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Protein-human TLK1B-Kinase domain

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Protein-human TLK1B-Kinase domain

TLK1B-Kinase domain

Protein name: Human TLK1-isoform 3-kinase domain (TLK1B-KD)
Tag name: 6X-Histidine
Expression system: E. Coli RGpLysS
Accession number: Q9UKI8-3

Recombinant TLK1B-KD amino acid sequence

MGSSHHHHHHSQDPKDHPTLNERYLLLHLLGRGGFSEVYKAFDLYEQRYAAVKIHQLNKSWRDEKKENYHKHACREYRIHKELDHPRIVKLYDYFSLDTDTFCTVLEYCEGNDLDFYLKQHKLMSEKEARSIVMQIVNALRYLNEIKPPIIHYDLKPGNILLVDGTACGEIKITDFGLSKIMDDDSYGVDGMDLTSQGAGTYWYLPPECFVVGKEPPKISNKVDVWSVGVIFFQCLYGRKPFGHNQSQQDILQENTILKATEVQFPVKPVVSSEAKAFIRRCLAYRKEDRFDVHQLANDPYLLPHMRRSNSSGNL*

Recombinant TLK1B-KD nucleotide sequence

ATGGGCAGCAGCCATCACCATCATCACCACAGCCAGGATCCAAAAGATCATCCGACCCTGAACGAACGCTATCTGCTGCTGCATCTGCTGGGCCGCGGCGGCTTTAGCGAAGTGTATAAAGCGTTTGATCTGTATGAACAGCGCTATGCGGCGGTGAAAATTCATCAGCTGAACAAAAGCTGGCGCGATGAAAAAAAAGAAAACTATCATAAACATGCGTGCCGCGAATATCGCATTCATAAAGAACTGGATCATCCGCGCATTGTGAAACTGTATGATTATTTTAGCCTGGATACCGATACCTTTTGCACCGTGCTGGAATATTGCGAAGGCAACGATCTGGATTTTTATCTGAAACAGCATAAACTGATGAGCGAAAAAGAAGCGCGCAGCATTGTGATGCAGATTGTGAACGCGCTGCGCTATCTGAACGAAATTAAACCGCCGATTATTCATTATGATCTGAAACCGGGCAACATTCTGCTGGTGGATGGCACCGCGTGCGGCGAAATTAAAATTACCGATTTTGGCCTGAGCAAAATTATGGATGATGATAGCTATGGCGTGGATGGCATGGATCTGACCAGCCAGGGCGCGGGCACCTATTGGTATCTGCCGCCGGAATGCTTTGTGGTGGGCAAAGAACCGCCGAAAATTAGCAACAAAGTGGATGTGTGGAGCGTGGGCGTGATTTTTTTTCAGTGCCTGTATGGCCGCAAACCGTTTGGCCATAACCAGAGCCAGCAGGATATTCTGCAGGAAAACACCATTCTGAAAGCGACCGAAGTGCAGTTTCCGGTGAAACCGGTGGTGAGCAGCGAAGCGAAAGCGTTTATTCGCCGCTGCCTGGCGTATCGCAAAGAAGATCGCTTTGATGTGCATCAGCTGGCGAACGATCCGTATCTGCTGCCGCATATGCGCCGCAGCAACAGCAGCGGCAACCTGTAA

Product Description:

Human TLK1B kinase domain protein expressed in Rosetta Gami PlysS strain of E.Coli, with N-terminal 6X-histidine tag, corresponding to MW=36.6 kDa Purity 95% by SDS-PAGE and Coomassie blue staining

Formulation: 1mg/ml of enzyme in 50 mM Tris-HCl pH 8.0, 150 mM NaCl, 0.2 mM TCEP, 2.5% Glycerol flash frozen in liquid N2 and stored at -80ºC.
Storage and Stability: Protein stored at -80 ºC for long-term use, stable at 4ºC for up to one week. The protein should be placed on ice during the usage once thawed.
Kinase assay protocol
1X Kinase reaction Buffer (KRB): 40 mM Tris-Cl (pH=7.5), 20 mM MgCl2, 2.5 % Glycerol, 0.1 mg/ml BSA
The desired concentration of TLK1B-KD (2X)- 2µM
Substrate dilution (2X): 2 µM in 1X KRB
ATP dilution (2X): 200 µM in 1X KRB

Assay Procedure:

1.The ADP-Glo kinase assay was performed in a solid, white, flat bottom 96-well plate.
2.The reaction volume of 20 µl with 2X 1 µM kinase TLK1B-KD, 2X 1 µM kinase substrate, and 2X 100 µM ATP in 1X Kinase Reaction Buffer (KRB).
3.The reaction was incubated for an hour at 30°C with gentle shaking.
4.20 µL of ADP-GLo reagent is added and incubated for 45 minutes.
5.40 µl of a Kinase Detection Reagent is added to each well and incubated for 60 minutes in the dark.
6.The readout was measured as luminescent values (Relative luminescent unit, RLU).

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