Glu-Plasminogen, CAS No.9001-91-6

CAS: 9001-91-6 Cat. No.: np330965

GRADE & PURITY BioReagent, Native, from Human plasma; Pre-lyophilization Protein Concentration: See COA

Accession #

P00747

Bioactivity

See COA

Descripción general

Purified from fresh frozen human plasma. Plasminogen is activated to the serine protease plasmin via urokinase, streptokinase or tissue plasminogen activator.

The Human glu-plasminogen is homogeneous on SDS-PAGE gels and shows no reduction upon incubation with 2-mercaptoethanol. No Plasmin activity was detected using the chromogenic substrate S-2251.

Extinction Coefficient (1%) = 17.0

The above protein was purified from Human plasma units that test negative for antibodies to HBc, HCV, HIV 1/2, HTLV-I/II, and non reactive for HBsAg, HIV-1 RNA, HCV RNA, HBV DNA and Syphilis by FDA approved tests. Donors are screened for CJD.

Specifications

Product Name

Glu-Plasminogen, CAS No.9001-91-6

Sinónimos

Plasminogen, Glu-Type, Human Plasma | Human Glu-Plasminogen | Glu-Plasminogen from Human plasma | Human glu-Plasminogen Native Protein | PLG | HAE4

Grado

BioReagent, Native

Especificaciones y pureza

BioReagent, Native, from Human plasma; Pre-lyophilization Protein Concentration: See COA

Mecanismos bioquímicos y fisiológicos

Plasmin dissolves the fibrin of blood clots and acts as a proteolytic factor in a variety of other processes including embryonic development, tissue remodeling, tumor invasion, and inflammation. In ovulation, weakens the walls of the Graafian follicle. It

Bioactividad

See COA

Número de adhesión

P00747

Peso molecular previsto

90 kDa

CAS

9001-91-6

Tipo de molécula

Protein

Product Line

Productos biológicos

Almacenamiento y envío

Concentración

from Human plasma; Pre-lyophilization Protein Concentration: See COA

Reconstitución

We suggest hydrating the protein with a good quality water to the original volume (See COA). After hydration aliquot into a useful (one time use) size and freeze at -80℃.

Condiciones de almacenamiento de almacenamiento

Store at 2-8°C

Enviado en

Wet ice

Estabilidad y almacenamiento

Store at 2-8℃ long term. Upon reconstitution, it is recommended to aliquot and freeze at -80℃.

Application Protocols

The following are example research protocols using Glu‑plasminogen (general/literature guidance). Adjust parameters to your assay and instrumentation. Item‑specific concentration, buffer, and activity are Not specified for this item; refer to CoA/Spec Sheet.

1. Chromogenic activation/Activity assay (microplate):

• Prepare buffers: 50 mM Tris‑HCl pH 7.4, 150 mM NaCl. Keep reagents on ice.
• Dispense Glu‑plasminogen into wells at desired concentration; include blanks (buffer only) and inhibitor controls (e.g., aprotinin).
• Initiate with tPA or uPA; in parallel, add chromogenic plasmin substrate (e.g., S‑2251) to all wells.
• Monitor A405 at 25–37°C every 1–2 min for 30–120 min. Derive initial rates and compare conditions (± fibrin fragments, ± lysine analogs).

2. Fibrin clot lysis:

• Form clots in cuvettes or microplates by mixing fibrinogen, thrombin, CaCl2, and Glu‑plasminogen; allow to polymerize.
• Add tPA to initiate lysis; measure turbidity decrease at 340 nm over time. Report half‑lysis time (t50).

3. Casein/Gelatin zymography (plasminogen supplemented gels):

• Cast SDS‑PAGE gels containing substrate and defined plasminogen.
• Run samples (e.g., tPA/uPA), renature, and incubate in assay buffer.
• Stain/destain to visualize clear lytic bands corresponding to activator activity.

Good practices:

• Keep zymogen cold until the start; avoid repeated freeze‑thaw; use low‑bind plastics.
• Include calibration curves and internal controls for inter‑assay comparability.

