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Legumain Cell Lysate | LGMN cell lysate

Mouse Legumain HEK293 Overexpression Lysate

Gene Names
Lgmn; AEP; Prsc1; AI746452; AU022324
Applications
Western Blot
Synonyms
Legumain; Mouse Legumain HEK293 Overexpression Lysate; Mouse Legumain/ LGMN HEK293 Cell Lysate (WB positive control); legumain; Mouse AEP Overexpression Lysate; Mouse AI746452 Overexpression Lysate; Mouse AU022324 Overexpression Lysate; Mouse Prsc1 Overexpression Lysate; LGMN cell lysate
Ordering
For Research Use Only!
Host
HEK293 Cells
Form/Format
1x Sample Buffer (1x modified RIPA buffer+1x SDS loading buffer).
Modified RIPA Lysis Buffer: 50 mM Tris-HCl pH 7.4, 150 mM NaCl, 1mM EDTA, 1% Triton X-100, 0.1% SDS, 1% Sodium deoxycholate, 1mM PMSF.
Sequence Positions
434aa
Sequence
Val18-Tyr435
Applicable Applications for LGMN cell lysate
Western Blot (WB)
Application Notes
WB: Use at an assay dependent dilution.
Species
Mouse
Recommend Usage
1. Centrifuge the tube for a few seconds and ensure the pellet at the bottom of the tube.2. Re-dissolve the pellet using 200uL pure water and boil for 2-5 min.
Sequence Construction
A DNA sequence encoding the extracellular domain (Val 18-Tyr 435) of mouse LGMN (NP_035305.1) precursor was expressed with a C-terminal polyhistidine tag.
Preparation Method
Cell lysate was prepared by homogenization of the over-expressed cells in ice-cold modified RIPA Lysis Buffer with cocktail of protease inhibitors. Cell debris was removed by centrifugation. Protein concentration was determined by Bradford assay (protein assay, Microplate Standard assay). The cell lysate was boiled for 5 min in 1 x SDS loading buffer (50mM Tris-HCl pH 6.8, 12.5% glycerol, 1% sodium dodecylsulfate, 0.01% bromophenol blue) containing 5% b-mercaptoethanol, and lyophilized.
Preparation and Storage
Store at 4 degree C for up to twelve months from date of receipt. After re-dissolution, aliquot and store at -80 degree C for up to twelve months. Avoid repeated freeze-thaw cycles.
Samples are stable for up to twelve months from date of receipt.
Related Product Information for LGMN cell lysate
The Mammalian Legumain, also known as LGMN, also called asparaginyl endopeptidase (AEP), is a cysteine protease belonging to peptidase family C13 with strict specificity for hydrolysis of asparaginyl bonds. Known previously only from plants and invertebrates, Legumain is discovered as a lysosomal endopeptidase in mammals. Mammalian Legumain is a cysteine endopeptidase, inhibited by iodoacetamide and maleimides, but unaffected by compound E64. The Mammalian Legumain is involved in the processing of bacterial peptides and endogenous proteins for MHC class II presentation in the lysosomal/endosomal systems. Legumain has been observed to be highly expressed in several types of solid tumors. It was demonstrated in membrane-associated vesicles concentrated at the invadopodia of tumor cells and on cell surfaces where it colocalized with integrins. Legumain was demonstrated to activate progelatinase A. Cells overexpressing Legumain possessed increased migratory and invasive activity in vitro and adopted an invasive and metastatic phenotype in vivo, inferring significance of Legumain in tumor invasion and metastasis. Also, Legumain is expressed in both murine and human atherosclerotic lesions. The macrophage-specific expression of Legumain in vivo and the ability of Legumain to induce chemotaxis of monocytes and endothelial cells in vitro suggest that Legumain may play a functional role in atherogenesis.

