Human SOX2 Transcription Factor Activity Assay

RayBio® Human SOX2 Transcription Factor Activity Assay. This assay uses a dsDNA coated plate with canonical SOX2 binding sequences to semi-quantitatively detect active SOX2 in lysates or nuclear extracts. Dry ice shipment (additional fee). 2 - 4 week lead time.

$420.00  
TFEH-SOX2-1
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Assay Format

Specificity:
The olionucleotide/antibody pair provided in this kit recognizes human SOX2 in whole lysates and nuclear extracts.
Number of Targets Detected:
1
Species Detected:
  • Human
Compatible Sample Types:
  • Cell Lysates
  • Nuclear Extracts
Design Principle:
  • Sandwich-based
Method of Detection:
Colorimetric
Quantitative/Semi-Quantitative:
  • Semi-Quantitative
Solid Support:
96-well Microplate

Product Specifications

Size:
1, 2, or 5 x 96-Well Strip Microplate Kit

Protein Information

Accession Number:
  • P48431
Gene ID:
  • 6657
Gene Symbols:
  • SOX2
Protein Name & Synonyms:
Transcription factor SOX-2
Target Species:
  • Human

Introduction

SOX2 is a critical component of the core transcriptional regulatory circuitry that controls pluripotency in ESCs. It is essential for the development of the epiblast in the early mammalian embryo and for the maintenance of embryonic stem cells (ESCs). SOX2 is also necessary for the function and maintenance of neural progenitor cells (NPCs) in the nervous system. Further, SOX2 functions in other adult stem cell and progenitor populations in the gastrointestinal and respiratory tract, as well as in the developing lens, inner ear, taste buds, and testes. Together with OCT4 and NANOG, SOX2 binds to the proximal promoters of large cohort of genes involved in pluripotency and development in later stage. SOX2 is a member of the SOX family of transcription factors, which share a highly conserved high-mobility-group (HMG) DNA binding domain. It interacts with DNA by binding to consensus sequence A/TA/TCAAAG. Although SOX1 and SOX3 have more than 80% sequence similarity with SOX2, SOX2 exerts distinct functions and is indispensable for embryonic development.

Product Features

  • Specific transcription factor-DNA binding assay
  • Perfect alternative to EMSA
  • Easy to perform in an ELISA format
  • Non-radioactive assay
  • High throughput (96-well plate format)
  • Assay can be completed within 5 hours

Application Notes

Kit Components
  • 96-well Strip Microplate pre-coated with DNA probes
  • DNA Binding Buffer
  • Positive Control Sample
  • Specific Competitor DNA probe
  • Non-specific Competitor DNA probe
  • Assay Reagent
  • DTT
  • Wash Buffer
  • Primary Antibody
  • HRP-conjugated Secondary Antibody
  • Antibody Diluent Buffer
  • TMB One-Step Substrate Reagent
  • Stop Solution
Other Materials Required
  • Distilled or deionized water
  • 100 ml and 1 liter graduated cylinders
  • Tubes to prepare sample dilutions
  • Absorbent paper
  • Precision pipettes to deliver 2 µl to 1 ml volumes
  • Adjustable 1-25 ml pipettes for reagent preparation
  • Microplate reader capable of measuring absorbance at 450 nm
Protocol Outline
  1. Prepare all reagents and samples as instructed in the manual.
  2. Add 100 µl of sample or positive control to each well.
  3. Incubate 2 h at RT or O/N at 4 °C.
  4. Add 100 µl of prepared primary antibody to each well.
  5. Incubate 1 h at RT.
  6. Add 100 µl of prepared HRP-secondary antibody to each well.
  7. Incubate 1 h at RT.
  8. Add 100 µl of TMB One-Step Substrate Reagent to each well.
  9. Incubate 30 min at RT.
  10. Add 50 µl of Stop Solution to each well.
  11. Read at 450 nm immediately.

Typical Data

Figure 1
Transcription factor activity assay of SOX2 from nuclear extracts of P19 cells. A. Western-blot result of SOX2 from cytoplasm and nuclear fractions. B. Transcription factor activity assay of SOX2 from nuclear fractions with the RayBio® SOX2 Transcription Factor Activity Assay Kit.

Figure 2
Transcription factor activity assay of SOX2 from nuclear extracts of P19 cells with the specific competitor or non-specific competitor. The result shows specific binding of SOX2 to the conserved DNA binding site.

Storage/Stability

Upon receipt, the positive control should be removed and stored at -20° or -80°C. The remainder of the kit can be stored for up to 6 months at 2-8°C from the date of shipment. Opened Microplate Wells or reagents may be stored for up to 1 month at 2° to 8°C. Return unused wells to the pouch containing desiccant pack, reseal along entire edge.
Note: The kit can be used within one year if the whole kit is stored at -20°C upon receipt. Avoid repeated freeze-thaw cycles.

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Usage

This product is furnished for LABORATORY RESEARCH USE ONLY.
Not for diagnostic or therapeutic use.