Biological Roles

Plasminogen (PLG) is the zymogen precursor of plasmin, a trypsin‑like serine protease that drives fibrin clot dissolution and contributes to pericellular proteolysis.

Key roles (literature/general):

• Fibrinolysis: tPA and uPA cleave Glu‑plasminogen to plasmin on fibrin surfaces, yielding targeted proteolysis of fibrin clots. α2‑Antiplasmin and PAI‑1/2 regulate activity.
• Kringle‑mediated targeting: Five kringle domains harbor lysine‑binding sites that support high‑affinity interactions with C‑terminal lysines on fibrin and cell‑surface receptors (e.g., annexin II/S100A10 complex, α‑enolase), localizing activation.
• Matrix remodeling and cell biology: Plasmin activates MMP zymogens and degrades ECM proteins (fibronectin, laminin), facilitating tissue remodeling and cell migration; it can modulate growth factor bioavailability.
• Inflammation and immunity: Plasminogen/plasmin participates in leukocyte trafficking, complement interplay, and pathogen interactions through receptor engagement (literature).
• Conformational regulation: Glu‑plasminogen is predominantly compact in plasma; binding of lysine/analogs or proteolysis (yielding Lys‑plasminogen) favors an open, activation‑competent state.

Biochemical parameters (literature examples):

• Activation is markedly enhanced by fibrin (templating tPA) and by lysine analogs that alter kringle interactions.
• Plasmin exhibits broad substrate specificity with preference for cleavage after Lys/Arg residues in flexible regions; rapid inhibition by α2‑antiplasmin limits diffusion.

This product is supplied for research use only and is not intended for diagnostic or therapeutic use.

Buffer Applications

Buffer composition critically influences Glu‑plasminogen conformation and activation kinetics.

• Basal handling buffers (general):

  • PBS, pH 7.2–7.4; 1× ionic strength (137 mM NaCl, 10 mM phosphate, 2.7 mM KCl) supports native folding and compatibility with most bioassays.
  • Tris‑HCl (20–50 mM, pH 7.4–8.0) + 100–150 mM NaCl for kinetic assays; Tris provides good buffering near neutrality.
  • HEPES (20–50 mM, pH 7.2–7.6) for improved pH stability and low UV background.

• Additives (use case dependent):

  • ε‑Aminocaproic acid (EACA, 1–10 mM) or tranexamic acid to reduce unintended activation by occupying kringle lysine‑binding sites; also useful during storage and sample prep.
  • Aprotinin (10–100 KIU/mL range in literature) to inhibit serine proteases once activation has occurred.
  • Ca2+/Mg2+ are not essential for catalytic activity but may influence cofactor or receptor interactions in cell‑based assays.

• Example recipe (literature, for activation assays): 50 mM Tris‑HCl pH 7.4, 150 mM NaCl, 0.01% Tween‑20 (to reduce adsorption), ± 1–3 mM EACA (pre‑assay), remove EACA just before initiating with tPA/uPA if enhanced activation is desired.

• Electrophoresis/zymography: For plasminogen‑containing gels (casein or gelatin zymography), incorporate defined plasminogen to visualize activator bands; run in Tris‑glycine buffers.

Item‑specific buffer supplied and recommended working buffer: Not specified for this item; refer to CoA/Spec Sheet.

Green Alternatives

Green chemistry considerations are limited for aqueous protein reagents, but experimental design can still minimize environmental impact while preserving protein integrity.

• Reduce hazardous preservatives: If antimicrobial control is required and compatible with your assay, consider low‑toxicity alternatives to sodium azide (e.g., short‑term storage at 2–8°C with sterile technique or single‑use aliquots). Note: compatibility with plasminogen must be verified empirically.
• Minimize single‑use plastics: Use low‑bind, reusable (autoclavable) labware where possible; balance with contamination risk for protease assays.
• Buffer selection: Prefer Good’s buffers (HEPES) with low biological oxygen demand and minimal metal chelation side‑effects over high‑concentration Tris where feasible. Prepare buffers at needed volumes to reduce waste.
• Energy use: Since this item stores at 2–8°C (per Product Data), organize workflows to avoid unnecessary cold‑room time and repeated door openings. For long experiments, maintain samples on ice blocks rather than continuously running chilled recirculators when appropriate.
• Assay formats: Use microplate‑based kinetic assays to reduce reagent volumes compared with cuvette spectrophotometry.

