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Mouse Legumain/Asparaginyl Endopeptidase Antibody

Catalog #: MAB2058
1 Citation Datasheet / COA / SDS
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MAB2058
MAB2058-SP
Detection of Mouse Legumain/Asparaginyl Endopeptidase by Western Blot.
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Mouse Legumain/Asparaginyl Endopeptidase Antibody Summary

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 either lyophilized or as a 0.2 µm filtered solution in PBS.
Label
Unconjugated
Purity
Protein A or G purified from hybridoma culture supernatant

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.

Scientific Data

Western Blot Detection of Mouse Legumain/Asparaginyl Endopeptidase antibody by Western Blot. View Larger

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.

Reconstitution Calculator

Reconstitution Calculator

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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Preparation and Storage

Reconstitution
Reconstitute at 0.5 mg/mL in sterile PBS.
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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

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Citation for Mouse Legumain/Asparaginyl Endopeptidase Antibody

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 Citation: 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., 2009-08-15;69(16):6607-14.
    Species: Mouse
    Sample Types: Cell Lysates
    Applications: Western Blot

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