TNF RII/TNFRSF1B Protein, Human (HEK293, hFc)

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TNFRII (TNFRSF1B) protein has a high ability to bind with tumor necrosis factor-alpha (TNF-α). TNFRII has pro-apoptotic function. TNFRII recruits TRAF2, induces gene expression and intensively crosstalks with TNF-R1. TNFRII selectively enhances the induction of apoptosis by the death receptor TNFRI. TNFRII Protein, Human (HEK293, hFc) is expressed by HEK293 and has a transmembrane region (L23-D257) with hFc tag at the C-terminus.

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  • Species: Human
  • Source: HEK293
  • Storage:
    Stored at -20°C for 2 years from date of receipt. After reconstitution, it is stable at 4°C for 1 week or -20°C for longer (with carrier protein). It is recommended to freeze aliquots at -20°C or -80°C for extended storage.
  • Biological Activity
  • Technical Parameters
  • Product Properties
  • Documentation
  • References
  • Help & FAQs

Biological Activity

Description

TNFRII (TNFRSF1B) protein has a high ability to bind with tumor necrosis factor-alpha (TNF-α). TNFRII has pro-apoptotic function. TNFRII recruits TRAF2, induces gene expression and intensively crosstalks with TNF-R1. TNFRII selectively enhances the induction of apoptosis by the death receptor TNFRI. TNFRII Protein, Human (HEK293, hFc) is expressed by HEK293 and has a transmembrane region (L23-D257) with hFc tag at the C-terminus[1].

Background

TNFRII (TNFRSF1B) protein is a single-pass type I membrane protein belonging to the tumor necrosis factor (TNF) family. TNFRII is the major signaling receptor for TNF-α. TNFRII protein is highly regulated and typically found in immune system cells[1].
The amino acid sequence of mouse TNFRII protein has low homology between human and rhesus macaque TNFRII protein (less than 85%). The amino acid sequence of TNFRII protein in human and rhesus macaque is very similar (percent identity matrix of 95.88%).
TNFRII induces apoptosis. TNFRII does not directly engage the apoptotic program, but relies on the induction of endogenous, membrane-bound TNF, which subsequently activates TNFRI. TNFRII stimulates the action of the endogenously produced membrane-bound TNF on TNFRI is drastically enhanced. TNFRII competes with TNFRI for the recruitment of newly synthesized TRAF2-bound anti-apoptotic factors, thereby promoting the formation of a caspase-8-activating TNFRI complex. TNFRII competes with TNFRI for binding of TRAF2 and the TRAF2-associated anti-apoptotic cIAP1 and cIAP2 proteins. cIAP1-initiated degradation of TRAF2, which in turn enhances receptor competition for the remaining TRAF2, cIAP1 and cIAP2 molecules. cIAP1 would have an anti-apoptotic function upon recruitment into the TNFRI signalling complex, but would switch to a net proapoptotic function upon recruitment into the TNFRII signalling complex[1][2][3].

Verified Bioactivity

1.Measured by its ability to inhibit the TNF-alpha mediated cytotoxicity in the L-929 mouse fibroblast cells in the presence of the metabolic inhibitor actinomycin D. The ED50 for this effect, in the presence of 0.25 ng/mL of TNF-alpha, is 0.0014 µg/mL corresponding to a specific activity is 7.143×105 units/mg.
2.Measured by its binding ability in a functional ELISA. Immobilized TNF-α at 0.5 μg/mL can bind human TNFR2, the EC50 of human TNFR2 protein is 1-15 ng/mL.

MCE Validation Data

  • Bioactivity - Cell-Based Assay

    Bioactivity - Cell-Based Assay

    Measured by its ability to inhibit the TNF-alpha mediated cytotoxicity in the L-929 mouse fibroblast cells in the presence of the metabolic inhibitor actinomycin D. The ED50 for this effect, in the presence of 0.25 ng/mL of TNF-alpha, is 0.0014 µg/mL corresponding to a specific activity is 7.143×105 units/mg.

Technical Parameters

  • Species Human
  • Source HEK293
  • Tag C-hFc
  • Accession
  • Gene ID
  • Molecular Construction
    • N-term
    • TNFRII (L23-D257)
      Accession # P20333
    • hFc
    • C-term
  • Protein Length

    Extracellular Domain

  • Synonyms

    TNFRSF1B; TNF-R2; Prev. TNFR2; Tumor Necrosis Factor Receptor Superfamily, Member 1B; TNFBR; Tumor Necrosis Factor Receptor Superfamily, Member 1b; P75; Tumor Necrosis Factor Binding Protein 2; TNF-R-II; Tumor Necrosis Factor Beta Receptor; TNF-R75; Solub

  • AA Sequence

    LPAQVAFTPYAPEPGSTCRLREYYDQTAQMCCSKCSPGQHAKVFCTKTSDTVCDSCEDSTYTQLWNWVPECLSCGSRCSSDQVETQACTREQNRICTCRPGWYCALSKQEGCRLCAPLRKCRPGFGVARPGTETSDVVCKPCAPGTFSNTTSSTDICRPHQICNVVAIPGNASMDAVCTSTSPTRSMAPGAVHLPQPVSTRSQHTQPTPEPSTAPSTSFLLPMGPSPPAEGSTGD

  • Predicted Molecular Mass

    54.1 kDa

  • Molecular Weight

    Approximately 66-80 kDa, based on SDS-PAGE under reducing conditions, due to the glycosylation.

  • Glycosylation

    Yes

  • Purity

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

Product Properties

Appearance

Lyophilized powder

Formulation

Lyophilized from a 0.22 μm filtered solution of PBS, 6% trehalose, pH 7.4.

Endotoxin Level

<1 EU/μg, determined by LAL method.

Reconstitution

It is not recommended to reconstitute to a concentration less than 100 μg/mL in ddH2O.

Storage & Stability

Stored at -20°C for 2 years from date of receipt. After reconstitution, it is stable at 4°C for 1 week or -20°C for longer (with carrier protein). It is recommended to freeze aliquots at -20°C or -80°C for extended storage.

Shipping

Room temperature in continental US; may vary elsewhere.

References

Calculators

Reconstitution Calculator

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

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Desired Reconstitution Concentration Desired Reconstitution Concentration
Dilution Calculator

Concentration (start) × Volume (start) = Concentration (final) × Volume (final)

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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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