FOSB
(FBJ Murine Osteosarcoma Viral Oncogene Homolog B (FOSB))
Reactivity
Human
Detection Method
Colorimetric
Method Type
DNA-Binding ELISA
Application
ELISA
Purpose
Human fosB Transcription Factor Activity Assay. This assay uses a dsDNA coated plate with canonical fosB binding sequences to semi-quantitatively detect active fosB in lysates or nuclear extracts. Only available in North America.
Sample Type
Cell Lysate, Nuclear Extract
Analytical Method
Semi-Quantitative
Specificity
The olionucleotide/antibody pair provided in this kit recognizes human fosB in whole lysates and nuclear extracts.
Characteristics
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
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
TMB One-Step Substrate Reagent
Stop Solution
Material not included
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
Benchtop rocker or shaker
Microplate reader capable of measuring absorbance at 450 nm
FOSB
Reactivity: Human
Colorimetric
DNA-Binding ELISA
Cell Extracts, Tissue Samples
Application Notes
Optimal working dilution should be determined by the investigator.
Plate
Pre-coated
Protocol
Prepare all reagents and samples as instructed in the manual.
Add 100 μL of sample or positive control to each well.
Incubate 2 h at RT or O/N at 4 °C.
Add 100 μL of prepared primary antibody to each well.
Incubate 1 h at RT.
Add 100 μL of prepared HRP-secondary antibody to each well.
Incubate 1 h at RT.
Add 100 μL of TMB One-Step Substrate Reagent to each well.
Incubate 30 min at RT.
Add 50 μL of Stop Solution to each well.
Read at 450 nm immediately.
Restrictions
For Research Use only
Storage
-20 °C
Storage Comment
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.