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HER2/CD340 Protein, Human (HEK293)

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Based on 1 publication(s) in Google Scholar

The HER2/CD340 protein is a dynamic tyrosine kinase that is essential in the neuregulin receptor complex and regulates microtubule dynamics. Upon activation, it triggers the MEMO1-RHOA-DIAPH1 pathway, inhibits GSK3B and promotes the association of APC and CLASP2 on the cell membrane to achieve microtubule stabilization. HER2/CD340 Protein, Human (HEK293) is the recombinant human-derived HER2/CD340 protein, expressed by HEK293 , with tag free.

For research use only. We do not sell to patients.
  • 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.
  • Experimental Validation Results for HER2/CD340 Protein, Human (HEK293)
  • Experimental Validation Results for HER2/CD340 Protein, Human (HEK293)
  • HER2/CD340 Protein, Human (HEK293) Product Image
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  • Biological Activity
  • Technical Parameters
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Biological Activity

Description

The HER2/CD340 protein is a dynamic tyrosine kinase that is essential in the neuregulin receptor complex and regulates microtubule dynamics. Upon activation, it triggers the MEMO1-RHOA-DIAPH1 pathway, inhibits GSK3B and promotes the association of APC and CLASP2 on the cell membrane to achieve microtubule stabilization. HER2/CD340 Protein, Human (HEK293) is the recombinant human-derived HER2/CD340 protein, expressed by HEK293 , with tag free.

Background

HER2/CD340 Protein, a dynamic protein tyrosine kinase, stands as a pivotal component within diverse cell surface receptor complexes, requiring a coreceptor for efficient ligand binding. Crucially, it plays an indispensable role as part of the neuregulin-receptor complex, with GP30 identified as a potential ligand for this receptor. Beyond its receptor functions, HER2/CD340 Protein intricately regulates the outgrowth and stabilization of peripheral microtubules (MTs). Upon activation, the MEMO1-RHOA-DIAPH1 signaling pathway, initiated by ERBB2 activation, orchestrates the phosphorylation and subsequent inhibition of GSK3B at the cell membrane. This strategic inhibition prevents the phosphorylation of APC and CLASP2, facilitating their association with the cell membrane. Notably, membrane-bound APC enables the localization of MACF1 to the cell membrane, a prerequisite for microtubule capture and stabilization. Within the nucleus, HER2/CD340 Protein is actively involved in transcriptional regulation, associating with the 5'-TCAAATTC-3' sequence in the PTGS2/COX-2 promoter to activate transcription. Furthermore, its engagement in the transcription of rRNA genes by RNA Pol I enhances protein synthesis, contributing to overall cell growth. The multifaceted activities of HER2/CD340 Protein underscore its central role in orchestrating diverse cellular processes, ranging from receptor signaling to microtubule dynamics and transcriptional regulation.

Verified Bioactivity

1.This product does not contain protein kinase domain.
2.Measured by its binding ability in a functional ELISA. Immobilized HER2/CD340 Protein, Human (HEK293) at 2 μg/mL (100 μL/well) can bind herceptin and the EC50 is 6-30 ng/mL.
3.Measured by its ability to block anti-ErbB2 mediated inhibition of BT474 human breast ductal carcinoma cell proliferation and the ED50 is 0.3-2.4 μg/mL in the presence of 0.6 µg/mL Anti-ErbB2/Her2 Monoclonal Antibody.
4.Measured by its ability to block anti-ErbB2 mediated inhibition of SK-BR-3 human breast cancer cell proliferation. The ED50 for this effect is 0.5-0.7 μg/mL in the presence of 0.6 µg/mL Trastuzmab(HY-P9907), corresponding to a specific activity is < 1.8×103 units/mg.
5.Immobilized Human Her2/ERBB2 at 2 μg/ml (100 μl/well) can bind Herceptin, the EC50 of Herceptin is 6.0-30.0 ng/mL.

Assay Procedure

Materials
Cell line used for activity assay: SK-BR-3 cells Culture medium: McCoy’s 5A medium + 10% FBS + 1% P/S HER2/CD340 Protein, Human (HEK293) (HY-P73094)
Penicillin-Streptomycin (100×), Sterile (HY-K1006)
Cell Counting Kit-8 (HY-K0301)
Trastuzmab (HY-P9907)

Procedure
1. Cells were seeded at a density of 4,000 cells per well in a 96-well plate, with a volume of 100 μL per well (the outermost row and column of the 96-well plate were not seeded with cells but filled with an equal volume of PBS solution).
2. Protein was prepared in McCoy's 5A complete medium (containing 0.6 μg/mL Trastuzmab) with 1% FBS, and 100 μL per well was added to 96-well plates so that the final concentration of HER2 protein in each well was 0, 0.0001, 0.001, 0.01, 0.05, 0.1, 0.25, 0.5, 1, 2.5, 5, 10 μg/mL.
3. Cells were incubated at 37 °C in a 5% CO2 incubator for 48 h.
4. After reaching the incubation time, 10 μL of CCK8 solution was added to each well under light-avoidance conditions and incubated at 37 °C, 5% CO2 incubator for 2 h.
5. Using the microplate reader, the OD value at 450 nm was determined.
6. Using GraphPad Prism software, plot a curve with OD values on the y-axis and protein concentration on the x-axis, and calculate the ED50 value.

