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PFKM Protein, Human (HEK293, His)

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Based on 1 Customer Validation

PFKM protein, a key regulator in cellular metabolism, catalyzes the phosphorylation of D-fructose 6-phosphate to fructose 1,6-bisphosphate, initiating glycolysis. This fundamental enzymatic step controls glucose entry into glycolysis, influencing downstream energy production and cellular metabolism. PFKM's pivotal role highlights its significance in orchestrating metabolic flux and maintaining cellular energy balance. PFKM Protein, Human (HEK293, His) is the recombinant human-derived PFKM protein, expressed by HEK293 , with C-6*His labeled tag.

For research use only. We do not sell to patients.
  • Species: Human
  • Source: HEK293
  • Storage:
    Stored at -80°C for 1 year from date of receipt. It is stable at -20°C for 3 months after opening. It is recommended to freeze aliquots at -80°C for extended storage. Avoid repeated freeze-thaw cycles.
  • Experimental Validation Results for PFKM Protein, Human (HEK293, His)
  • PFKM Protein, Human (HEK293, His) Product Image
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To place orders, for customer services and technical support, please contact:

609-228-6898
[email protected] [email protected]
Find Your Local Distributor Your local distributor: Hycultec GmbHHycultec
  • Biological Activity
  • Technical Parameters
  • Product Properties
  • Documentation
  • Help & FAQs
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Biological Activity

Description

PFKM protein, a key regulator in cellular metabolism, catalyzes the phosphorylation of D-fructose 6-phosphate to fructose 1,6-bisphosphate, initiating glycolysis. This fundamental enzymatic step controls glucose entry into glycolysis, influencing downstream energy production and cellular metabolism. PFKM's pivotal role highlights its significance in orchestrating metabolic flux and maintaining cellular energy balance. PFKM Protein, Human (HEK293, His) is the recombinant human-derived PFKM protein, expressed by HEK293 , with C-6*His labeled tag.

Background

The PFKM protein assumes a pivotal role in cellular metabolism by catalyzing the phosphorylation of D-fructose 6-phosphate to fructose 1,6-bisphosphate using ATP as a substrate. This enzymatic activity marks the initial and committing step of glycolysis, a fundamental metabolic pathway central to energy production. By facilitating the conversion of fructose 6-phosphate to fructose 1,6-bisphosphate, PFKM plays a key role in regulating the entry of glucose into the glycolytic pathway, thereby influencing downstream energy production and cellular metabolic processes. The significance of PFKM in catalyzing this essential reaction underscores its importance in orchestrating the metabolic flux and maintaining cellular energy balance.

Verified Bioactivity

Measured by its ability to phosphorylate fructose-6-phosphate.The specific activity is >100 pmol/min/μg, as measured under the described conditions.

MCE Validation Data

  • Purity - SDS-PAGE

    Purity - SDS-PAGE

    ≥ 95%, as determined by reducing SDS-PAGE.

Assay Procedure

Materials
Assay buffer (10x): 250 mM HEPES, 1.5 M NaCl, 100 mM MgCl2, 100 mM CaCl2, pH 7.0
PFKM Protein, Human (HEK293, His) (HY-P71090)
Substrate: ATP, Fructose-6-Phosphate (HY-137834)
Standard: ATP

Procedure
1. Dilute the 10x assay buffer with deionized water to prepare 1xassay buffer.
2. Reaction Mixture: Prepare a reaction mixture containing 0.05 mM ATP and 2.5 mM Fructose-6-Phosphate in 1x assay buffer.
3. Dilute Human PFKM to 10 μg/mL in 1x assay buffer.
4. Standard Curve Preparation: Dilute ATP to 0, 0.3, 0.6, 1.2, 2.5, 5, 10, 20, 40, 50 μM in 1x assay buffer. Add 50 μL of each concentration to the wells as standard wells.
5. Experimental Group: Mix 20 μL of 100 μg/mL PFKM protein with 30 μL of reaction mixture.
Blank Group: Mix 20 μL of 1X Assay Buffer with 30 μL of reaction mixture.
6. Add 50 μL of Kinase-LumiTM Enhanced Chemiluminescent Kinase Detection Reagent to Reagent to the standard wells, experimental wells, and blank wells. Mix thoroughly.
7. Incubate at 25oC for 10 minutes, then perform chemiluminescence detection using a multimode microplate reader.
8. Calculate specific activity:

     Specific Activity (pmol/min/μg) =

Adjusted ATP consumption* (nmol) x 1000 (pmol/nmol)
Incubation time (min) x amount of enzyme (μg)