Trade‑offs:

• Eliminating preservatives can increase contamination risk; sterile filtration and aseptic handling may be required.
• Some greener buffer choices may slightly affect plasminogen conformation or activator kinetics; validate equivalency relative to your standard conditions.

Item‑specific formulation or preservative details: Not specified for this item; refer to CoA/Spec Sheet.

Pharmaceutical Uses

This product is for research use only. The following notes describe non‑clinical, laboratory and manufacturing‑adjacent contexts in which Glu‑plasminogen standards are commonly applied (literature/general):

• In‑process controls and QC assays: As a reference substrate/zymogen for validating tissue‑type plasminogen activator (tPA) and urokinase (uPA) potency assays using chromogenic or fibrin clot lysis readouts.
• Diagnostic reagent development: Component in research‑grade fibrinolysis assay kits, calibration curves, and method development for clinical laboratory analyzers (not for diagnostic use unless specifically validated and regulated).
• Device/biomaterial evaluation: Research on surface‑mediated fibrinolysis of blood‑contacting materials under development, assessing plasminogen adsorption/activation.
• Bioprocess research: Control material when characterizing antifibrinolytic agents (e.g., EACA, tranexamic acid) or serpins (PAI‑1, α2‑antiplasmin) interactions during formulation development studies.

Regulatory status and pharmacopoeia:

• No pharmacopeial monograph is implied by this listing; any GMP/clinical use would require separate sourcing and qualification.

Item‑specific certifications, viral screening, and lot release tests: Not specified for this item; refer to CoA/Spec Sheet.

Physical Properties

Item‑specific physicochemical specifications are not provided in the Product Data and should be confirmed on the CoA/Spec Sheet. The following are general/literature descriptors for native human Glu‑plasminogen to aid experimental planning:

• Molecular mass (literature): ~88–92 kDa by SDS‑PAGE (glycoform‑dependent); polypeptide length ~791 aa.
• Isoelectric point (pI, literature): ~6.2–6.5; microheterogeneity due to glycosylation yields multiple pI species.
• Extinction (literature): A280 1.0 ≈ ~1.1–1.3 mg/mL for native Glu‑plasminogen (varies with glycosylation); determine exact factor empirically for your lot.
• Solubility: readily soluble in neutral, buffered aqueous solutions (e.g., PBS, Tris‑HCl, HEPES). Avoid organic solvents and extreme ionic detergents unless denaturation is intended.
• Conformation: compact “closed” Glu‑form at physiological ionic strength; opens in presence of lysine/analogs (e.g., ε‑aminocaproic acid, tranexamic acid) or fibrin surface, which enhances activator binding.
• Stability: susceptible to autoproteolysis upon activation to plasmin; stabilized by serine protease inhibitors (e.g., aprotinin) or lysine analog antifibrinolytics. Avoid repeated freeze‑thaw of solutions.
• Other classical small‑molecule metrics (bp, mp, density, logP, refractive index) are not applicable to proteins.

Notes:

• Any lot‑specific concentration, buffer composition, preservative, and purity metrics are Not specified for this item; refer to CoA/Spec Sheet.
• For accurate concentration, prefer amino acid analysis or UV with a lot‑specific E(1%, 280 nm) if provided.

Quality and Grades

• Item‑specific QC data (purity %, activity, source species, buffer/preservatives): Not specified for this item; refer to CoA/Spec Sheet.

Understanding the provided grade:

• BioReagent, Native (from Product Data): indicates material suitable for general biochemical research applications and isolated from a natural biological source (commonly human plasma for plasminogen). “Native” specifies the protein retains its physiological post‑translational modifications and intact N‑terminal glutamate (Glu‑form), as opposed to recombinant or Lys‑plasminogen.