Technical Parameters

  • Species Human
  • Source HEK293
  • Tag Tag Free
  • Accession

    P04626-1 (T23-T652)

  • Gene ID
    2064  [NCBI]
  • Molecular Construction
    • N-term
    • HER2 (T23-T652)
      Accession # P04626-1
    • C-term
  • Protein Length

    Extracellular Domain

  • Synonyms

    ERBB2; P185erbB2; Prev. NGL; V-Erb-B2 Avian Erythroblastic Leukemia Viral Oncogene Homolog 2 (Neuro/Glioblastoma Derived Oncogene Homolog); HER2; V-Erb-B2 Erythroblastic Leukemia Viral Oncogene Homolog 2, Neuro/Glioblastoma Derived Oncogene Homolog; NEU; V-Erb-B2 Avian Erythroblastic Leukemia Viral Oncoprotein 2 Isoform C; Receptor Tyrosine-Protein Kinase ErbB-2; V-Erb-B2 Avian Erythroblastic Leukemia Viral Oncoprotein 2; Proto-Oncogene C-ErbB-2; Neuroblastoma/Glioblastoma Derived Oncogene Homolog; Proto-Oncogene Neu; Receptor Tyrosine-Protein Kinase ErbB-2 Variant; P185(ErbB2); Truncated HER2 Splice Variant P100; C-ERB-2; Receptor Protein-Tyrosine Kinase; C-ERB2; Metastatic Lymph Node Gene 19; MLN-19; Alternative Protein ERBB2; HER-2; C-Erb B2/Neu Protein; CD340; CD340 Antigen; V-Erb-B2 Avian Erythroblastic Leukemia Viral Oncogene Homolog 2; HER-2/Neu; Tyrosine Kinase-Type Cell Surface Receptor HER2; Herstatin; Neuro/Glioblastoma Derived Oncogene Homolog; VSCN2; Human Epidermal Growth Factor Receptor 2; MLN19; Metastatic Lymph Node Gene 19 Protein; TKR1; Erb-B2 Receptor Tyrosine Kinase 2; MLN 19

  • AA Sequence

    TQVCTGTDMKLRLPASPETHLDMLRHLYQGCQVVQGNLELTYLPTNASLSFLQDIQEVQGYVLIAHNQVRQVPLQRLRIVRGTQLFEDNYALAVLDNGDPLNNTTPVTGASPGGLRELQLRSLTEILKGGVLIQRNPQLCYQDTILWKDIFHKNNQLALTLIDTNRSRACHPCSPMCKGSRCWGESSEDCQSLTRTVCAGGCARCKGPLPTDCCHEQCAAGCTGPKHSDCLACLHFNHSGICELHCPALVTYNTDTFESMPNPEGRYTFGASCVTACPYNYLSTDVGSCTLVCPLHNQEVTAEDGTQRCEKCSKPCARVCYGLGMEHLREVRAVTSANIQEFAGCKKIFGSLAFLPESFDGDPASNTAPLQPEQLQVFETLEEITGYLYISAWPDSLPDLSVFQNLQVIRGRILHNGAYSLTLQGLGISWLGLRSLRELGSGLALIHHNTHLCFVHTVPWDQLFRNPHQALLHTANRPEDECVGEGLACHQLCARGHCWGPGPTQCVNCSQFLRGQECVEECRVLQGLPREYVNARHCLPCHPECQPQNGSVTCFGPEADQCVACAHYKDPPFCVARCPSGVKPDLSYMPIWKFPDEEGACQPCPINCTHSCVDLDDKGCPAEQRASPLT

  • Molecular Weight

    Approximately 90-110 kDa, based on SDS-PAGE under reducing conditions, due to the glycosylation.

  • Glycosylation

    Yes

  • Purity

    ≥ 90%, as determined by reducing SDS-PAGE.

Product Properties

Appearance

Lyophilized powder

Formulation

Lyophilized from a 0.2 μm filtered solution of PBS, pH 7.4, 5% trehalose, 5% mannitol, 0.01% Tween 80.

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.

Calculators

Reconstitution 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
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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
     ≥ 90%, as determined by reducing SDS-PAGE.

  • 1/16

    MedChemExpress Validation
     Measured by its ability to block anti-ErbB2 mediated inhibition of SK-BR-3 human breast cancer cell proliferation. The ED50 for this effect is 0.5701 μg/mL in the presence of 0.6 µg/mL Trastuzmab(HY-P9907).

  • HER2/CD340 Protein, Human (HEK293) Product Image
    1/16
  • Experimental Validation Results for HER2/CD340 Protein, Human (HEK293)
  • Experimental Validation Results for HER2/CD340 Protein, Human (HEK293)
  • HER2/CD340 Protein, Human (HEK293) Product Image