Human Legumain/Asparaginyl Endopeptidase Antibody

Catalog # Availability Size / Price Qty
AF2199
AF2199-SP
Detection of Human Legumain/Asparaginyl Endopeptidase by Western Blot.
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Citations (9)
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Human Legumain/Asparaginyl Endopeptidase Antibody Summary

Species Reactivity
Human
Specificity
Detects human Legumain/Asparaginyl Endopeptidase in direct ELISAs and Western blots.
Source
Polyclonal Goat IgG
Purification
Antigen Affinity-purified
Immunogen
Mouse myeloma cell line NS0-derived recombinant human Legumain/Asparaginyl Endopeptidase
Ile18-Tyr433
Accession # Q99538
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
1 µg/mL
See below
Immunoprecipitation
25 µg/mL
Conditioned cell culture medium spiked with Recombinant Human Legumain (Catalog # 2199‑CY), see our available Western blot detection antibodies

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 Example

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

Detection of Human Legumain/
Asparaginyl Endopeptidase by Western Blot.
Western blot shows lysates of human kidney tissue, human heart tissue, and human placenta tissue. PVDF membrane was probed with 1 µg/mL of Goat Anti-Human Legumain/Asparaginyl Endopeptidase Antigen Affinity-purified Polyclonal Antibody (Catalog # AF2199) followed by HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF109). Specific bands were detected for Pro-Legumain/
Asparaginyl Endopeptidase at approximately 56 kDa and mature Legumain/Asparaginyl Endopeptidase 37 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.

Reconstitution Calculator

Reconstitution Calculator

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Preparation and Storage

Reconstitution
Reconstitute at 0.2 mg/mL in sterile PBS.
Reconstitution Buffer Available
Reconstitution Buffer 1 (PBS)
Catalog #
Availability
Size / Price
Qty
RB01
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 whose activity is found in several tissues tested (1, 2). 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 (3). Over-expression of Legumain in tumors is significant for invasion/metastasis (4). 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 the cleavage, which result in the removal of the pro peptides in both C- and N-termini (5). 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 (6‑8). Human Legumain is synthesized as a 433 amino acid precursor with a signal peptide (residues 1‑17). The pro enzyme (residues 18‑433) was expressed with an N-terminal His tag. This activity of Legumain can be inhibited by recombinant human (rh) Cystatin C and rhCystatin E/M and recombinant mouse Cystatin C (R&D Systems, Catalog # 1196‑PI, 1286-PI and 1238-PI, respectively).

References
  1. Chen, J.-M. et al. (1997) J. Biol. Chem. 272:8090.
  2. Tanaka, T. et al. (1996) Cytogenet. Cell Genet. 74:120.
  3. Shirahama-Noda, K. et al. (2003) J. Biol. Chem. 278:33194.
  4. Liu, C. et al. (2003) Cancer Res. 63: 2957.
  5. Li D.N. et al. (2003) J. Biol. Chem. 278:38980.
  6. Chen, J.M. et al. (2001) Biol. Chem. 382:777.
  7. Schwarz, G. et al. (2002) Biol. Chem. 383:1813.
  8. 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

Product Datasheets

Citations for Human 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.

9 Citations: Showing 1 - 9
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  1. Overexpression of asparaginyl endopeptidase is significant for esophageal carcinoma metastasis and predicts poor patient prognosis
    Authors: X Liu, Z Wang, G Zhang, Q Zhu, H Zeng, T Wang, F Gao, Z Qi, J Zhang, R Wang
    Oncol Lett, 2018;15(1):1229-1235.
    Species: Human
    Sample Types: Cell Lysates
    Applications: WB
  2. Suppression of asparaginyl endopeptidase attenuates breast cancer-induced bone pain through inhibition of neurotrophin receptors
    Authors: P Yao, Y Ding, Z Han, Y Mu, T Hong, Y Zhu, H Li
    Mol Pain, 2017;13(0):1.74481E+15.
    Species: Human
    Sample Types: Cell Lysates
    Applications: WB
  3. Glycosylation is important for legumain localization and processing to active forms but not for cystatin E/M inhibitory functions
    Authors: NN Lunde, MH Haugen, KB Bodin Lars, I Damgaard, SJ Pettersen, R Kasem, W Rut, M Drag, M Poreba, HT Johansen, R Solberg
    Biochimie, 2017;0(0):.
    Species: Human
    Sample Types: Cell Lysates
    Applications: WB
  4. The Agaricus blazei-Based Mushroom Extract, Andosan�, Protects against Intestinal Tumorigenesis in the A/J Min/+ Mouse
    PLoS ONE, 2016;11(12):e0167754.
    Species: Mouse
    Sample Types: Whole Tissue
    Applications: IHC - Paraffin embedded
  5. The asparaginyl endopeptidase legumain is essential for functional recovery after spinal cord injury in adult zebrafish.
    Authors: Ma, Liping, Shen, Yan-Qin, Khatri, Harsh P, Schachner, Melitta
    PLoS ONE, 2014;9(4):e95098.
    Species: Zebrafish
    Sample Types: Whole Tissue
    Applications: IHC Fresh
  6. Simvastatin inhibits glucose metabolism and legumain activity in human myotubes.
    Authors: Smith R, Solberg R, Jacobsen L, Voreland A, Rustan A, Thoresen G, Johansen H
    PLoS ONE, 2014;9(1):e85721.
    Species: Human
    Sample Types: Cell Lysates
    Applications: WB
  7. Differential secretome analysis reveals CST6 as a suppressor of breast cancer bone metastasis.
    Cell Res., 2012;22(9):1356-73.
    Species: Human
    Sample Types: Cell Lysates
    Applications: WB
  8. Cystatin E/M suppresses legumain activity and invasion of human melanoma.
    Authors: Briggs JJ, Haugen MH, Johansen HT, Riker AI, Abrahamson M, Fodstad O, Maelandsmo GM, Solberg R
    BMC Cancer, 2010;10(1):17.
    Species: Human
    Sample Types: Cell Lysates
    Applications: WB
  9. Expression of the cysteine protease legumain in vascular lesions and functional implications in atherogenesis.
    Authors: Clerin V, Shih HH, Deng N, Hebert G, Resmini C, Shields KM, Feldman JL, Winkler A, Albert L, Maganti V, Wong A, Paulsen JE, Keith JC, Vlasuk GP, Pittman DD
    Atherosclerosis, 2008;0(0):.
    Species: Human
    Sample Types: Cell Lysates
    Applications: WB

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