Implications for experimental use (general):

• Native heterogeneity: glycoforms and minor microheterogeneity may be present; this can slightly affect pI, mass, and binding to lysine/fibrin.
• Functional integrity: native Glu‑plasminogen exhibits the compact conformation and expected activation kinetics with tPA/uPA; confirm lot behavior in your assay context.
• Typical characterization (literature) may include: SDS‑PAGE under reducing/non‑reducing conditions, immunoreactivity to anti‑plasminogen antibodies, mass/IEF profiles, and activity after activation (e.g., chromogenic substrate turnover). These are examples only and are not item‑specific claims.

What to check on the CoA/Spec Sheet:

• Source and fractionation method (e.g., lysine‑Affinity chromatography).
• Buffer composition, pH, ionic strength, and any stabilizers (e.g., ε‑aminocaproic acid, aprotinin) or preservatives.
• Concentration determination method and acceptance criteria.
• Endotoxin/bioburden levels if relevant to your application.

If low UV background or clinical diagnostic grade is required, assess whether this BioReagent, Native grade meets those needs or if a higher specification is warranted.

Reaction and Applications

Glu‑plasminogen is a central reagent for studying fibrinolysis, pericellular proteolysis, and protease activation cascades.

Key applications (literature/general):

• Activation kinetics: Examine conversion of Glu‑plasminogen to plasmin by tPA or uPA in the presence/absence of cofactors (fibrin, plasminogen receptors, or lysine analogs). Monitor plasmin formation via chromogenic substrates (e.g., Spectrozyme PL, S‑2251) or fluorogenic peptides.
• Fibrin clot lysis assays: Incorporate plasminogen into fibrin clots; initiate lysis with tPA and record turbidity decrease or confocal imaging of fibrinolysis fronts.
• Cell‑surface proteolysis: Assess recruitment/activation on cells expressing plasminogen receptors; study invasion/migration in matrix‑coated systems.
• Inhibitor profiling: Evaluate antifibrinolytics (EACA, tranexamic acid), serpins (α2‑antiplasmin, PAI‑1/2), or engineered kringle ligands.
• Proteomics/mapping: After activation, identify plasmin cleavage sites in substrates (fibrin(ogen), ECM proteins) by MS.

Practical notes:

• Maintain zymogen integrity until assay start by cooling, including EACA (1–10 mM) and/or aprotinin.
• Trigger activation with defined amounts of tPA/uPA; include fibrin fragments (e.g., CNBr‑fibrinogen) to accelerate tPA‑mediated activation (literature).
• Control ionic strength and pH (~7.4) to maintain physiological relevance; calcium is not essential for catalytic activity but may affect some cofactor interactions.
• To stop reactions, add serine protease inhibitors (aprotinin, PPACK) or rapidly chill and add SDS sample buffer for SDS‑PAGE analysis.

Item‑specific activity or unit definition: Not specified for this item; refer to CoA/Spec Sheet.

Reaction Conditions

Typical experimental conditions for plasminogen activation and plasmin activity assays (literature/general guidance; optimize for your system):

Activation of Glu‑plasminogen:

• Buffer: 50 mM Tris‑HCl, pH 7.4–7.5; 100–150 mM NaCl; optional 0.01% Tween‑20 to reduce adsorption.
• Cofactors: Fibrin or fibrin fragments (e.g., CNBr‑fibrinogen) markedly accelerate tPA‑mediated activation; lysine analogs modulate conformation and kinetics.
• Temperature: 25–37°C; physiological studies typically at 37°C.
• Initiation: add tPA or uPA at defined molar ratios to plasminogen; monitor plasmin generation continuously using a chromogenic substrate (e.g., p‑nitroanilide‑based) by A405 or fluorogenic peptide by fluorescence.
• Quench: serine protease inhibitors (e.g., aprotinin) or rapid denaturation with SDS sample buffer for endpoint analyses.

Plasmin activity assay (post‑activation):

• Substrate: small‑molecule chromogenic (S‑2251, etc.) or protein substrates (fibrin, casein, gelatin).
• Conditions: neutral pH (7.2–7.6), ionic strength 0.1–0.2 M NaCl; Ca2+ generally dispensable for catalysis.
• Readout: continuous kinetic A405/A405‑derivative or gel zymography (casein/gelatin) with clear zones after incubation.