*Adjusted for Substrate Blank
**Derived using calibration standard

Per Well:
Human PFKM: 1.25 μg
ATP: 20 μM
Fructose-6-Phosphate:

Technical Parameters

  • Species Human
  • Source HEK293
  • Tag C-6*His
  • Accession

    P08237-1 (T2-V780)

  • Gene ID
    5213  [NCBI]
  • Molecular Construction
    • N-term
    • PFKM (T2-V780)
      Accession # P08237-1
    • 6*His
    • C-term
  • Protein Length

    Full Length of Isoform-1 Mature Protein

  • Synonyms

    PFKM; Phosphofructokinase Muscle Isoform 4; Prev. PFKX; 6-Phosphofructokinase, Muscle Type; Phosphofructo-1-Kinase Isozyme A; 6-Phosphofructo-1-Kinase; 6-Phosphofructokinase Type A; Phosphofructokinase 1; Phosphohexokinase; Phosphofructokinase-M; PPP1R122

  • AA Sequence

    THEEHHAAKTLGIGKAIAVLTSGGDAQGMNAAVRAVVRVGIFTGARVFFVHEGYQGLVDGGDHIKEATWESVSMMLQLGGTVIGSARCKDFREREGRLRAAYNLVKRGITNLCVIGGDGSLTGADTFRSEWSDLLSDLQKAGKITDEEATKSSYLNIVGLVGSIDNDFCGTDMTIGTDSALHRIMEIVDAITTTAQSHQRTFVLEVMGRHCGYLALVTSLSCGADWVFIPECPPDDDWEEHLCRRLSETRTRGSRLNIIIVAEGAIDKNGKPITSEDIKNLVVKRLGYDTRVTVLGHVQRGGTPSAFDRILGSRMGVEAVMALLEGTPDTPACVVSLSGNQAVRLPLMECVQVTKDVTKAMDEKKFDEALKLRGRSFMNNWEVYKLLAHVRPPVSKSGSHTVAVMNVGAPAAGMNAAVRSTVRIGLIQGNRVLVVHDGFEGLAKGQIEEAGWSYVGGWTGQGGSKLGTKRTLPKKSFEQISANITKFNIQGLVIIGGFEAYTGGLELMEGRKQFDELCIPFVVIPATVSNNVPGSDFSVGADTALNTICTTCDRIKQSAAGTKRRVFIIETMGGYCGYLATMAGLAAGADAAYIFEEPFTIRDLQANVEHLVQKMKTTVKRGLVLRNEKCNENYTTDFIFNLYSEEGKGIFDSRKNVLGHMQQGGSPTPFDRNFATKMGAKAMNWMSGKIKESYRNGRIFANTPDSGCVLGMRKRALVFQPVAELKDQTDFEHRIPKEQWWLKLRPILKILAKYEIDLDTSDHAHLEHITRKRSGEAAV

  • Molecular Weight

    Approximately 93 kDa, based on SDS-PAGE under reducing conditions.

  • Glycosylation

    Yes

  • Purity

    ≥ 95%, as determined by reducing SDS-PAGE.

Product Properties

Appearance

Solution

Formulation

Supplied as a 0.2 μm filtered solution of 20 mM PB, 150 mM NaCl, 5 mM EDTA, 5% Trehalose, pH 6.9.

Endotoxin Level

<1 EU/μg, determined by LAL method.

Storage & Stability

Stored at -80°C for 1 year from date of receipt. It is stable at -20°C for 3 months after opening. It is recommended to freeze aliquots at -80°C for extended storage. Avoid repeated freeze-thaw cycles.

Shipping

Shipping with dry ice.

Calculators

Reconstitution Calculator

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

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Dilution Calculator

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The Specific Activity Calculator Equation
  • Specific Activity (Unit/mg)
  • Biological Activity (ED50)

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

Specific Activity (Unit/mg) Specific Activity (Unit/mg)
Unit/mg
= 106 ÷
Biological Activity (ED50) Biological Activity (ED50)
106 ÷
ng/mL
MOQ
Minimum order quantity
100 mg

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Measured by its ability to inhibit anti-CD3-induced proliferation of stimulated CTLL-2 mouse cytotoxic T cells. The ED50 this effect is 0.8023 μg/ml, corresponding to a specific activity is 1246.42 units/mg.

Assay Types
  • 1/16

    MedChemExpress Validation
     ≥ 95%, as determined by reducing SDS-PAGE.

  • PFKM Protein, Human (HEK293, His) Product Image
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  • Experimental Validation Results for PFKM Protein, Human (HEK293, His)
  • PFKM Protein, Human (HEK293, His) Product Image