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Human EGF R (Luc) HEK293 Reporter Cell

For research use only.

  1. Genetically modified cell lines best reflect MOA (Mechanism of Action)
  2. Higher activity and larger assay window for robust and reproducible cell-based bioassay
  3. Comprehensive application data to support assay development and validation
  4. Full tracible record, stringent quality control and validated cell passage stability
  5. Parental cell line legally obtained from internationally recognized cell resource bank and commercially licensed
  6. Global commercial license assistance whenever regulatory filing is required

描述(Description)

The Human EGF R (Luc) HEK293 Reporter Cell was engineered to not only express STAT3 signaling response element, but also express the receptor full length human EGF R (Uniprot: P00533-1). When stimulated with human EGF protein, the EGF/EGF R interaction drives STAT3-mediated luminescence. Inhibition of EGF binding to EGF R by either anti-EGF or anti-EGF R antibodies results in a decrease in luminescence.

应用说明(Application)

• Screen for anti-human EGF R or anti-human EGF neutralizing antibody.

• Screen for human EGF R small molecule inhibitor

EGF R-STAT3 Assay Principles

生长特性(Growth Properties)

Adherent

筛选标记(Selection Marker)

Hygromycin B (40 μg/mL) + Puromycin (2 μg/mL)

培养基(Complete Growth Medium)

DMEM medium + 10% FBS

冻存液(Freeze Medium)

Serum-free cell cryopreservation medium

装量(Quantity)

1 vial contains at least 5×10^6 cells in 1 mL serum-free cryopreservation medium

存储(Storage)

Frozen in liquid nitrogen.

支原体检测(Mycoplasma Testing)

Negative

无菌检测(Sterility Testing)

Negative

使用说明(Instructions for Use)

See data sheet for detailed culturing and assay protocol.

质量管理控制体系(QMS)

  1. 质量管理体系(ISO, GMP)
  2. 质量优势
  3. 质控流程
 

Receptor Assay

EGF R-STAT3 FACS

Expression analysis of human EGF R on Human EGF R (Luc) HEK293 Reporter Cell by FACS.
Cell surface staining was performed on Human EGF R (Luc) HEK293 Reporter Cell or negative control cell using PE-labeled anti-EGF R antibody.

Protocol

 

Application

EGF R-STAT3 APPLICATION

Inhibition of human EGF protein-induced reporter activity by anti-human EGF R neutralizing antibody.
This reporter cell was incubated with serial dilutions of antibodies in the presence of human EGF protein (Cat. No. EGF-H52H3) with a final concentration of 50 ng/mL. The EC50 of anti-human EGF R neutralizing antibody (Cetuximab) is approximately 1.793 μg/mL.

Protocol

EGF R-STAT3 APPLICATION

Inhibition of human EGF protein-induced reporter activity by human EGF R small molecule inhibitor.
This reporter cell was incubated with serial dilutions of inhibitors in the presence of human EGF protein (Cat. No. EGF-H52H3) with a final concentration of 50 ng/mL. The EC50 of human EGF R small molecule inhibitor (Erlotinib) was approximately 0.01 μM.

Protocol

 

Signaling Bioassay

EGF R-STAT3 SIGNALING

Response to human EGF protein (RLU).
The Human EGF R (Luc) HEK293 Reporter Cell was stimulated with serial dilutions of human EGF protein (Cat. No. EGF-H52H3). The EC50 was approximately 56.23 ng/mL.

Protocol

EGF R-STAT3 SIGNALING

Response to human EGF protein (Fold).
The Human EGF R (Luc) HEK293 Reporter Cell was stimulated with serial dilutions of human EGF protein (Cat. No. EGF-H52H3). The max induction fold was approximately 56.

Protocol

 

Passage Stability

EGF R-STAT3 PASSAGE

Passage stability analysis by Signaling Bioassay.
The continuously growing Human EGF R (Luc) HEK293 Reporter Cell was stimulated with serial dilutions of human EGF protein. Human EGF protein stimulated response demonstrates passage stabilization (fold induction and EC50) across passage 10-20.

Protocol

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背景(Background)

The epidermal growth factor receptor (EGFR) belongs to the ErbB family of receptor tyrosine kinases (RTK). The aberrant activity of EGFR has shown to play a key role in the development and growth of tumor cells, where it is involved in numerous cellular responses including proliferation and apoptosis.

