SigPath™ Human/Mouse RB(T821/826) Phosphorylation ELISA Kit

CAT#: ITS-1125-WB11
Product Type: Assay Kit
Species: Human, mouse, rat
Target: Phosphorylated retinoblastoma protein at threonine 821 and threonine 826 residues.
Brand: SigPath
Development Stage: Research-Grade Signal Pathway Assay Development
Short Description
Quantifies target protein phosphorylation levels using an ELISA-based colorimetric method.
Brand
SigPath
Development Stage
Research-Grade Signal Pathway Assay Development
Description
The SigPath™ Human/Mouse RB(T821/826) Phosphorylation ELISA Kit provides a rapid and sensitive solution for quantifying phosphorylated pRB(T821/826) directly in cultured cells. This cell-based system eliminates the need for lysate preparation, utilizing in-cell normalization to total protein to ensure high precision. It is an essential tool for high-throughput screening of cell cycle modulators and drug candidates in cancer and signal transduction research.
Features
Elite Sensitivity & Range: Employs a high-affinity sandwich format to detect endogenous phosphorylated RB with a wide dynamic range and low-nanogram sensitivity.
Dual-Species Versatility: Validated for both human and mouse cell lysates, ensuring seamless cross-model consistency for translational oncology and fundamental research.
Simplified Plate-Based Workflow: Streamlines the quantification of site-specific phosphorylation (T821/826) in a standard 96-well format, significantly reducing processing time.
Rigorous Specificity: Utilizes proprietary antibody pairs that exclusively recognize the T821/826 phospho-epitope without cross-reactivity to total or non-phosphorylated Retinoblastoma protein.
Process Relevance
Monitoring cell cycle progression to support the screening of CDK inhibitors and validate anti-proliferative therapeutic efficacy.
Application Stage
Lead discovery, mechanism of action studies, and preclinical evaluation of cell cycle-targeted drug candidates.
Applications
Quantifying site-specific phosphorylation in cellular assays, screening chemical libraries for kinase inhibition, and analyzing growth factor signaling pathways in human and murine cell lines.
Qualified With
Internal performance validation using reference standards under defined assay conditions.
Target
Phosphorylated retinoblastoma protein at threonine 821 and threonine 826 residues.
Detection Method
Fluorescence Detection (Ex 530 nm / Em 585 nm or Ex 360 nm / Em 457 nm)
Sample Type
Cells
Research Areas
Oncology therapeutics, cell cycle regulation, developmental biology, senescence studies, and regenerative medicine, etc.
Size
100 tests
Data Output Type
Quantitative phosphorylation levels measured via colorimetric endpoint detection (OD 450 nm).
Assay Time
6.5 h
Species
Human, mouse, rat
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
The retinoblastoma protein (RB) is a critical gatekeeper of the G1/S transition. Phosphorylation at T821/826 by CDK/cyclin complexes inactivates its tumor-suppressive capacity, facilitating E2F-mediated transcription and uncontrolled cell proliferation.
Pathways
RB resides at the nexus of the CDK4/6-Cyclin D and p16INK4a signaling axes. Upstream mitogenic signals trigger hyperphosphorylation, releasing E2F transcription factors to drive S-phase entry and cell cycle progression.
For Research Use Only | Not For Clinical Use
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