Mouse Aminopeptidase N/CD13 Antibody

R&D Systems | Catalog # AF2335

R&D Systems

Key Product Details

Species Reactivity

Validated:

Mouse

Cited:

Human, Mouse, Rat, Transgenic Mouse

Applications

Validated:

Immunohistochemistry, Western Blot, Flow Cytometry, Immunocytochemistry, Immunoprecipitation, CyTOF-ready

Cited:

Immunohistochemistry, Immunohistochemistry-Frozen, Western Blot, Flow Cytometry, Immunocytochemistry, IHC-F, In Situ Hybridization

Label

Unconjugated

Antibody Source

Polyclonal Goat IgG
View all Aminopeptidase N/CD13 Primary Antibodies »

Product Specifications

Immunogen

Mouse myeloma cell line NS0-derived recombinant mouse Aminopeptidase N/CD13
Lys69-Ser966
Accession # P97449

Specificity

Detects mouse Aminopeptidase N/CD13 in direct ELISAs and Western blots.

Clonality

Polyclonal

Host

Goat

Isotype

IgG

Scientific Data Images for Mouse Aminopeptidase N/CD13 Antibody

Aminopeptidase N/CD13 antibody in Mouse Splenocytes by Immunocytochemistry (ICC).

Aminopeptidase N/CD13 in Mouse Splenocytes.

Aminopeptidase N/CD13 was detected in immersion fixed mouse splenocytes using Goat Anti-Mouse Aminopeptidase N/CD13 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF2335) at 15 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Goat IgG Secondary Antibody (red; Catalog # NL001) and counterstained with DAPI (blue). Specific staining was localized to cytoplasm. View our protocol for Fluorescent ICC Staining of Non-adherent Cells.
Aminopeptidase N/CD13 antibody in Mouse Kidney by Immunohistochemistry (IHC-P).

Aminopeptidase N/CD13 in Mouse Kidney.

Aminopeptidase N/CD13 was detected in immersion fixed paraffin-embedded sections of mouse kidney using Goat Anti-Mouse Aminopeptidase N/CD13 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF2335) at 0.1 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 Antigen Retrieval Reagent-Basic (Catalog # CTS013). Tissue was stained using DAB (brown) and counterstained with hematoxylin (blue). Specific staining was localized to apical membrane. View our protocol for IHC Staining with VisUCyte HRP Polymer Detection Reagents.
Detection of Aminopeptidase N/CD13 by Immunocytochemistry/ Immunofluorescence

Detection of Aminopeptidase N/CD13 by Immunocytochemistry/ Immunofluorescence

Glucose uptake in brain pericytes via SGLT2.a Representative example of the expression of SLC5A2 (SGLT2) in cultured human brain microvascular pericytes, HEK-293T, and HK2 cells assessed using RT-PCR and quantification using qPCR (n = 3). The mRNA expression was normalized to that of RNA18S. b Representative immunoblottings for SGLT2, CD13, and beta -actin in cultured pericytes of total lysates or membrane fractions obtained by ultracentrifugation, which were transfected with pcDNA3 or pcDNA3-SLC5A2. Human kidney lysates were used as a positive control for SGLT2 expression. c A representative immunoblotting for SGLT2 in cultured brain pericytes after SLC5A2 overexpression with or without PNGase F treatment. The black arrowhead indicates glycosylated SGLT2 and the white arrowhead indicates unglycosylated SGLT2. d Representative images of immunofluorescence for SGLT2 (green), CD13 (red), and DAPI in a cultured brain pericyte (scale bar = 20 μm). e, f (e) Brain pericytes were incubated in Na+-containing HEPES-buffered saline, Na+-free solution, or Na+-containing solution with 100 or 1,000 nmol/L luseogliflozin for 15 min, followed by incubation with additional 200 μmol/L of 2-NBDG (scale bar = 20 μm). f Fluorescence intensity of 2-NBDG was measured using a microplate reader and normalized to that of Hoechst 33342 (n = 8). Data are presented as dot-plots of individual experiments and mean values ± SD. n.s. (not significant) P > 0.05, ***P < 0.001, ****P < 0.0001 by One-way ANOVA followed by Bonferroni’s post hoc test. SGLT2 sodium-glucose cotransporter 2, Luseo luseogliflozin, pMCAO permanent middle cerebral artery occlusion, 2-NBDG 2-deoxy-2-[(7-nitro-2,1,3-benzoxadiazol-4-yl) amino]-d-glucose. Image collected and cropped by CiteAb from the following open publication (https://www.nature.com/articles/s42003-022-03605-4), licensed under a CC-BY license. Not internally tested by R&D Systems.
Detection of Aminopeptidase N/CD13 by Immunohistochemistry

