **p < 0.01 compared to the DMSO-treated control group by Student’s t-test. Image collected and cropped by CiteAb from the following open publication (//pubmed.ncbi.nlm.nih.gov/27407064), licensed under a CC-BY license. Not internally tested by R&D Systems.

• Reactivity: Hu
• Applications: WB, IHC, mIF
• Clone: 715820
• Clonality: Monoclonal
• Host: Mouse
• Conjugate: Unconjugated
• Concentration: LYOPH

Neprilysin/CD10 Antibody (715820) [Unconjugated] Summary

• Immunogen: Chinese hamster ovary cell line CHO-derived recombinant human Neprilysin/CD10
  Tyr52-Trp750
  Accession # P08473
• Specificity: Detects human Neprilysin/CD10 in direct ELISAs and Western blots. In direct ELISAs, less than 5% cross-reactivity with recombinant mouse (rm) Neprilysin is observed, and no cross-reactivity with rmKell, recombinant human (rh) ECE-1, rhECE-2, or rhNeprilysin-2 is observed.
• Source: N/A
• Isotype: IgG2b
• Clonality: Monoclonal
• Host: Mouse
• Gene: MME
• Purity Statement: Protein A or G purified from hybridoma culture supernatant

Applications/Dilutions

• Dilutions:
  • Immunohistochemistry 0.5-25 ug/mL
  • Multiplex Immunofluorescence 0.25 ug/mL
  • Western Blot 0.1 ug/mL

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 reconstitution. 6 months, -20 to -70 degreesC under sterile conditions after reconstitution.
• Buffer: Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. See Certificate of Analysis for details. *Small pack size (-SP) is supplied either lyophilized or as a 0.2 µm filtered solution in PBS.
• Preservative: No Preservative
• Concentration: LYOPH
• Reconstitution Instructions: Sterile PBS to a final concentration of 0.5 mg/mL. For liquid material, refer to CoA for concentration.

Notes

This product is produced by and ships from R&D Systems, Inc., a Bio-Techne brand.

Alternate Names for Neprilysin/CD10 Antibody (715820) [Unconjugated]

• Atriopeptidase
• CALLA
• CALLAmembrane metallo-endopeptidase (neutral endopeptidase, enkephalinase)
• CD10 antigen
• CD10
• CD10)
• CD10membrane metallo-endopeptidase variant 1
• Common acute lymphocytic leukemia antigen
• DKFZp686O16152
• EC 3.4.24
• EC 3.4.24.11
• Enkephalinase
• EPN
• Leu-19
• membrane metallo-endopeptidase variant 2
• membrane metallo-endopeptidase
• MGC126681
• MGC126707
• MME
• NEPmembrane metallo-endopeptidase (neutral endopeptidase, enkephalinase, CALLA
• Neprilysin
• Neutral Endopeptidase 24.11
• Neutral endopeptidase
• NKH1
• SFE
• Skin fibroblast elastase

Background

Neprilysin/CD10, also known as NEP and neutral endopeptidase 24.11, is a zinc metallopeptidase expressed at the cell surface of a variety of cells. The enzyme functions both as an endopeptidase with a thermolysin-like specificity and as a dipeptidylcarboxypeptidase. NEP has been shown to be involved in the degradation of enkephalins in the mammalian brain and the inactivation of circulating atrial natriuretic peptide (1, 2). NEP has also been identified as the common acute lymphocytic leukemia antigen (CALLA), and is expressed on the surface of lymphocytes in some disease states (3, 4). These and other observations have resulted in considerable interest in NEP as a target for analgesics and antihypertensive drugs. NEP is also a major degrading enzyme of amyloid beta peptide (A beta ) in the brain, indicating that down-regulation of NEP activity, which could be caused by aging, can contribute to the development of Alzheimer’s disease by promoting A beta accumulation (5).

1. Malfroy, B. et al. (1978) Nature 276:523.
2. Kenny, A.J. and Stephenson, S.L. (1988) FEBS Lett. 232:1.
3. LeTarte, M. et al. (1988) J. Exp. Med. 168:1247.
4. Shipp, M.A. et al. (1988) Proc. Natl. Acad. Sci. USA 85:4819.
5. Itwata, N. et al. (2001) Science 292:1550.

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.

Publications for Neprilysin/CD10 Antibody (MAB11821)(1)

Publication using MAB11821

• Polyhydroxycurcuminoids but not curcumin upregulate neprilysin and can be applied to the prevention of Alzheimer&#039;s disease Sci Rep, 2016-07-13;6(0):29760. 2016-07-13 [PMID: 27407064] (Western Blot, Human)

Bioinformatics

• Gene Symbol: MME
• Uniprot:
  • P08473()