BLM Helicase Activity Assay Kit
The BLM Helicase Activity Assay Kit is a fluorogenic assay designed for screening and profiling of BLM (Bloom syndrome protein) antagonists/inhibitors by monitoring their inhibitory effects on BLM-catalyzed DNA unwinding. The BLM Helicase Activity Assay Kit comes in a convenient 96-well format, with enough purified recombinant BLM (amino acids 630-1300), ATP, DNA substrate, assay buffer, and additives for 100 reactions.
Figure 1: Illustration of the assay principle.
BLM is incubated with the fluorescent DNA substrate in the presence of ATP. The DNA substrate is conjugated on one strand with the TAMRA (tetramethylrhodamine) fluorophore, and on the other strand with BHQ (Black Hole Quencher) which effectively quenches TAMRA fluorescence due to their proximity within the DNA double strand. BLM unwinding of the DNA probe separates the two strands, releasing TAMRA fluorescence. BLM activity, therefore, results in a proportional increase in fluorescence.
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Fluorescent microplate reader capable of reading λex/em=525 nm/592 nm.
| Catalog # | Name | Amount | Storage |
| 102130 | BLM, GST-Tag* | 10 µg | -80°C |
| 4x BLM Buffer | 4 ml | -20°C | |
| DNA Substrate | 20 µl | -80°C | |
| 200 mM ATP | 50 µl | -20°C | |
| 0.5 M DTT | 200 ml | -20°C | |
| 79685 | Black, low binding plate | 1 | Room Temp |
| Plate cover | 1 | Room Temp |
*The concentration of the protein is lot-specific and will be indicated on the tube.
The BLM helicase, also known as Bloom syndrome protein, is a key enzyme involved in DNA replication and repair (DDR). The BLM helicase is a member of the RecQ family of helicases, which are evolutionarily conserved and found in many organisms, including bacteria, yeast, and humans. It catalyzes the unwinding of duplex DNA with 3’ to 5’ directionality, driven by the energy generated from ATP hydrolysis. BLM plays a crucial role in maintaining genomic stability by unwinding DNA structures during processes such as DNA replication, recombination, and repair. Mutations in the BLM gene can lead to Bloom syndrome, a rare genetic disorder characterized by growth deficiency, sun-sensitive skin lesions, and an increased risk of cancer. High expression of BLM is found in glioblastoma, and it was found that inhibition of its activity leads to increased susceptibility to treatment with drugs targeting other proteins involved in DDR, such as PARP1 (poly-ADP ribosylation protein 1). The use of BLM inhibitors as part of a combinatory therapeutic approach may open new avenues of treatment in cancer therapy.
Chen, X., et al., 2021. eLife 10 :e65339.
Wojnicki K., et al., 2023 Cell Death Discovery 9 :157.
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