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Recombinant SETD1B-SET complex
| Aliases: Histone-lysine N-methyltransferase SETD1B |
| Catalog No: 81342 | Format: 20 µg | ¥3,840 | Add to Cart |
| Catalog No: 81642 | 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
SETD1B (SET Domain Containing 1B, Histone Lysine Methyltransferase) is a component of a histone methyltransferase (HMT) complex (contain SETD1B-SET ASH2L, RBBP5, WDR5) that produces mono-, di-, and trimethylated histone H3 at Lys4, but not if the neighboring ‘Lys-9’ residue is already methylated. Trimethylation of histone H3 at lysine 4 (H3K4me3) is a chromatin modification known to generally mark the transcription start sites of active genes. The protein contains SET domains, a RNA recognition motif domain and is a member of the class V-like SAM-binding methyltransferase superfamily.
Application Notes
This complex is suitable for use in protein-protein interaction, in vitro transcription assay, binding assays.
Protein Details
Recombinant SETD1B-SET Complex that includes amino acids 1669-1966 of human SETD1B protein (accession number NP_001340274.1) with a N-terminal FLAG tag and full length human WDR5 protein (accession number NP_060058.1) without tag and full length human RBBP5 protein (accession number NP_005048.2) without tag and full length human ASH2L protein (accession number NP_001098684.1) without tag was expressed in Sf9 cells. The molecular weights of SETD1A-SET, WDR5, RBBP5,ASH2L are 35.7 kDa, 36.6 kDa, 59.2 kDa and 68.7 kDa, respectively.
Recombinant SETD1B-SET Complex
10% SDS-PAGE with Coomassie blue staining
MW of SETD1B-SET: 35.7 kDa
MW of WDR5: 36.6 kDa
MW of RBBP5: 59.2 kDa
MW of ASH2L: 68.7 kDa
Purity: >90%
HTRF assay for SETD1B-SET Complex activity
1 µM H3 (1-21) was incubated with different concentrations of SETD1B-SET Complex in a 10 µl reaction system containing 50 mM Tris-HCl pH 8.6, 0.02% Triton X-100, 2 mM MgCl2, 1 mM TCEP and 100 µM SAM for 2 hour, then 10 µl H3K4me2 antibody and SA-XL665 mixture (1:100 dilution in the same 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 SETD1B-SET Complex activity
1 µM H3 (1-21) was incubated with different concentrations of SETD1B-SET Complex in a 10 µl reaction system containing 50 mM Tris-HCl pH 8.6, 0.02% Triton X-100, 2 mM MgCl2, 1 mM TCEP and 100 µM SAM for 2 hour, then 10 µl H3K4me2 antibody and SA-XL665 mixture (1:100 dilution in the same 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.
