Recombinant Mouse Myeloblastin (Prtn3)

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Cat. No.
BT1150705
Source
Yeast
Synonyms
Prtn3Myeloblastin; EC 3.4.21.76; Proteinase 3; PR-3
Purity
>85% (SDS-PAGE)
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Cat. No.
BT1150705
Source
E.coli
Synonyms
Prtn3Myeloblastin; EC 3.4.21.76; Proteinase 3; PR-3
Purity
>85% (SDS-PAGE)
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Cat. No.
BT1150705
Source
E.coli
Synonyms
Prtn3Myeloblastin; EC 3.4.21.76; Proteinase 3; PR-3
Purity
>85% (SDS-PAGE)
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Cat. No.
BT1150705
Source
Baculovirus
Synonyms
Prtn3Myeloblastin; EC 3.4.21.76; Proteinase 3; PR-3
Purity
>85% (SDS-PAGE)
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Cat. No.
BT1150705
Source
Mammalian cell
Synonyms
Prtn3Myeloblastin; EC 3.4.21.76; Proteinase 3; PR-3
Purity
>85% (SDS-PAGE)
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Description

Production and Purification Methods

Recombinant Mouse Prtn3 is generated using mammalian or insect cell systems to mimic native enzymatic activity:

  • Expression: Codon-optimized cDNA cloned into vectors under strong promoters (e.g., CMV or rEF1) .

  • Purification: Affinity chromatography (Ni-NTA for His-tagged proteins) followed by size-exclusion chromatography .

  • Storage: Stable at -80°C in buffers containing 10% glycerol; sensitive to freeze-thaw cycles .

Inflammation and Neutrophil Regulation

  • Proteolytic Activity: Cleaves extracellular matrix proteins (elastin, collagen) and immune mediators like annexin A1, amplifying inflammation .

  • Neutrophil Survival: Transgenic mice expressing human Prtn3 (hPR3Tg) show delayed neutrophil apoptosis and increased peritoneal accumulation during sepsis .

  • Pathological Impact: Sustained Prtn3 activity correlates with tissue damage in models of peritonitis and sepsis .

Myeloid Differentiation and Hematopoiesis

  • HSPC Modulation: Prtn3 deficiency (Prtn3−/− mice) expands hematopoietic stem/progenitor cells (HSPCs) and enhances myeloid differentiation .

  • STAT3 Interaction: Prtn3 binds STAT3, promoting its ubiquitination and degradation, thereby inhibiting myeloid differentiation .

Table 1: Experimental Models and Outcomes

Study ModelKey FindingsSource
Prtn3−/− miceIncreased myeloid progenitors; accelerated recovery post-irradiation .
hPR3Tg miceDelayed inflammation resolution due to annexin A1 cleavage and neutrophil survival .
Competitive BM transplantationPrtn3−/− HSPCs outcompete wild-type cells in repopulation assays .

Leukemia Relevance

  • Prtn3 depletion protects against acute myeloid leukemia (AML) by restoring STAT3-dependent differentiation .

  • Prtn3−/− mice exhibit reduced leukemic burden in xenograft models .

Applications in Research

Recombinant Mouse Prtn3 is widely used for:

ApplicationProtocolReference
In vitro protease assaysMeasures cleavage of substrates like elastin or synthetic peptides .
Neutrophil migration studiesEvaluates endothelial barrier function via PAR-2 activation .
Antibody validationUsed as a standard in ELISA/Western blot for anti-PRTN3 antibodies .

Limitations and Future Directions

  • Species-Specific Differences: Human and mouse Prtn3 exhibit divergent substrate specificities, complicating translational studies .

  • Therapeutic Targeting: Inhibitors of Prtn3 (e.g., serine protease inhibitors) are under exploration for inflammatory diseases and AML .

Product Specs

Form
Lyophilized powder
Note: While we prioritize shipping the format currently in stock, please specify your format preference in order notes for customized preparation.
Lead Time
Delivery times vary depending on purchasing method and location. Contact your local distributor for precise delivery estimates.
Note: All proteins are shipped with standard blue ice packs unless dry ice is specifically requested in advance. Additional fees apply for dry ice shipping.
Notes
Avoid repeated freeze-thaw cycles. Store working aliquots at 4°C for up to one week.
Reconstitution
Centrifuge the vial briefly before opening to consolidate the contents. Reconstitute the protein in sterile, deionized water to a concentration of 0.1-1.0 mg/mL. For long-term storage, we recommend adding 5-50% glycerol (final concentration) and aliquoting at -20°C/-80°C. Our standard glycerol concentration is 50% and can serve as a guideline.
Shelf Life
Shelf life depends on various factors, including storage conditions, buffer composition, temperature, and protein stability. Generally, liquid formulations have a 6-month shelf life at -20°C/-80°C, while lyophilized forms have a 12-month shelf life at -20°C/-80°C.
Storage Condition
Upon receipt, store at -20°C/-80°C. Aliquot for multiple uses to prevent repeated freeze-thaw cycles.
Tag Info
Tag type is determined during the manufacturing process.
The tag type is determined during production. If you require a specific tag, please inform us, and we will prioritize its development.
Synonyms
Prtn3Myeloblastin; EC 3.4.21.76; Proteinase 3; PR-3
Buffer Before Lyophilization
Tris/PBS-based buffer, 6% Trehalose.
Datasheet
Please contact us to get it.
Expression Region
30-250
Protein Length
Full Length of Mature Protein
Purity
>85% (SDS-PAGE)
Species
Mus musculus (Mouse)
Target Names
Prtn3
Target Protein Sequence
I VGGHEARPHS RPYVASLQLS RFPGSHFCGG TLIHPRFVLT AAHCLQDISW QLVTVVLGAH DLLSSEPEQQ KFTISQVFQN NYNPEENLND VLLLQLNRTA SLGKEVAVAS LPQQDQTLSQ GTQCLAMGWG RLGTQAPTPR VLQELNVTVV TFLCREHNVC TLVPRRAAGI CFGDSGGPLI CNGILHGVDS FVIRECASLQ FPDFFARVSM YVDWIQNVLR
Uniprot No.
Q61096

