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  4. Legumain Protein, Mouse (HEK293, C-His)

Legumain Protein, Mouse (HEK293, C-His)

Cat. No.: HY-P70216A
COA Handling Instructions

Legumain proteins can slowly cleave aspartyl bonds, especially under acidic conditions. Legumain Protein, Mouse (HEK293, C-His) is the recombinant mouse-derived Legumain protein, expressed by HEK293 , with C-10*His labeled tag. The total length of Legumain Protein, Mouse (HEK293, C-His) is 418 a.a., with molecular weight of ~60.0 kDa.

For research use only. We do not sell to patients.

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  • Biological Activity

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Description

Legumain proteins can slowly cleave aspartyl bonds, especially under acidic conditions. Legumain Protein, Mouse (HEK293, C-His) is the recombinant mouse-derived Legumain protein, expressed by HEK293 , with C-10*His labeled tag. The total length of Legumain Protein, Mouse (HEK293, C-His) is 418 a.a., with molecular weight of ~60.0 kDa.

Background

Legumain protein exhibits a strict specificity for the hydrolysis of asparaginyl bonds. Additionally, it demonstrates the ability to cleave aspartyl bonds slowly, particularly in acidic conditions, further expanding its enzymatic versatility. Functionally, Legumain is integral to the processing of proteins for MHC class II antigen presentation within the lysosomal/endosomal system. It also plays a crucial role in MHC class I antigen presentation in cross-presenting dendritic cells by facilitating the cleavage and maturation of Perforin-2 (MPEG1), thereby promoting antigen translocation in the cytosol, as indicated by recent research findings. Moreover, Legumain is essential for normal lysosomal protein degradation in renal proximal tubules and is required for the degradation of internalized EGFR, highlighting its importance in cellular processes and the regulation of cell proliferation.

Biological Activity

1. Measured in a cell proliferation assay using RAW264.7 cells. The ED50 this effect is 0.07438ng/mL, corresponding to a specific activity is 1.344×107 units/mg.
2. Measured by its ability to cleave the fluorogenic peptide substrate, N-carbobenzyloxy-Ala-Ala-Asn-7-amido-4-methylcoumarin (Z-AAN-AMC). The specific activity is >130 pmol/min/µg, as measured under the described conditions.

  • Measured in a cell proliferation assay using RAW264.7 cells.The ED50 for this effect is 0.07438 ng/mL, corresponding to a specific activity is 13444474.3211 units/mg.
Species

Mouse

Source

HEK293

Tag

C-His

Accession

O89017 (V18-Y435)

Gene ID

19141  [NCBI]

Molecular Construction
N-term
Legumain (V18-Y435)
Accession # O89017
10*His
C-term
Synonyms
rMuLegumain/Asparaginyl Endopeptidase, His; Legumain; Lgmn; Asparaginyl endopeptidase; Protease cysteine 1; Prsc1
AA Sequence

VPVGVDDPEDGGKHWVVIVAGSNGWYNYRHQADACHAYQIIHRNGIPDEQIIVMMYDDIANSEENPTPGVVINRPNGTDVYKGVLKDYTGEDVTPENFLAVLRGDAEAVKGKGSGKVLKSGPRDHVFIYFTDHGATGILVFPNDDLHVKDLNKTIRYMYEHKMYQKMVFYIEACESGSMMNHLPDDINVYATTAANPKESSYACYYDEERGTYLGDWYSVNWMEDSDVEDLTKETLHKQYHLVKSHTNTSHVMQYGNKSISTMKVMQFQGMKHRASSPISLPPVTHLDLTPSPDVPLTILKRKLLRTNDVKESQNLIGQIQQFLDARHVIEKSVHKIVSLLAGFGETAERHLSERTMLTAHDCYQEAVTHFRTHCFNWHSVTYEHALRYLYVLANLCEAPYPIDRIEMAMDKVCLSHY

Molecular Weight

Approximately 55-60 kDa

Purity
  • Greater than 95% as determined by reducing SDS-PAGE.
Appearance

Lyophilized powder.

Formulation

Lyophilized from a 0.2 μm filtered solution of 50 mM Tris-HCL/Tris, 300 mM NaCl, pH 7.4.

Endotoxin Level

<1 EU/μg, determined by LAL method.

Reconstitution

It is not recommended to reconstitute to a concentration less than 100 μg/mL in ddH2O. For long term storage it is recommended to add a carrier protein (0.1% BSA, 5% HSA, 10% FBS or 5% Trehalose).

Storage & Stability

Stored at -20°C for 2 years. After reconstitution, it is stable at 4°C for 1 week or -20°C for longer (with carrier protein). It is recommended to freeze aliquots at -20°C or -80°C for extended storage.

Shipping

Room temperature in continental US; may vary elsewhere.

Documentation

Legumain Protein, Mouse (HEK293, C-His) Related Classifications

Help & FAQs
  • Do most proteins show cross-species activity?

    Species cross-reactivity must be investigated individually for each product. Many human cytokines will produce a nice response in mouse cell lines, and many mouse proteins will show activity on human cells. Other proteins may have a lower specific activity when used in the opposite species.

  • Reconstitution Calculator

  • Dilution Calculator

  • Specific Activity Calculator

The reconstitution calculator equation

Volume (to add to vial) = Mass (in vial) ÷ Desired Reconstitution Concentration

Volume (to add to vial) = Mass (in vial) ÷ Desired Reconstitution Concentration
= ÷

The dilution calculator equation

Concentration (start) × Volume (start) = Concentration (final) × Volume (final)

This equation is commonly abbreviated as: C1V1 = C2V2

Concentration (start) × Volume (start) = Concentration (final) × Volume (final)
× = ×
C1   V1   C2   V2

The specific activity calculator equation

Specific Activity (Unit/mg) = 106 ÷ Biological Activity (ED50)

Specific Activity (Unit/mg) = 106 ÷ Biological Activity (ED50)
Unit/mg = 106 ÷ ng/mL

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Legumain Protein, Mouse (HEK293, C-His)
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HY-P70216A
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