Mouse Legumain/Asparaginyl Endopeptidase Antibody

(1 citations)   
  • Species Reactivity
    Mouse
  • Specificity
    Detects mouse Legumain/Asparaginyl Endopeptidase in direct ELISAs and Western blots. In Western blots, approximately 25% cross‑reactivity with recombinant human Legumain is observed.
  • Source
    Monoclonal Rat IgG2A Clone # 301417
  • Purification
    Protein A or G purified from hybridoma culture supernatant
  • Immunogen
    Mouse myeloma cell line NS0-derived recombinant mouse Legumain/Asparaginyl Endopeptidase
    Val18-Tyr435
    Accession # O89017
  • Formulation
    Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied as a 0.2 µm filtered solution in PBS.
  • Label
    Unconjugated
Applications
  •  
    Recommended
    Concentration
    Sample
  • Western Blot
    2 µg/mL
    See below
Please Note: Optimal dilutions should be determined by each laboratory for each application. General Protocols are available in the Technical Information section on our website.
Data Examples
Detection of Mouse Legumain/Asparaginyl Endopeptidase by Western Blot. Western blot shows lysates of J774A.1 mouse reticulum cell sarcoma macrophage cell line. PVDF membrane was probed with 2 µg/mL of Rat Anti-Mouse Legumain/Asparaginyl Endopeptidase Monoclonal Antibody (Catalog # MAB2058) followed by HRP-conjugated Anti-Rat IgG Secondary Antibody (Catalog # HAF005). A specific band was detected for Legumain/Asparaginyl Endopeptidase at approximately 37 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.
Preparation and Storage
  • Reconstitution
    Reconstitute at 0.5 mg/mL in sterile PBS.
  • Shipping
    The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below. *Small pack size (SP) is shipped with polar packs. Upon receipt, store it immediately at -20 to -70 °C
  • 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.
Background: Legumain/Asparaginyl Endopeptidase

Legumain is a lysosomal cysteine protease found in all mouse tissues examined, but was particularly abundant in kidney and placenta (1). Legumain plays a pivotal role in the endosomal/lysosomal degradation system because the Legumain deficiency causes the accumulation of pro cathepsins B, H and L, another group of lysosomal cysteine proteases (2). Over-expression of Legumain in tumors is significant for invasion/metastasis (3). Also known as asparaginyl endopeptidase, it specifically cleaves peptide bonds with Asn at the P1 position. Nevertheless, it also cleaves peptide bonds with Asp at the P1 position. Auto-activation of pro Legumain involves both types of cleavage, which results in the removal of the pro peptides in both C- and N-termini (4). In addition, Legumain activates pro MMP‑2 and processes bacterial antigens for MHC class II presentation and pro thymosin alpha to thymosin alpha 1 and thymosin alpha 11, two acidic peptides with immunoregulatory properties (5‑7). Mouse Legumain is synthesized as a 435 amino acid precursor with a signal peptide (residues 1 to 17). The pro enzyme (residues 18 to 435) was expressed with an N-terminal His tag. The purified pro enzyme can be activated under the conditions as described above. Legumain activity can be inhibited by rmCystatin C and recombinant human cystatins C and E/M (R&D Systems, Catalog # 1238-PI, 1196-PI, and 1286-PI). 

  • References:
    1. Chen, J.M. et al. (1998) Biochem. J. 335:111.
    2. Shirahama-Noda, K. et al. (2003) J. Biol. Chem. 278:33194.
    3. Liu, C. et al. (2003) Cancer Res. 63: 2957.
    4. Li D.N. et al. (2003) J. Biol. Chem. 278:38980.
    5. Chen, J.M. et al. (2001) Biol. Chem. 382:777.
    6. Schwarz, G. et al. (2002) Biol. Chem. 383:1813.
    7. Sarndeses, C.S. et al. (2003) J. Biol. Chem. 278:13286.
  • Entrez Gene IDs:
    5641 (Human); 19141 (Mouse)
  • Alternate Names:
    AEP; Asparaginyl Endopeptidase; cysteine protease 1; Legumain; LGMN; LGMN1; Protease, cysteine 1; protease, cysteine, 1 (legumain); PRSC1EC 3.4.22.34
Related Research Areas
Citations:

R&D Systems personnel manually curate a database that contains references using R&D Systems products. The data collected includes not only links to publications in PubMed, but also provides information about sample types, species, and experimental conditions.

1 Citations: Showing 1 - 1

  1. IFN-producing killer dendritic cells are antigen-presenting cells endowed with T-cell cross-priming capacity.
    Authors: Pletneva M, Fan H, Park JJ, Radojcic V, Jie C, Yu Y, Chan C, Redwood A, Pardoll D, Housseau F
    Cancer Res., 2010;69(16):6607-14.
    Species: Mouse
    Sample Type: Cell Lysates
    Application: WB
Isotype Controls
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Rat IgG2A Isotype Control

Ctrl MAB006 90  
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Normal Rat IgG Control

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Normal Rat IgG Control (Azide Free)

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Rat IgG2A Isotype Control

Ctrl MAB006R  
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Secondary Antibodies
Description Application Cat# Citations Images  

Rat IgG HRP-conjugated Antibody

WB, Simple Western HAF005 7  
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Goat Anti-Rat IgG NorthernLights™ NL557-conjugated Antibody

Flow, IHC, ICC NL013 2
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Goat Anti-Rat F(ab)2 IgG (H+L)
APC-conjugated Antibody

Flow F0113 2  
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Rat F(ab)2 IgG (H+L) PE-conjugated Antibody

Flow F0105B 2  
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Goat Anti-Rat IgG NorthernLights™ NL493-conjugated Antibody

Flow, IHC, ICC NL015 1
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Goat Anti-Rat IgG Biotinylated Antibody

WB BAF005 2
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Rat IgG VisUCyte HRP Polymer Antibody

IHC VC005  
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Goat Anti-Rat IgG Antibody

WB AF005 1
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Rat F(ab)2 IgG (H+L) Fluorescein-conjugated Antibody

Flow F0104B  
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Goat Anti-Rat IgG NorthernLights™ NL637-conjugated Antibody

Flow, IHC, ICC NL014
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Goat Anti-Rat F(ab)2 IgG (H+L) PerCP-conjugated Antibody

Flow F0115 1
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