Limited Use&License Disclosure

BY USE OF THIS PRODUCT, RESEARCHER AGREES TO BE BOUND BY THE FOLLOWING TERMS OF LIMITED USE OF THIS CELL LINE PRODUCT.

  1. If the researcher is not willing to accept the terms of limited use of this cell line product, and the product is unused, ACRO will accept return of the unused product.
  2. Researchers may use this product for research use only, no commercial use is allowed. "Commercial use" means any and all uses of this product and derivatives by a party for profit or other consideration and may include but is not limited to use in: (1) product manufacture; and (2) to provide a service, information or data; and/or resale of the product or its derivatives, whether or not such product or derivatives are resold for use in research.
  3. This cell line is neither intended for any animal or human therapeutic purposes nor for any direct human in vivo use . You have no right to share, modify, transfer, distribute, sell, sublicense, or otherwise make the cell line available for use to other researchers, laboratories, research institutions, hospitals, universities, or service organizations.
  4. ACROBIOSYSTEMS MAKES NO WARRANTIES OR REPRESENTATIONS OF ANY KIND, EITHER EXPRESSED OR IMPLIED, WITH RESPECT TO THE SUITABILITY OF THE CELL LINE FOR ANY PARTICULAR USE.
  5. ACROBIOSYSTEMS ACCEPTS NO LIABILITY IN CONNECTION WITH THE HANDLING OR USE OF THE CELL LINE.
  6. Modifications of the cell line, transfer to a third party, or commercial use of the cell line may require a separate license and additional fees. Please contact order.cn@acrobiosystems.com for further details.

 

