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SRP0289

Sigma-Aldrich

Cathepsin B Active human

recombinant, expressed in FreeStyle 293-F cells, ≥90% (SDS-PAGE)

Synonym(s):

APP secretase, CPSB

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About This Item

UNSPSC Code:
12352204
NACRES:
NA.54

biological source

human

recombinant

expressed in FreeStyle 293-F cells

Assay

≥90% (SDS-PAGE)

form

aqueous solution

specific activity

≥2228 pmol/min-μg

mol wt

43 kDa

concentration

>1 mg/mL

technique(s)

activity assay: suitable

solubility

water: soluble

suitability

suitable for molecular biology

NCBI accession no.

application(s)

life science and biopharma

shipped in

dry ice

storage temp.

−70°C

Gene Information

human ... CTSB(1508)

General description

Research area: Cell signaling. Human cathepsin B (GenBank Accession No. NM_001908), CD33 signal peptide(amino acids 1-16) + Cathepsin B (amino acids 18-339), with C-terminal HIS tag, MW = 43 kDa, expressed in FreeStyle 293-F cells. Cathepsin B is a cysteine protease, found in the lysosome of normal cells and tissues. Cathepsin B active human is encoded by the gene CTSB, located on human chromosome 8p23.1.

Application

Active human cathepsin B is useful for the study of enzyme kinetics, screening inhibitors, and selectivity profiling. Active human cathepsin B has been used in a study to assess maturation through proenzyme activation and proteolytic processing. Active human cathepsin B has also been used in a study to prepare a catalytically active version from its precursor expressed in Escherichia coli in the form of inclusion bodies.
Cathepsin B Active human has been used as a supplement in serum free media for the treatment of MDA-MB-231 cell line. It has also been used as a component in low pH buffer for collagen II and complement 3 processing.

Biochem/physiol Actions

Cathepsin B Active human primarily is a preproenzyme, glycosylated on rough endoplasmic reticulum. Processing of procathepsin B to a 31 kDa, single chain active form occurs in the lysosome. It is further processed to double chain form with a 25/26 kDa heavy chain and 5 kDa heavy chain in lysosome. Cathepsin B, functions as an exopeptidase at low pH. At higher pH in endosomes or neutral pH, cathepsin B functions as endopeptidase. Cathepsin B expression is upregulated in malignant tumors and facilitates tumor invasion. Cathepsin B might reduce the production of amyloid-β peptide associated with Alzheimerβs disease. It also plays a key role in neuroprotective functions.

Unit Definition

One unit is defined as the amount of enzyme that will cleave 1 pmol of substrate per min at 37°C

Physical form

Formulated in 40 mM Tris-HCl, pH 8.0, 110 mM NaCl, 2.2 mM KCl, 3 mM DTT, 20% glycerol, and 200 mM imidazole.

Preparation Note

Thaw on ice. Upon first thaw, briefly spin tube containing enzyme to recover full content of the tube. Aliquot enzyme into single use aliquots. Store remaining undiluted enzyme in aliquots at -70°C. Note: Enzyme is very sensitive to freeze/thaw cycles.

Legal Information

FreeStyle is a trademark of Invitrogen Corp.

Pictograms

Health hazardExclamation mark

Signal Word

Danger

Hazard Statements

Hazard Classifications

Eye Irrit. 2 - Repr. 1B - Skin Irrit. 2

Storage Class Code

6.1C - Combustible acute toxic Cat.3 / toxic compounds or compounds which causing chronic effects

WGK

WGK 1

Flash Point(F)

Not applicable

Flash Point(C)

Not applicable


Certificates of Analysis (COA)

Search for Certificates of Analysis (COA) by entering the products Lot/Batch Number. Lot and Batch Numbers can be found on a product’s label following the words ‘Lot’ or ‘Batch’.

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The dendritic cell MHC II peptidome derives from a variety of processing pathways and includes peptides with a broad spectrum of HLA-DM sensitivity.
Clement CC, et al.
The Journal of Biological Chemistry, 1863(11), jbc-M115 (2016)
Pericellular cathepsin B and malignant progression.
Roshy S, et al.
Cancer Metastasis Reviews, 22(2-3), 271-286 (2003)
Proteomic analysis of silenced cathepsin B expression suggests non-proteolytic cathepsin B functionality.
Sigloch FC, et al.
Biochimica et Biophysica Acta - Molecular Cell Research, 1863(11), 2700-2709 (2016)
R Kuhelj et al.
European journal of biochemistry, 229(2), 533-539 (1995-04-15)
A cDNA clone encoding human procathepsin B was expressed at a high level in Escherichia coli using a T7 polymerase expression system, resulting in the formation of insoluble cytoplasmic protein aggregates (inclusion bodies). The recombinant product was solubilized and renatured
Antiamyloidogenic and neuroprotective functions of cathepsin B: implications for Alzheimer's disease.
Mueller-Steiner S, et al.
Neuron, 51(6), 703-714 (2006)

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