Protein-human TLK1B-full length
TLK1B-full length
Protein name: Human TLK1-isoform 3 (TLK1B)
Tag name: 6X-Histidine
Expression system: E. Coli RGpLysS
Accession number: Q9UKI8-3
Recombinant TLK1B amino acid sequence
MGSSHHHHHHSQDPMLKLAALESNKIQDLEKKEGRIDDLLRANCDLRRQIDEQQKLLEKYKERLNKCISMSKKLLIEKSTQEKLSSREKSMQDRLRLGHFTTVRHGASFTEQWTDGFAFQNLVKQQEWVNQQREDIERQRKLLAKRKPPTANNSQAPSTNSEPKQRKNKAVNGAENDPFVRPNLPQLLTLAEYHEQEEIFKLRLGHLKKEEAEIQAELERLERVRNLHIRELKRINNEDNSQFKDHPTLNERYLLLHLLGRGGFSEVYKAFDLYEQRYAAVKIHQLNKSWRDEKKENYHKHACREYRIHKELDHPRIVKLYDYFSLDTDTFCTVLEYCEGNDLDFYLKQHKLMSEKEARSIVMQIVNALRYLNEIKPPIIHYDLKPGNILLVDGTACGEIKITDFGLSKIMDDDSYGVDGMDLTSQGAGTYWYLPPECFVVGKEPPKISNKVDVWSVGVIFFQCLYGRKPFGHNQSQQDILQENTILKATEVQFPVKPVVSSEAKAFIRRCLAYRKEDRFDVHQLANDPYLLPHMRRSNSSGNLHMAGLTASPTPPSSSIITY*
Recombinant TLK1B amino acid sequence
ATGGGCAGCAGCCATCACCATCATCACCACAGCCAGGATCCAATGCTGAAACTGGCGGCGCTGGAGAGCAACAAGATCCAGGATCTGGAGAAGAAAGAAGGTCGTATTGACGATCTGCTGCGTGCGAACTGCGATCTGCGTCGTCAAATCGACGAACAGCAAAAACTGCTGGAGAAGTACAAAGAACGTCTGAACAAGTGCATCAGCATGAGCAAGAAACTGCTGATTGAGAAAAGCACCCAGGAAAAGCTGAGCAGCCGTGAGAAAAGCATGCAAGACCGTCTGCGTCTGGGTCACTTCACCACCGTGCGTCACGGCGCGAGCTTTACCGAGCAGTGGACCGATGGTTTCGCGTTTCAAAACCTGGTGAAACAGCAAGAATGGGTTAACCAGCAACGTGAGGACATTGAACGTCAGCGTAAGCTGCTGGCGAAGCGTAAACCGCCGACCGCGAACAACAGCCAGGCGCCGAGCACCAACAGCGAGCCGAAACAGCGTAAGAACAAAGCGGTGAACGGTGCGGAAAACGACCCGTTTGTTCGTCCGAACCTGCCGCAGCTGCTGACCCTGGCGGAGTATCACGAACAAGAGGAAATCTTCAAGCTGCGTCTGGGCCACCTGAAGAAAGAGGAAGCGGAGATTCAGGCGGAGCTGGAACGTCTGGAACGTGTTCGTAACCTGCACATCCGTGAGCTGAAACGTATTAACAACGAAGATAACAGCCAATTCAAGGACCACCCGACCCTGAACGAACGTTACCTGCTGCTGCACCTGCTGGGTCGTGGTGGCTTCAGCGAGGTGTATAAAGCGTTTGATCTGTACGAACAGCGTTATGCGGCGGTTAAAATCCACCAACTGAACAAGAGCTGGCGTGACGAGAAGAAAGAAAACTACCACAAGCACGCGTGCCGTGAGTATCGTATCCACAAAGAACTGGATCACCCGCGTATTGTGAAGCTGTACGACTATTTCAGCCTGGATACCGACACCTTTTGCACCGTTCTGGAGTACTGCGAAGGCAACGATCTGGACTTTTATCTGAAGCAGCACAAACTGATGAGCGAGAAAGAAGCGCGTAGCATCGTGATGCAAATTGTTAACGCGCTGCGTTACCTGAACGAGATCAAACCGCCGATCATTCACTATGATCTGAAGCCGGGTAACATTCTGCTGGTTGACGGCACCGCGTGCGGCGAAATCAAAATTACCGATTTCGGCCTGAGCAAGATCATGGACGATGACAGCTACGGTGTTGATGGTATGGACCTGACCAGCCAGGGTGCGGGCACCTACTGGTATCTGCCGCCGGAGTGCTTTGTGGTTGGCAAGGAACCGCCGAAAATCAGCAACAAGGTGGATGTTTGGAGCGTGGGCGTTATTTTCTTTCAGTGCCTGTATGGTCGTAAACCGTTCGGCCACAACCAGAGCCAGCAAGACATCCTGCAAGAGAACACCATTCTGAAGGCGACCGAAGTGCAATTCCCGGTTAAACCGGTGGTTAGCAGCGAGGCGAAGGCGTTTATCCGTCGTTGCCTGGCGTACCGTAAGGAAGATCGTTTCGACGTGCACCAGCTGGCGAACGACCCGTATCTGCTGCCGCACATGCGTCGTAGCAACAGCAGCGGTAACCTGCACATGGCGGGTCTGACCGCGAGCCCGACCCCGCCGAGCAGCAGCATCATTACCTACTAA
Product Description:
Human TLK1B full length expressed in Rosetta Gami PlysS strain of E.Coli, with N-terminal 6X-histidine tag, corresponding to MW=64.9 kDa Purity 85% by SDS-PAGE and Coomassie blue staining
Purity was assessed by SDS-PAGE and Coomassie staining of 1mg/ml TLK1B protein
Formulation: 1mg/ml of enzyme in 50 mM Tris-HCl pH 6.9, 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 (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, 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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