Legumain/Asparaginyl Endopeptidase Overexpression Lysate

Name: Legumain/Asparaginyl Endopeptidase Overexpression Lysate
Brand: Novus Biologicals
SKU: NBP2-11310
Price: 689 USD
Availability: InStock

Novus Biologicals | Catalog # NBP2-11310

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Product Summary
Product Specifications
Scientific Data
Preparation & Storage
Background
Product Documents
Specific Notices
Research Areas

Key Product Details

Species

Human

Applications

Western Blot

Product Summary for Legumain/Asparaginyl Endopeptidase Overexpression Lysate

Legumain/Asparaginyl Endopeptidase Transient Overexpression Lysate

Expression Host: HEK293T

Plasmid: RC224975

Accession#: NM_005606

Protein Tag: C-MYC/DDK

You will receive 1 vial of lysate (100ug), 1 vial of empty vector negative control (100ug), and 1 vial of 2xSDS sample buffer (250ul). Each vial of cell lysate contains 100ug of total protein (at 1 mg/ml). The 2xSDS Sample Buffer consists of 4% SDS, 125mM Tris-HCl pH6.8, 10% Glycerol, 0.002% Bromophenol blue, 100mM DTT.

Product Specifications

Application Notes

This product is intended for use as a positive control in Western Blot. Overexpression of the target protein was confirmed using an antibody to DDK (FLAG) epitope tag (NBP1-71705) present on the protein construct.

Each vial of cell lysate contains 100ug of total protein which should be sufficient for 20-50 reactions. Depending on over-expression level, antibody affinity and detection system, some lysates can go as low as 0.1 ug per load. We recommend starting with 5ug of cell lysate. Add an equal amount of cell lysate and 2X SDS Sample buffer and boil the SDS samples for 10 minutes before loading.

Theoretical MW

47.6 kDa.
Disclaimer note: The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors.

Type

Overexpression

Scientific Data Images for Legumain/Asparaginyl Endopeptidase Overexpression Lysate

Western Blot: Legumain/Asparaginyl Endopeptidase Overexpression Lysate [NBP2-11310]

Western Blot: Legumain Overexpression Lysate (Adult Normal) [NBP2-11310] Left-Empty vector transfected control cell lysate (HEK293 cell lysate); Right -Over-expression Lysate for Legumain.

Formulation, Preparation, and Storage

Formulation

RIPA buffer

Concentration

The exact concentration of the protein of interest cannot be determined for overexpression lysates. Please contact technical support for more information.

Shipping

The product is shipped with dry ice or equivalent. Upon receipt, store it immediately at the temperature recommended below.

Storage

Store at -80C. Avoid freeze-thaw cycles.

Background: Legumain/Asparaginyl Endopeptidase

Legumain encodes a cysteine protease that has a strict specificity for hydrolysis of asparaginyl bonds. This enzyme may be involved in the processing of bacterial peptides and endogenous proteins for MHC class II presentation in the lysosomal/endosomal systems. Enzyme activation is triggered by acidic pH and appears to be autocatalytic. Protein expression occurs after monocytes differentiate into dendritic cells. A fully mature, active enzyme is produced following lipopolysaccharide expression in mature dendritic cells. Overexpression of this gene may be associated with the majority of solid tumor types. This gene has a pseudogene on chromosome 13. Several alternatively spliced transcript variants have been described, but the biological validity of only two has been determined. These two variants encode the same isoform. [provided by RefSeq]

Alternate Names

AEP, Asparaginyl Endopeptidase, LGMN

Gene Symbol

LGMN

Additional Legumain/Asparaginyl Endopeptidase Products

Product Documents for Legumain/Asparaginyl Endopeptidase Overexpression Lysate

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Certificate of Analysis

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Product Specific Notices for Legumain/Asparaginyl Endopeptidase Overexpression Lysate

HEK293T cells in 10-cm dishes were transiently transfected with a non-lipid polymer transfection reagent specially designed and manufactured for large volume DNA transfection. Transfected cells were cultured for 48hrs before collection. The cells were lysed in modified RIPA buffer (25mM Tris-HCl pH7.6, 150mM NaCl, 1% NP-40, 1mM EDTA, 1xProteinase inhibitor cocktail mix, 1mM PMSF and 1mM Na3VO4, and then centrifuged to clarify the lysate. Protein concentration was measured by BCA protein assay kit.

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Lysates are guaranteed for 6 months from date of receipt.

Related Research Areas

Cysteine Proteases and Regulators

Lysosomal Enzymes

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