Detection of Aminopeptidase N/CD13 by Immunohistochemistry

Altered pericyte coverage in mTBI mice. a–b Representative images (a) and quantification (b) showing extravasation of plasma IgG (red) in brain parenchyma 3 days after mTBI. n = 6 mice. Dash line indicates an average value from sham-operated group (n = 5 mice). Scale bar = 50 µm. c Representative images showing CD13-positive pericyte coverage (green), lectin-positive endothelial vascular profiles (blue) and extravasation of fibrin deposits (red) in brain parenchyma of mTBI-affected ipsilateral side or contralateral side 3 days after mTBI. Scale bar = 50 µm. Insert on right: high magnification images of boxed regions in (c1–c3), Scale bar = 10 µm. d–e Quantification of CD13-positive pericyte cell bodies per mm2 (d) and pericyte coverage on lectin microvessel profiles (e) in brain parenchyma of mTBI-affected ipsilateral side and contralateral side 3 days after mTBI. n = 9 mice per group. f Pearson’s correlation plot between BBB impairment based on fibrin deposits and loss of pericyte coverage in brain parenchyma 3 days after mTBI. n = 18 hemispheres. r = Pearson's coefficient. P, significance. g Quantification of microvascular density in the cortexes of mTBI-affected ipsilateral side and contralateral side 3 days after mTBI. n = 9 mice per group. Mean ± SD in b, d–e, g; **, P < 0.01; ***, P < 0.001; NS, non-significant (P > 0.05) by Student’s t-test Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/33892818), licensed under a CC-BY license. Not internally tested by R&D Systems.
Detection of Aminopeptidase N/CD13 by Immunohistochemistry

Detection of Aminopeptidase N/CD13 by Immunohistochemistry

Astroglia derived from NPC grafted into CNS lesions exhibit features of border-forming astroglia.a Detailed IHC images showing the astrocyte border phenotype of stroke lesions at 4 weeks in untreated stroke only and NPC grafted strokes. b Survey IHC of NPC grafted SCI lesions at 2 weeks showing NPC-derived astrocyte borders formed around Cd13-positive inflammatory cells. c Detailed IHC images showing the astrocyte border phenotype in SCI derived from transplanted NPC and not from host astrocytes (TK-positive). d, e Survey and detailed, higher magnification 3D IHC imaging of healthy mouse cortex and meninges showing higher expression of transcriptional regulator Id3 (inhibitor of DNA binding 3) in host astrocytes (HA+ Gfap-Cre-RiboTag) that form glia limitans at the meninges and penetrating blood vessels. f High magnification 3D IHC images showing astrocyte borders formed at stroke lesions upregulate expression of Id3. g High magnification 3D IHC images showing grafted NPC that remodel stroke lesions also express Id3. h NPC grafted into SCI lesions that form astrocyte borders also express Id3. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/36171203), licensed under a CC-BY license. Not internally tested by R&D Systems.

Applications for Mouse Aminopeptidase N/CD13 Antibody

Application
Recommended Usage

CyTOF-ready

Ready to be labeled using established conjugation methods. No BSA or other carrier proteins that could interfere with conjugation.

