Invitrogen™ SMAD1 (Total/Phospho) Human InstantOne™ ELISA Kit

The Invitrogen™ SMAD1 (Total/Phospho) Human InstantOne™ ELISA Kit is a fast and reliable assay specifically engineered for the accurate quantification of total and phosphorylated human SMAD1 (Ser463/465) in cell lysates. This kit simplifies the workflow, allowing for sample analysis in approximately one hour, making it ideal for research into growth factor signaling, cellular development, and related pathways.

Principle of the Method

The InstantOne™ ELISA employs a modified sandwich ELISA technique optimized for cell lysates:

1. Simultaneous Binding:
   • All reagents, including the capture and detection antibodies, are added directly to the plate containing the cell lysate.
2. Detection Antibody Binding:
   • The specific antibody binds to both total and phosphorylated forms of SMAD1.
3. Signal Development:
   • A substrate solution (TMB) reacts with the antibody-protein complex, producing a measurable signal proportional to the amount of SMAD1 present.
4. Measurement:
   • The signal intensity is read at 450 nm using a colorimetric microplate reader.

Key Features

1. Dual Measurement:
   • Detects both total and phosphorylated SMAD1 (Ser463/465) in cell lysates.
2. Rapid Protocol:
   • Analyze samples in just 1 hour.
3. Ease of Use:
   • Simplified workflow with all reagents included for efficient analysis.
4. Specificity and Accuracy:
   • Exclusively recognizes human SMAD1, ensuring high precision.

Applications

• Growth Factor Signaling Research:
  • Study SMAD1’s role in TGF-β and BMP signaling pathways.
• Cellular Development Studies:
  • Investigate SMAD1’s involvement in morphogenesis, apoptosis, and cell growth.
• Cancer Research:
  • Explore SMAD1 signaling in tumor progression and metastasis.
• Immune Response Studies:
  • Assess SMAD1’s role in modulating immune pathways.
• Therapeutic Development:
  • Evaluate SMAD1 as a target in drug development for signaling-related disorders.

Target Information

SMAD1:

• Function:
  • Mediates signals from bone morphogenetic proteins (BMPs), regulating transcription and cell signaling.
  • Plays a critical role in apoptosis, cell growth, and immune responses.
• Phosphorylation:
  • Activated by BMP receptor kinase via phosphorylation at Ser463/465.
• Pathway Regulation:
  • Forms complexes with SMAD4 for transcriptional activity.
  • Degraded via ubiquitination mediated by SMURF1 and SMURF2.
• Clinical Relevance:
  • Dysregulation of SMAD1 signaling is associated with developmental disorders, cancer, and immune system diseases.

Kit Specifications

• Target Detection:
  • Total and phosphorylated SMAD1 (Ser463/465).
• Sample Type:
  • Cell lysates.
• Analysis Time:
  • 1 hour.
• Instrument:
  • Colorimetric Microplate Reader.
• Usage:
  • For research use only.

Kit Contents

• Capture Antibody: Specific for SMAD1.
• Detection Antibody Reagent: Binds to total and phosphorylated SMAD1.
• InstantOne™ Assay Plate: Pre-coated plate for simplified workflow.
• Cell Lysis Buffer: Optimized for cell lysate preparation.
• Enhancer Solution: Enhances signal detection.
• Wash Buffer (10X): Ensures thorough washing for high signal-to-noise ratio.
• Substrate Reagent (TMB): Develops the measurable signal.
• Stop Solution: Stops the enzymatic reaction.
• Positive Control Cell Lysate (lyophilized): Validates assay performance.
• Plate Seal: Prevents evaporation and contamination.

Storage and Stability

• Storage Conditions: Store kit components at 2–8°C.
• Shelf Life: Ensure proper storage for long-term reagent stability.

The Invitrogen™ SMAD1 (Total/Phospho) Human InstantOne™ ELISA Kit is a vital tool for researchers studying TGF-β/BMP signaling, cellular development, and immune pathways. Its fast analysis time, dual detection capability, and precise quantification make it indispensable for clinical and experimental research applications.