前沿进展

Safety and efficacy of long term asfotase alfa treatment in childhood hypophosphatasia
d'Angelo, Lauriola, Silvestrini et al
Ital J Pediatr (2025) 51 (1), 86
Abstract: Hypophosphatasia (HPP) is a rare inherited disorder characterized by a deficiency of tissue-non-specific alkaline phosphatase (TNSALP) due to loss-of-function variants of the ALPL gene. HPP is characterized by an extremely variable age of onset and clinical presentation, largely depending on the type of genetic disruption. Childhood HPP commonly presents with skeletal deformities, bone fragility, precocious tooth loss, muscle weakness and sometimes neurological implications. Laboratory tests usually document low levels of alkaline phosphatase (ALP), and radiologic investigations show peculiar bone abnormalities. Treatment with human recombinant TNSALP (asfotase alpha, Strensiq®), available since 2015, is associated with a sudden improvement and a good safety profile.A previously healthy 15-month-old girl presented with progressive "genu valgus" and sudden limping. The patient was diagnosed with childhood HPP due to the presence of two ALPL variants, never described in compound heterozygosity: a missense variant c.571G > A, p.(Glu191Lys), and a frameshift deletion c.963delG; p.(Lys322Argfs*44), both classified as pathogenetic. The child was promptly treated with asfotase alpha, and good improvement was quickly obtained. Efficacy, safety, and good tolerance persisted after a long-term follow-up of 6 years.Pediatricians should consider HPP in children presenting with a suggestive clinical phenotype. Calcium-phosphorus metabolism, ALP, and vitamin B6 should always be investigated in suspected cases. Moreover, asfotase alfa represents a safe, well-tolerated, and effective drug in children with HPP.© 2025. The Author(s).
Hindlimb unloading reversibly attenuates osteogenic potential of rat skeletal stem and progenitor cells ex vivo
Markina, Andreeva, Buravkova
Cells Tissues Organs (2025)
Abstract: Prolonged space flights negatively affect skeleton. Stromal cells of mesenchymal origin play a crucial role in maintaining homeostasis and in regulating the physiological remodeling of various tissues, and this has particular significance for bone.Hindlimb unloading (HU) of rats as a ground-based model for simulation of microgravity was implemented. The functional activity of skeletal stem and progenitor cells (SSPCs) from rat femoral bones was assessed in vitro after 2 weeks of HU and after 2 weeks of subsequent recovery of load support (HU+R). To characterize the growth of the SSPCs, the number of population doublings (PD) was calculated. Histochemical detection of the activity of alkaline phosphatase (AP) - an early marker of osteo-differentiation - on day 7, and of extracellular matrix (ECM) mineralization - as a sign of late osteo-differentiation - on day 21, were carried out. Quantitative real-time PCR was performed to detect the expression of the genes encoding proteins associated with the functional activity of osteoprogenitor cells (Pparg, Runx2, Alpl, Cxcl12) and bone tissue homeostasis (Mmp9, Spp1, RANKL, OPG, Ibsp, BMP10, Sost).After HU, a decrease in AP activity and a significant attenuation of extracellular matrix mineralization were detected. There was also significant downregulation of the genes those for bone matrix proteins (RANKL, OPG, Ibsp), and of the master-genes controlling osteo- and adipo-differentiation (Runx2, Alpl), as well as of Mmp9, encoding a regulatory molecule of bone matrix remodeling. By contrast, sclerostin (Sost) was upregulated. After HU+R, the PD, an AP activity and the level of extracellular matrix mineralization were restored.HU leads to inhibition of the osteoplastic function of SSPCs. The presented data are significant for the elucidation of microgravity-induced mechanisms of bone impairment and for the development of countermeasures for astronauts as well as for osteo-deficient patients after prolonged immobilization.S. Karger AG, Basel.
Current Perspectives on Additive Manufacturing and Titanium Surface Nanotopography in Bone Formation
da Costa Valente, Uehara, Lisboa Batalha et al
J Biomed Mater Res B Appl Biomater (2025) 113 (3), e35554
Abstract: This study aimed to assess the impact of manufacturing methods (conventional and additive manufacturing) and surface treatments (polished and nanotopographic) on the physicochemical properties of Ti6Al4V alloy and their correlation with osteoblast cellular behavior. The evaluated groups were Machined Discs (MD), Machined Discs with Treatment (MD-WT), Additive-manufactured Discs (AD), and Additive-manufactured Discs with Treatment (AD-WT). Surface analyses included SEM, AFM, surface roughness, EDS, XRD, surface free energy, and zeta potential. MC3T3-E1 cells were cultured for biological assessments, including cell morphology, viability, gene expression, alkaline phosphatase activity, and mineralization. ANOVA and Holm-Sidak tests were applied (p < 0.05). MD exhibited grooved topography, AD had partially fused spherical particles, while MD-WT and AD-WT showed patterns from chemical treatment (H3PO4 + NaOH). EDS identified additional ions in MD-WT and AD-WT. XRD patterns indicated crystal lattice orientation differences. MD-WT and AD-WT displayed higher surface free energy than MD and AD (p < 0.05). AD had greater roughness (Sa 6.98 μm, p < 0.05). Biological analyses revealed higher cell viability for MD and AD (p < 0.001), higher ALP activity in MD, and lower in AD-WT. Gene expression varied, with MD showing higher Alpl, Ibsp, and Bglap (p < 0.001), and AD-WT showing higher Runx2 (p < 0.001). Mineralized matrix behavior was similar for MD, AD, and MD-WT (p > 0.05). MD and AD surfaces demonstrated superior osteogenic differentiation potential, while AD exhibited greater roughness, lower surface free energy, higher cell viability, and osteoblastic differentiation potential.© 2025 Wiley Periodicals LLC.
Long-Term Outcomes of Early Enzyme Replacement Therapy With Asfotase Alfa in Perinatal Benign Hypophosphatasia: Amelioration of Bone Deformities in a Young Child
Terayama, Kobayashi, Suemitsu et al
Cureus (2025) 17 (2), e78473
Abstract: Hypophosphatasia (HPP) is a congenital skeletal dysplasia. Enzyme replacement therapy (ERT) improves survival and bone mineralization of patients with perinatal severe HPP. However, there are few reports of ERT in patients with perinatal benign HPP, and its long-term efficacy remains unclear. Herein, we report the case of a boy with perinatal benign HPP who was initiated on early ERT with asfotase alpha. The patient was suspected of having HPP because of shortened limbs and deformed long bones on fetal 3D-CT. He was diagnosed with HPP based on clinical manifestations, low serum alkaline phosphatase levels, and ALPL gene variants. At the age of two years and three months, he had muscle weakness and motor developmental delay requiring support to walk, and ERT was initiated. His muscle strength improved immediately, and he could walk independently two months after starting ERT. Shortening and bowed limbs improved, and his body height increased from -3.48 SD (at the age of two years and three months) to -1.71 SD (at the age of nine years and eight months) after starting ERT. Hence, early ERT effectively improves motor development, bone deformity, and short stature in patients with perinatal benign HPP.Copyright © 2025, Terayama et al.
Showing 1-4 of 1229 papers.
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