SARM1 Fluorogenic Assay Kit (Hydrolase Activity)
The SARM1 Fluorogenic Assay Kit (Hydrolase Activity) is designed to measure NAD+ cleavage activity in screening and profiling applications. The SARM1 assay kit comes in a convenient 96 or 384-well format, with enough recombinant human SARM1 enzyme (amino acids 28-724), its substrate N6-etheno-NAD (e-NAD), and SARM1 assay buffer for 100 or 400 enzyme reactions. In addition, the kit includes a SARM1 inhibitor (DSRM-3716) for use as a control inhibitor.
Figure 1: Assay principle.
Hydrolase activity of SARM1 is measured by following the hydrolysis of Etheno-NAD to form Etheno-ADPR + nicotinamide. Etheno-NAD is not fluorescent due to internal quenching. Upon hydrolysis by SARM1, nicotinamide is released, leading to an increase in fluorescence signal directly proportional to the enzymatic activity.
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- Adjustable micro-pipettor and sterile tips
- Fluorescent microplate reader able to excite at 300 nm and detection of emitted light at 410 nm.
- Rotating or rocker platform
96 reactions
| Catalog # | Name | Amount | Storage |
| 100069 | SARM1, FLAG-Tag Recombinant* | 30 µg | -80°C |
| 4x SARM1 Hydrolase Buffer | 3 ml | -20°C | |
| SARM1 Substrate (e-NAD, 12 mM) | 50 µl | -20°C | |
| DSRM-3716 (100 mM in DMSO) | 25 µl | -20°C | |
| 79685 | Black 96-well plate | 1 | Room Temp |
*The initial concentration of enzyme is lot-specific and will be indicated on the tube containing the protein.
384 reactions
| Catalog # | Name | Amount | Storage |
| 100069 | SARM1, FLAG-Tag Recombinant* | 2 x 30 µg | -80°C |
| 4x SARM1 Hydrolase Buffer | 2 x 3 ml | -20°C | |
| SARM1 Substrate (e-NAD, 12 mM) | 2 x 50 µl | -20°C | |
| DSRM-3716 (100 mM in DMSO) | 2 x 25 µl | -20°C | |
| 79961 | Black 384-well plate | 1 | Room Temp |
*The initial concentration of enzyme is lot-specific and will be indicated on the tube containing the protein.
SARM1 (Sterile alpha and TIR motif containing 1) is a member of the Toll/Interleukin receptor-1 (TIR1) family of enzymes. It functions as an ADP-ribosyl cyclase and nicotinamide adenine dinucleotide (NAD) glycohydrolase. SARM1-TIR domains have intrinsic NADase activity, cleaving NAD+ into ADP Ribose (ADPR), cyclic ADPR, and Nicotinamide. Often associated with mitochondria, the protein functions as a sensor of metabolic stress. It is highly expressed in neurons, where it causes the depletion of axonal NAD+ and pathological axon loss.
SARM1 functions downstream of NMNAT2 (nicotinamide nucleotide adenylyltransferase 2) to promote the active process of injury-induced neuronal degeneration known as Wallerian degeneration. Constitutive NADase activity resulting from mutation in the human SARM1 gene has been observed in neurodegenerative disease amyotrophic lateral sclerosis (ALS, or Lou Gehrig's disease). Alternatively, loss of SARM1 activity protects neurons in models of brain injury or drug-induced neuron damage. Therefore, inhibition of SARM1 NAD+ cleavage activity may potentially reduce axonal degeneration.
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