Target Background

Function
Proteinase 3 is a serine protease that degrades elastin, fibronectin, laminin, vitronectin, and collagen types I, III, and IV (in vitro). It enhances endothelial cell barrier function and vascular integrity during neutrophil transendothelial migration by cleaving and activating receptor F2RL1/PAR-2. It may also play a role in neutrophil transendothelial migration, potentially through association with CD177.
Gene References Into Functions
  1. Proteinase 3's involvement in non-alcoholic fatty liver disease and insulin resistance. PMID: 27261776
  2. PR3's role in inducing a microenvironment that promotes pDC-driven Th9/Th2 cell generation in apoptotic cell injection models. PMID: 26436651
  3. PR3-ANCA as a potential biomarker for primary sclerosing cholangitis. PMID: 25397578
  4. Proteinase 3-mediated caspase-3 activation in regulating neutrophil spontaneous death. PMID: 25180606
  5. Increased PRTN3 activity's association with autoimmune diabetes mellitus development in NOD mice. PMID: 25092677
  6. Phagocytosis of apoptotic PR3-expressing cells and its effect on proinflammatory cytokine production in vivo. PMID: 22844112
  7. PR3's dual roles (activation/termination) in IL-33 biological activity. PMID: 22270365
  8. The pathogenic effects of proteinase 3-specific antineutrophil cytoplasmic autoantibodies in inflammation. PMID: 15150076
  9. Differences in interaction patterns between murine and human proteinase 3 complexes and peptidic substrates. PMID: 18023421
  10. PR3 and neutrophil elastase's influence on neutrophil-dependent inflammation through progranulin. PMID: 18568075
  11. Neutrophil-derived PR3's role in proteolyzing high-molecular weight kininogen and activating the kinin pathway. PMID: 19494315
Database Links

KEGG: mmu:19152

STRING: 10090.ENSMUSP00000006679

UniGene: Mm.2364

Protein Families
Peptidase S1 family, Elastase subfamily
Subcellular Location
Lysosome. Secreted. Cell membrane; Peripheral membrane protein; Extracellular side. Membrane raft; Peripheral membrane protein; Extracellular side.

Q&A

FAQs for Recombinant Mouse Myeloblastin (PRTN3) Research

Advanced Research Questions

How does PRTN3 deficiency influence STAT3-dependent myeloid differentiation?

  • Mechanistic approach:

    • Co-immunoprecipitation (Co-IP) assays to confirm PRTN3-STAT3 interaction .

    • Treat Prtn3–/– hematopoietic stem/progenitor cells (HSPCs) with STAT3 inhibitors (e.g., Stattic) to reverse differentiation phenotypes .

    • Track myeloid differentiation via colony-forming assays (methocult GM 3434 medium) .

How to reconcile PRTN3’s proinflammatory role in IC-mediated inflammation with its antileukemic effects?

  • Data contradiction analysis:

    • In inflammation: PRTN3 cleaves anti-inflammatory PGRN, amplifying neutrophil recruitment .

    • In leukemia: PRTN3 inhibits STAT3-driven myeloid differentiation, promoting blast survival .

    • Experimental design: Compare PRTN3 activity in Prtn3–/–Ela2–/– vs. leukemia xenograft models .

What are the technical challenges in detecting PRTN3 in murine models?

  • Solutions:

    • Use antibodies validated against recombinant mouse PRTN3 (e.g., MAB6134) .

    • Pre-treat samples with protease inhibitors to prevent auto-degradation during lysis .

    • Optimize immunofluorescence protocols for azurophilic granule localization .

Functional Assays and Data Interpretation

How to assess PRTN3’s impact on neutrophil survival in vivo?

  • Approach:

    • Compare apoptosis rates in WT vs. Prtn3–/– neutrophils using Annexin V/PI staining .

    • Induce sepsis via cecal ligation and puncture (CLP) and monitor neutrophil accumulation in peritoneal lavage fluid .

What controls are critical for PRTN3-related gene-editing studies?

  • Include Ela2–/– mice to account for NE redundancy .

  • Validate PRTN3 knockout via RT-qPCR (bone marrow cells) and activity assays (casein zymography) .

Key Research Findings

PhenotypeModel SystemKey OutcomeSource
Reduced IC-mediated inflammationPrtn3–/–Ela2–/– mice70% decrease in neutrophil influx vs. WT
Spontaneous myeloid differentiationPrtn3–/– HSPCs2.5-fold increase in GMP populations
Delayed sepsis resolutionhPR3Tg mice100% mortality by Day 5 vs. 50% in WT

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