Notes:

• Maintain zymogen integrity on ice prior to reaction start; remove EACA/tranexamic acid just before initiating if maximal activation is desired.
• Avoid contaminating proteases in buffers/reagents; include carrier protein only if compatible with readouts.

Item‑specific recommended conditions, concentrations, or activity units: Not specified for this item; refer to CoA/Spec Sheet.

Safety and Handling

GHS hazard information for this specific item is not provided in the Product Data. Always consult the SDS for authoritative safety and regulatory details.

• GHS classification, signal word, pictograms, H‑statements: Not specified for this item; refer to SDS.
• General nature: protein biological reagent for laboratory research use only (non‑hazardous under typical use, but handle with standard laboratory precautions). If sourced from human plasma, treat as a potential biohazard per institutional policy, notwithstanding donor screening.

Recommended PPE and practices:

• Wear lab coat, gloves (nitrile), and eye protection when handling.
• Avoid aerosol generation; open vials gently and work in a clean area to minimize contamination and proteolytic activation.
• Prevent skin/eye contact and ingestion; wash hands after use.

Incompatibilities and special risks (general for plasminogen/plasmin systems):

• Avoid contaminating with proteases or activators (e.g., tPA, uPA, trypsin), which can convert plasminogen to active plasmin and cause unwanted proteolysis.
• Strong acids/bases, oxidants, and denaturants (chaotropes, high SDS) can unfold and inactivate the protein.

First‑aid (general):

• Skin/eye contact: rinse with water for several minutes; remove contaminated clothing.
• Inhalation/ingestion: move to fresh air; seek medical attention if symptoms occur.

Spill/cleanup: wipe with absorbent and a suitable laboratory disinfectant (e.g., freshly prepared 0.5–1% sodium hypochlorite), then water. Dispose per local regulations for biological materials.

Solvent Selection

As a soluble plasma glycoprotein, Glu‑plasminogen is handled in aqueous buffers; organic solvent selection is generally not applicable.

• Suitable media (general):

  • Isotonic, neutral buffers such as PBS (pH 7.2–7.4), Tris‑HCl (20–50 mM, pH 7.4–8.0), or HEPES (20–50 mM, pH 7.2–7.6), often with 100–150 mM NaCl.
  • Additives to maintain zymogen state: ε‑aminocaproic acid (EACA, 1–10 mM) or tranexamic acid can stabilize kringle interactions and limit unintended activation by competing for lysine binding sites; aprotinin inhibits serine proteases including plasmin.
  • Protein carriers (if adsorption is a concern): 0.1–1% carrier protein (e.g., HSA) can reduce surface loss, but ensure it does not confound assays.

• Avoid:

  • Significant percentages of organic cosolvents (MeOH, ACN, DMSO) that can denature proteins or modulate kringle‑ligand interactions.
  • Low‑ionic‑strength buffers that may favor conformational opening in the presence of lysine analogs and promote activation.

Comparison (general guidance):

• Tris‑HCl vs PBS: Tris buffers can weakly interfere with some amine‑reactive chemistries; PBS is inert in most protease assays but phosphate can affect certain metal‑dependent systems.
• HEPES: superior pH stability near neutrality and low temperature compared to Tris; low UV absorbance is advantageous for A280 measurements.

Item‑specific buffer composition and solvent constraints: Not specified for this item; refer to CoA/Spec Sheet.

Storage and Reconstitution

• Storage conditions (from Product Data): Store at 2–8°C. Shipped on wet ice. For research use only.

Item‑specific details such as whether the product is supplied lyophilized or in solution, its buffer composition, concentration, stabilizers, and preservatives are Not specified for this item; refer to CoA/Spec Sheet.

