HER2/CD340 Protein, Human (HEK293, His)
Based on 1 Customer Validation
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, His) is the recombinant human-derived HER2/CD340 protein, expressed by HEK293 , with C-6*His labeled tag.
- Species: Human
- Source: HEK293
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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.
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Biological Activity
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Technical Parameters
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Product Properties
- Documentation
- Help & FAQs
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, His) is the recombinant human-derived HER2/CD340 protein, expressed by HEK293 , with C-6*His labeled tag.
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.Loaded Anti-Human HER2 mAb-Fc on Protein A Biosensor, can bind Human HER2-His with an affinity constant of 1-2 nM as determined in BLI assay.
3.Measured by its ability to block anti-ErbB2 mediated inhibition of SK-BR-3 human breast cancer cell proliferation. The ED50 this effect is ≤1 μg/mL in the presence of 0.6 µg/mL Trastuzmab, corresponding to a specific activity is ≥1×103 units/mg.
4.Trastuzumab (HY-P9907) immobilized on CM5 Chip can bind Human HER2 with an affinity constant of 0.622 nM as determined in a SPR assay.
5.Loaded Anvatabart (HY-P990697) on AHC2 biosensor, can bind HER2/CD340 Protein, Human (HEK293, His) with an affinity constant of 1.171E-09 M as determined in BLI assay.
6.Immobilized Recombinant Human HER2 (C-6His) at 1 μg/mL (100 μL/well) can bind Anti-Human HER2 Antibody (TRA_bio, Research Grade).The ED50 of Anti-Human HER2 Antibody (TRA_bio, Research Grade) is 1.0-10.0 ng/mL.
MCE Validation Data
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Purity - SDS-PAGE
Purity - SDS-PAGE
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Bioactivity - SPR
Bioactivity - SPR
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Bioactivity - BLI
Bioactivity - BLI
Assay Procedure
SK-BR-3 Cell Proliferation Blocking Assay
Materials
Cell line used for assay: SK-BR-3 cells
Cell culture medium: McCoy's 5A medium + 10% FBS + 1% P/S
Target protein: HER2/CD340 protein, human (HEK293, His) (HY-P70254)
Penicillin-streptomycin (100×) (HY-K1006)
Cell Counting Kit-8 (HY-K0301)
Procedure
1. Seed cells into a 96-well plate at a density of 4,000 cells per well, with a volume of 100 μL per well (do not add cells to the outer wells of the 96-well plate; add an equal volume of PBS solution instead).
2. After the cells have adhered, remove the medium from the wells and starve the cells for 12 hours in medium containing 1% FBS.
3. First, resuspend the protein in ddH2O to a concentration of 200 μg/mL. Then, dilute the protein with McCoy’s 5A complete medium containing 1% FBS (and 0.6 μg/mL trastuzumab) to a final concentration of 100 μL per well in the 96-well plate, resulting in final HER2 protein concentrations of 0, 0.0001, 0.001, 0.01, 0.05, 0.1, 0.25, 0.5, 1, 2.5, 5, 50, and 100 μg/mL, respectively.
4. Incubate the cells in a 37°C, 5% CO2 incubator for 48 hours.
5. After the incubation period, add 10 μL of CCK-8 solution to each well under light-protected conditions, and incubate in a 37°C, 5% CO2 incubator for 2 hours.
6. Measure the absorbance (OD) at a wavelength of 450 nm using a microplate reader.
7. 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
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Species Human
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Source HEK293
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Tag C-6*His
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Accession
P04626-1 (T23-T652)
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Protein Length
Extracellular Domain
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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
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AA Sequence
TQVCTGTDMKLRLPASPETHLDMLRHLYQGCQVVQGNLELTYLPTNASLSFLQDIQEVQGYVLIAHNQVRQVPLQRLRIVRGTQLFEDNYALAVLDNGDPLNNTTPVTGASPGGLRELQLRSLTEILKGGVLIQRNPQLCYQDTILWKDIFHKNNQLALTLIDTNRSRACHPCSPMCKGSRCWGESSEDCQSLTRTVCAGGCARCKGPLPTDCCHEQCAAGCTGPKHSDCLACLHFNHSGICELHCPALVTYNTDTFESMPNPEGRYTFGASCVTACPYNYLSTDVGSCTLVCPLHNQEVTAEDGTQRCEKCSKPCARVCYGLGMEHLREVRAVTSANIQEFAGCKKIFGSLAFLPESFDGDPASNTAPLQPEQLQVFETLEEITGYLYISAWPDSLPDLSVFQNLQVIRGRILHNGAYSLTLQGLGISWLGLRSLRELGSGLALIHHNTHLCFVHTVPWDQLFRNPHQALLHTANRPEDECVGEGLACHQLCARGHCWGPGPTQCVNCSQFLRGQECVEECRVLQGLPREYVNARHCLPCHPECQPQNGSVTCFGPEADQCVACAHYKDPPFCVARCPSGVKPDLSYMPIWKFPDEEGACQPCPINCTHSCVDLDDKGCPAEQRASPLT
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Predicted Molecular Mass
70.2 kDa
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Molecular Weight
Approximately 88-120 kDa, band on SDS-PAGE under reducing conditions due to the glycosylation.
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Glycosylation
Yes
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Purity
≥ 95%, as determined by reducing SDS-PAGE.
Product Properties
Lyophilized powder
1.Lyophilized from a 0.22 μm filtered solution of 20 mM PB, 150 mM NaCl, pH 7.4.
2.Lyophilized from a 0.22 μm filtered solution of PBS, pH 7.4.
Please refer to the lot-specific COA for specific buffer information.
<1 EU/μg, determined by LAL method.
It is not recommended to reconstitute to a concentration less than 100 μg/mL in ddH2O. For long term storage it is recommended to add a carrier protein (0.1% BSA, 5% HSA, 10% FBS or 5% Trehalose).
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.
Room temperature in continental US; may vary elsewhere.
Documentation
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Data Sheet (242 KB)
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SDS (252 KB)
- English - EN (252 KB)
- Français - FR (252 KB)
- Deutsch - DE (252 KB)
- Norwegian - NO (252 KB)
- Español - ES (252 KB)
- Swedish - SV (252 KB)
- Italian - IT (252 KB)
- Korean - KR (252 KB)
- Portuguese - PT (252 KB)
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Handling Instructions (2659 KB)
Calculators
Concentration (start) × Volume (start) = Concentration (final) × Volume (final)
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