Protein Name & Synonyms:
Myc proto-oncogene protein, Class E basic helix-loop-helix protein 39 (bHLHe39), Proto-oncogene c-Myc, Transcription factor p64
- HER/ErbB Signaling
- IGF Signaling
- JAK/STAT Signaling
- MAPK Signaling
- Notch Signaling
- Wnt/beta-Catenin Signaling
The olionucleotide/antibody pair provided in this kit recognizes human TFEB in whole lysates and nuclear extracts.
Number of Targets Detected:
Compatible Sample Types:
- Cell Lysates
- Nuclear Extracts
Method of Detection:
1, 2, or 5 x 96-Well Strip Microplate Kit
c-Myc is a transcription factor that belongs to the Myc family of proteins consisting of c-, N-, and L-Myc. Myc proteins are members of the basic helix-loop-helix (bHLH) protein family, in which the HLH domain is responsible for dimerization and the adjacent basic region mediates sequence-specific DNA binding. Myc has generally been associated with the promotion of cellular growth and proliferation, desensitization to growth-inhibitory stimuli, blockade of cell differentiation, cellular immortalization, and oncogenic transformation, as well as sensitization to apoptosis-inducing signals. Myc has also been linked to the regulation of various metabolic pathways, the induction of DNA damage, and genomic instability. In normal cells, c-Myc expression is generally induced by mitogens and suppressed by growth-inhibitory signals. Oncogenic activation of c-Myc occurs by direct gene alterations, such as translocation or amplification, or by mutations in upstream signaling pathways. These accidents commonly result in deregulated and/or elevated expression of c-Myc and its product. The activated c-Myc protein regulates multiple targeted genes through binding at the consensus DNA sequence CACGTG or E box.
- 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
- 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
- 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
- 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.
Transcription factor assay of c-Myc from nuclear extracts of Jurkat cells or U937 cells. A. Western-blot result of c-Myc from cytoplasm and nuclear fractions. B. Transcription factor assay of c-Myc from nuclear fractions with the RayBio® c-Myc TF Activity Assay.
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Transcription factor assay of c-Myc from nuclear extracts of Jurkat cells or U937 cells with the specific competitor or non-specific competitor. The result shows specific binding of c-Myc to the DNA binding site.
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.