SigPath™ Human/Mouse 4EBP1(S65) Phosphorylation ELISA Kit

CAT#: ITS-1125-WB6
Product Type: Assay Kit
Species: Human (S65), mouse/rat (S64)
Target: Phosphorylated serine 65 (S65) of the eukaryotic translation initiation factor 4E-binding protein 1 (4EBP1).
Brand: SigPath
Development Stage: Research-Grade Signal Pathway Assay Development
Short Description
Quantifies 4E-BP1 (Ser65) phosphorylation levels in cell lysates using an indirect ELISA method.
Brand
SigPath
Development Stage
Research-Grade Signal Pathway Assay Development
Description
The SigPath™ Human/Mouse 4EBP1(S65) Phosphorylation ELISA Kit provides a rapid and sensitive solution for quantifying p4EBP1(S65) levels in cultured cells. This cell-based system eliminates lysate preparation, featuring in-well total protein normalization to ensure high precision. It is an essential tool for high-throughput screening of inhibitors and activators within the mTOR signaling pathway.
Features
High-Throughput Cell-Based Format: Directly culture and assay cells in 96-well plates, eliminating the need for laborious cell lysate preparation.
Internal Protein Normalization: Total cellular protein is measured in the same well to correct for variations in cell loading and density.
Rapid Diagnostic Workflow: Total assay time is reduced to 6.5 hours, requiring only 2.5 hours of hands-on technical time for completion.
Dual-Species Reactivity: Validated for both human and mouse samples, ensuring high specificity for the S65 phosphorylation site of the 4EBP1 protein.
Process Relevance
Monitoring 4EBP1(S65) phosphorylation levels to validate drug-induced mTOR pathway modulation and assess translation initiation efficiency in cellular models.
Application Stage
Preclinical drug discovery, high-throughput inhibitor screening, and signal transduction research.
Applications
Quantifying p4EBP1(S65) in adherent or suspension cells to evaluate the potency of mTOR/PI3K inhibitors, activators, or siRNA-mediated gene silencing effects during lead optimization.
Qualified With
Internal performance validation using reference standards under defined assay conditions.
Target
Phosphorylated serine 65 (S65) of the eukaryotic translation initiation factor 4E-binding protein 1 (4EBP1).
Detection Method
Fluorescence Detection (Ex 530 nm / Em 585 nm or Ex 360 nm / Em 452 nm)
Sample Type
Cells
Research Areas
Oncology therapeutics, metabolic diseases, pharmacokinetics, protein synthesis regulation, etc.
Size
100 tests
Data Output Type
Fluorometric quantitative measurement of phosphorylation status normalized to total protein content.
Assay Time
6.5 h
Species
Human (S65), mouse/rat (S64)
Storage
Store at −20 °C; upon receipt to maintain reagent stability. Follow recommended handling procedures to ensure consistent assay performance.
Storage Comment
Avoid repeated freeze-thaw cycles. During use, minimize the time components are exposed to room temperature and return them promptly to −20 ℃ storage after handling. Do not store at elevated temperatures for extended periods.
Expiry Date
6 months under recommended storage conditions to ensure optimal performance.
Note
For bulk pricing or custom reagent inquiries, please contact us by email or phone. Products are shipped on ice via FedEx.
Restrictions
For Research Use Only (RUO). Not intended for diagnostic or therapeutic use.
Shipping
Shipped on ice under temperature-controlled conditions.
Background
4EBP1 acts as a critical repressor of cap-dependent translation by sequestering the initiation factor eIF4E. Phosphorylation at Ser65 is the final regulatory "switch" that releases eIF4E, triggering the assembly of the translation machinery.
Pathways
As a primary downstream effector of the mTORC1 complex, 4EBP1 integrates signals from the PI3K/Akt and MAPK pathways. Phosphorylation at S65 signifies active nutrient and growth factor signaling, promoting cell growth and proliferation.
For Research Use Only | Not For Clinical Use
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