Flow Cytometry

2.5 µg/106 cells
Sample: bEnd.3 mouse endothelioma cell line

Immunocytochemistry

5-15 µg/mL
Sample: Immersion fixed mouse splenocytes

Immunohistochemistry

0.1-15 µg/mL
Sample: Immersion fixed paraffin-embedded sections of mouse kidney

Immunoprecipitation

25 µg/mL
Sample: Conditioned cell culture medium spiked with Recombinant Mouse Aminopeptidase N/CD13 (Catalog # 2335-ZN), see our available Western blot detection antibodies

Western Blot

0.1 µg/mL
Sample: Recombinant Mouse Aminopeptidase N/CD13 (Catalog # 2335-ZN)

Reviewed Applications

Read 1 review rated 5 using AF2335 in the following applications:

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Advanced Features

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Flow Cytometry

Formulation, Preparation, and Storage

Purification

Antigen Affinity-purified

Reconstitution

Reconstitute at 0.2 mg/mL in sterile PBS. For liquid material, refer to CoA for concentration.

Formulation

Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied either lyophilized or as a 0.2 µm filtered solution in PBS.

Shipping

Lyophilized product is shipped at ambient temperature. Liquid small pack size (-SP) is shipped with polar packs. Upon receipt, store 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 reconstitution.
  • 6 months, -20 to -70 °C under sterile conditions after reconstitution.

Calculators

The reconstitution calculator allows you to quickly calculate the volume of a reagent to reconstitute your vial. Simply enter the mass of reagent and the target concentration and the calculator will determine the rest.

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Background: Aminopeptidase N/CD13

The mouse Anpep gene encodes Aminopeptidase N (APN), which is also known as microsomal aminopeptiase, alanyl aminopeptidase, Aminopeptidase M, CD13, or membrane protein p161 (1‑3). The deduced amino acid sequence of mouse APN consists of a short cytoplasmic tail (residues 2 to 8), a transmembrane region (residue 9 to 32), a Ser/Thr rich region and a zinc metalloprotease domain (residues 69 to 966). Widely expressed in many cells, tissues and species, APN cleaves the N-terminal amino acids from bioactive peptides, leading to their inactivation or degradation. The roles of APN in many fields, such as neuroscience, hematopoeitic cells, immune system, angiogenesis, cancer and viral infection, have been reviewed (4).

References

  1. Chen, H. et al. (1996) J. Immunol. 157:2593.
  2. Larsen, S.L. et al. (1996) J. Exp. Med. 184:183.
  3. Hansen, A.S. et al. (1993) Eur. J. Immunol. 23:2358.
  4. Turner, A.J. (2004) in Handbook of Proteolytic Enzymes (ed. Barrett, et al.) p. 289 Academic Press, San Diego.

Alternate Names

Aminopeptidase M, ANPEP, APN, CD13, gp150, PEPN

Entrez Gene IDs

290 (Human); 16790 (Mouse); 397520 (Porcine); 102115663 (Cynomolgus Monkey)

Gene Symbol

ANPEP

UniProt

P97449

Additional Aminopeptidase N/CD13 Products

Product Documents for Mouse Aminopeptidase N/CD13 Antibody

Certificate of Analysis

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Product Specific Notices for Mouse Aminopeptidase N/CD13 Antibody

For research use only

Citations for Mouse Aminopeptidase N/CD13 Antibody

Customer Reviews for Mouse Aminopeptidase N/CD13 Antibody (1)

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Customer Images

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  • Mouse Aminopeptidase N/CD13 Antibody
    Name: Anonymous
    Application: Immunohistochemistry-Frozen
    Sample Tested: Mouse brain
    Species: Mouse
    Verified Customer | Posted 07/25/2022
    Brain pericytes surrounding blood vessels were stained with Mouse Aminopeptidase N/CD13 antibody.
    Mouse brain tissue (12 micrometer thickness) was incubated overnight with AF2335, 1:500, in PBS 1x at 4C. The day after, the slide was washed 3 x PBS 1x and then incubated with secondary anti-goat antibody AlexaFluor 568-conjugated, 1:1000 for 2 hours at r.t. After counterstaining with DAPI, image was acquired with confocal microscopy. AF2335 signal identifies brain pericytes surrounding blood vessels.
    Mouse Aminopeptidase N/CD13 Antibody AF2335

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