General guidance (literature/practice) to maintain Glu‑plasminogen integrity:

• Keep the protein in a neutral buffer (e.g., PBS or Tris‑HCl, pH ~7.4) with physiological ionic strength. Inclusion of ε‑aminocaproic acid (EACA, 1–10 mM) can help preserve the zymogen state during handling; remove EACA before initiating activation assays if desired.
• Minimize proteolysis by maintaining cold chain, using protease‑free consumables, and avoiding contaminating proteases/activators.
• If supplied lyophilized (not specified), reconstitute gently with sterile, chilled buffer. Allow to dissolve without vortexing; mix by gentle inversion. Determine concentration by A280 using an appropriate extinction coefficient or by amino acid analysis.
• Prepare single‑use aliquots to avoid repeated vial openings. Avoid repeated freeze–thaw of any prepared working solutions.

Important: Follow the storage temperature stated for this item (2–8°C). For any alternate storage strategies or long‑term stability claims, consult the lot‑specific CoA/Spec Sheet and SDS.

Structure and Identity

Glu‑Plasminogen is the native, single‑chain zymogen of the broad‑specificity serine protease plasmin (human PLG), bearing an N‑terminal glutamate (Glu1) residue. It is a modular multi‑domain glycoprotein that circulates in plasma.

• SKU: np330965
• Product name: Glu‑Plasminogen (native)
• CAS: 9001-91-6
• Grade/Purity: BioReagent, Native (from Product Data)
• Molecular formula: Not specified for this item; refer to CoA/Spec Sheet.
• Molecular weight: Not specified for this item; refer to CoA/Spec Sheet. (Typical apparent Mr ~88–92 kDa, literature)
• SMILES: Not applicable to large proteins; not provided.
• InChIKey: Not applicable to large proteins; not provided.

Structural features (literature/general):

• Single polypeptide (~791 aa in human PLG) with multiple disulfide bonds and N‑/O‑linked glycans.
• Domain architecture: PAN/apple domain (N‑terminal), followed by five kringle (K1–K5) lysine‑binding modules, and a C‑terminal serine protease domain containing the catalytic triad (His57, Asp102, Ser195 numbering by chymotrypsinogen convention).
• The Glu‑form adopts a more compact “closed” conformation stabilized by intramolecular kringle interactions; lysine/analog binding promotes an “open” conformation conducive to activation.
• Activation: limited proteolysis by tissue‑type or urokinase‑type plasminogen activator (tPA/uPA) generates two‑chain plasmin (heavy + light chains linked by disulfide) with exposed active site.

2D description in words: an elongated, bead‑on‑a‑string protein where kringle domains (looped, disulfide‑rich modules) precede a C‑terminal trypsin‑like protease domain; N‑terminus is glutamate (defining Glu‑plasminogen versus Lys‑plasminogen).

Synthetic Utility

Glu‑plasminogen is a biological macromolecule and is not used as a classical synthetic reagent or building block in organic synthesis. Therefore, conventional synthetic utility (e.g., as a catalyst, coupling partner, or protecting‑group handle) does not apply.

For researchers in chemical biology and proteomics (literature/general):

• As a protease zymogen, it serves as a substrate for activators (tPA/uPA) and, after conversion to plasmin, as a catalyst for peptide bond hydrolysis with preference for Lys/Arg at P1. This enables controlled generation of peptide fragments for mapping plasmin specificity.
• Kringle domain–ligand studies: Native Glu‑plasminogen provides a platform to evaluate binding of lysine analogs, small molecules, or peptidomimetics targeting kringle domains—useful in ligand discovery campaigns.
• Surface conjugation/biomaterials: Immobilized plasminogen can be used to study on‑surface activation and proteolysis relevant to blood‑contacting materials.

If your project requires a chemical reagent for peptide bond manipulation, consider chemical proteases (e.g., CNBr for Met‑specific cleavage) or engineered proteases; Glu‑plasminogen is instead a biological tool for proteolysis research. See Reaction & Applications for practical use cases.

Target Specificity

Not applicable. This listing is a native protein reagent (Glu‑plasminogen), not an affinity reagent such as an antibody or ligand with defined target specificity parameters (antigen/epitope, clone, isotype, species reactivity). For functional interactions of plasminogen with biological partners (e.g., tPA/uPA, fibrin, lysine‑binding receptors), refer to the Biological Roles and Reaction & Applications sections.