This Mouse Legumain overexpression lysate was created in HEK293 Cells and intented for use as a Western blot (WB) positive control. Purification of Legumain protein from the overexpression lysate was verified.
References
Schwarz G, et al. (2002) Characterization of legumain. Biol Chem. 383(11): 1813-6.Liu C, et al. (2003) Overexpression of legumain in tumors is significant for invasion/metastasis and a candidate enzymatic target for prodrug therapy. Cancer Res. 63(11): 2957-64.Murthy RV, et al. (2005) Legumain expression in relation to clinicopathologic and biological variables in colorectal cancer. Clin Cancer Res. 11(6): 2293-9.Gawenda J, et al. (2007) Legumain expression as a prognostic factor in breast cancer patients. Breast Cancer Res Treat. 102(1): 1-6.Clerin V, et al. (2007) Expression of the cysteine protease legumain in vascular lesions and functional implications in atherogenesis. Atherosclerosis. 201(1): 53-66.Lew?"n S, et al. (2008) A Legumain-based minigene vaccine targets the tumor stroma and suppresses breast cancer growth and angiogenesis. Cancer Immunol Immunother. 57(4): 507-15.

NCBI and Uniprot Product Information

NCBI GI #
NCBI GeneID
NCBI Accession #
NCBI GenBank Nucleotide #
UniProt Accession #
Molecular Weight
49,373 Da
NCBI Official Full Name
legumain
NCBI Official Synonym Full Names
legumain
NCBI Official Symbol
Lgmn
NCBI Official Synonym Symbols
AEP; Prsc1; AI746452; AU022324
NCBI Protein Information
legumain
UniProt Protein Name
Legumain
Protein Family
UniProt Gene Name
Lgmn
UniProt Synonym Gene Names
Prsc1
UniProt Entry Name
LGMN_MOUSE

Uniprot Description

LGMN: Has a strict specificity for hydrolysis of asparaginyl bonds. Can also cleave aspartyl bonds slowly, especially under acidic conditions. May be involved in the processing of proteins for MHC class II antigen presentation in the lysosomal/endosomal system. Belongs to the peptidase C13 family.

Protein type: EC 3.4.22.34; Protease

Cellular Component: apical part of cell; late endosome; lysosome

Molecular Function: cysteine-type endopeptidase activity; peptidase activity

Biological Process: negative regulation of multicellular organism growth; negative regulation of neuron apoptosis; proteolysis; proteolysis involved in cellular protein catabolic process; receptor catabolic process; renal system process; response to acidity; vacuolar protein processing

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Product Notes

The LGMN lgmn (Catalog #AAA8116330) is a Cell Lysate produced from HEK293 Cells and is intended for research purposes only. The product is available for immediate purchase. The immunogen sequence is 434aa. AAA Biotech's Legumain can be used in a range of immunoassay formats including, but not limited to, Western Blot (WB). WB: Use at an assay dependent dilution. Researchers should empirically determine the suitability of the LGMN lgmn for an application not listed in the data sheet. Researchers commonly develop new applications and it is an integral, important part of the investigative research process. The amino acid sequence is listed below: Val18-Tyr4 35. It is sometimes possible for the material contained within the vial of "Legumain, Cell Lysate" to become dispersed throughout the inside of the vial, particularly around the seal of said vial, during shipment and storage. We always suggest centrifuging these vials to consolidate all of the liquid away from the lid and to the bottom of the vial prior to opening. Please be advised that certain products may require dry ice for shipping and that, if this is the case, an additional dry ice fee may also be required.

Precautions

All products in the AAA Biotech catalog are strictly for research-use only, and are absolutely not suitable for use in any sort of medical, therapeutic, prophylactic, in-vivo, or diagnostic capacity. By purchasing a product from AAA Biotech, you are explicitly certifying that said products will be properly tested and used in line with industry standard. AAA Biotech and its authorized distribution partners reserve the right to refuse to fulfill any order if we have any indication that a purchaser may be intending to use a product outside of our accepted criteria.

Disclaimer

Though we do strive to guarantee the information represented in this datasheet, AAA Biotech cannot be held responsible for any oversights or imprecisions. AAA Biotech reserves the right to adjust any aspect of this datasheet at any time and without notice. It is the responsibility of the customer to inform AAA Biotech of any product performance issues observed or experienced within 30 days of receipt of said product. To see additional details on this or any of our other policies, please see our Terms & Conditions page.

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