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Recombinant TBP protein
| Catalog No: 81114 | Format: 20 µg | ¥4,040 | Add to Cart |
| Catalog No: 81814 | Format: 1 mg | ¥33,200 | Add to Cart |
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Expressed In: Baculovirus Protein Species: Human
Contents
A representative Technical Data Sheet (TDS) is provided here. Please refer to the lot-specific TDS you will receive with your order for the lot-specific buffer contents and protein concentration.
Background
TBP (also known as TATA box binding protein, GTF2D, transcription initiation factor TFIID TBP subunit, TATA-binding factor, TF2D, TFIID or SCA17). RNA polymerase II is an enzyme found in eukaryotic cells composed of 12 subunits that catalyzes transcription of DNA. Transcription consists of three phases: initiation, elongation and termination. Transcription initiation involves TFIID, which is composed of TATA-binding protein (TBP) and TBP-associated factors (TAFs), as part of the RNA polymerase II pre-initiation complex (PIC). TBP binds to the TATA box sequence, which is located upstream of the transcription start site in some eukaryotic gene promoters, and helps position RNA polymerase II. TBP contains a string of glutamines at the N-terminus which affect DNA binding and the rate of transcription complex formation. TBP is also associated with RNA polymerase I and RNA polymerase III.
Application Notes
Application Notes: This protein is suitable for use in protein-protein interaction, in vitro transcription assay, binding assay.
Binding Assay conditions: 1 µM oligo dsDNA (DNA sequence: 5’-TCGTATAAAAGGC-3’) was incubated with different concentrations of TBP protein in 10 µl system containing 50 mM HEPES-NaOH pH 7.5, 0.1% BSA for 1 hour, then 10 µl FLAG antibody and SA-XL665 mixture (each 1:100 dilution in binding buffer) was added to each reaction system and incubated for 30 min. All the operations and reactions were performed at room temperature. HTRF assay was used for detection.
Protein Details
Recombinant human TBP protein was expressed in a baculovirus expression system as the full length protein (accession number NP_003185.1) with an N-terminal FLAG tag. The molecular weight of the protein is 39 kDa.
References
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Recombinant TBP protein
10% SDS-PAGE Coomassie staining
MW: 39 kDa
Purity: >80%
HTRF assay for TBP activity
1 µM oligo dsDNA (DNA sequence: 5’-TCGTATAAAAGGC-3’) was incubated with different concentrations of TBP protein in 10 µl system containing 50 mM HEPES-NaOH pH 7.5, 0.1% BSA for 1 hour, then 10 µl anti-FLAG antibody and SA-XL665 mixture (each 1:100 dilution in binding buffer) was added to each reaction system and incubated for 30 min. All the operations and reactions were performed at room temperature. HTRF assay was used for detection.
HTRF assay for TBP activity
1 µM oligo dsDNA (DNA sequence: 5’-TCGTATAAAAGGC-3’) was incubated with different concentrations of TBP protein in 10 µl system containing 50 mM HEPES-NaOH pH 7.5, 0.1% BSA for 1 hour, then 10 µl anti-FLAG antibody and SA-XL665 mixture (each 1:100 dilution in binding buffer) was added to each reaction system and incubated for 30 min. All the operations and reactions were performed at room temperature. HTRF assay was used for detection.
Storage
Recombinant proteins in solution are temperature sensitive and must be stored at -80°C to prevent degradation. Avoid repeated freeze/thaw cycles and keep on ice when not in storage.
Guarantee
This product is for research use only and is not for use in diagnostic procedures. This product is guaranteed for 6 months from date of arrival.
