Alkaline Sphingomyelinase Activity Assay Kit (Colorimetric) (BN01145)

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BN01145
  • Alkaline Sphingomyelinase Activity Assay Kit (Colorimetric) (BN01145)
  • Alkaline Sphingomyelinase Activity Assay Kit (Colorimetric) (BN01145)
  • Alkaline Sphingomyelinase Activity Assay Kit (Colorimetric) (BN01145)
$755.44

Description

ELISA Kit Technical ManualMSDS

Alkaline Sphingomyelinase Activity Assay Kit (Colorimetric)

I. Sphingomyelinase (SMase, E.C. 3.1.4.12) cleaves sphingomyelin to produce phosphocholine and ceramide. Both of these molecules can act as second messengers and regulate downstream events including apoptosis, cell proliferation, and sterol homeostasis. The sphingomyelinase family of enzymes includes acid sphingomyelinase, found in the lysosome and also secreted, neutral sphingomyelinase, which is enriched in the brain, and alkaline sphingomyelinase (AlkSMase), which acts in the intestinal tract. Unlike the neutral variety (nSMase), AlkSMase shows optimal activity at higher pH and does not require Mg2+ for enzymatic activity. AlkSMase, also referred to as NPP7, because of its homology to the Nucleotide Pyrophosphatase/Phosphodiesterase family of enzymes, has been implicated in prevention of tumor formation. Its activity is decreased in some intestinal cancers, as wells as instances of chronic colitis. Alkaline Sphingomyelinase may also play a role in cholesterol absorption. Assay Genie's Alkaline Sphingomyelinse Colorimetric Assay Kit provides a simple, high throughput adaptable means to identify and quantify alkaline sphingomyelinase activity in a variety of samples without influence from neutral or acid sphingomyelinase. Upon incubation with substrate, the hydrolysis produces an intermediate that reacts with the probe, generating a colorimetric signal. The assay can detect as low as 0.5 µU of alkaline sphingomyelinase activity

Figure: (a) Choline standard curve; (b) Alkaline Sphingomyelinase activity of NPP7. Hydrolysis was allowed to proceed for 1 hr. Neutral Sphingomyelinase (nSMase, purple bar) showed minimal activity after the same incubation time. (c) Activity determination of intestinal tissue lysate. For this experiment, 100 mg rat intestine was used, following Alkaline Sphingomyelinase Activity Assay Kit protocol with 2X protease inhibitor. Lysate was assayed and specific activity was determined to be 0.016 nmol/min/µg lysate.

Key Information Description

Product SKU

BN01145

Size

100 assays

Detection Method

Absorbance (OD 570 nm)

Applications

Sensitive assays for measuring Alkaline Sphingomyelinase activity in various biological samples and the kit is also suitable for high throughput analyses.

Features and Benefits

  • Simple, rapid & convenient
  • The assay can detect sphingomyleinase activity as low as 0.5 in a variety of samples.

Kit Components

  • AlkSMase Assay BufferAlkSMase Developer Buffer
  • AlkSMase Probe (DMSO)
  • AlkSMase Substrate (Lyophilized)
  • AlkSMase Enzyme Mix I (Lyophilized)
  • AlkSMase Enzyme Mix II (Lyophilized)
  • Choline Standard (Lyophilized)
  • Alk SMase Positive Control (Lyophilized)

Storage Conditions

-20°C

Shipping Conditions

Gel Pack

USAGE

For Research Use Only! Not For Use in Humans.

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