530032 Nedd4/Rsp5 Signaling Inducer, NAB2 - Calbiochem

530032
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Overview

Replacement Information

Key Specifications Table

Empirical Formula
C₂₃H₂₀ClN₃O

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      Glass bottle 10 mg
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      Description
      OverviewA cell-permeable N-aryl benzimidazole (NAB) compound that protects against α-synuclein (α-syn) toxicity in yeast (0.625 to 10 µM), wt human α-syn-expressing C. elegans dopaminergic neurons (10 µM), human A53T α-syn-expressing rat DA neurons (0.1 µM), as well as in Parkinson's disease patients-derived neurons carrying AT53T α-syn or an α-syn triplication (5 to 40 µM) by restoring Nedd4 (mammalian)/Rsp5 (yeast) E3 ligase-dependent trafficking events, reducing α-syn-caused nitrosative and ER stress without affecting cellular α-syn level or Nedd4/Rsp5 ligase activity.
      Catalogue Number530032
      Brand Family Calbiochem®
      SynonymsN-(2-Chlorobenzyl)-1-(2,5-dimethylphenyl)-1H-benzo[d]imidazole-5-carboxamide
      References
      ReferencesTardiff, D.F., et al. 2013. Science 342, 979.
      Chung, C.Y., et al. 2013. Science 342, 983.
      Product Information
      FormOff-white solid
      Hill FormulaC₂₃H₂₀ClN₃O
      Chemical formulaC₂₃H₂₀ClN₃O
      ReversibleY
      Quality LevelMQ100
      Applications
      Biological Information
      Purity≥98% by HPLC
      Physicochemical Information
      Cell permeableY
      Dimensions
      Materials Information
      Toxicological Information
      Safety Information according to GHS
      Safety Information
      Product Usage Statements
      Storage and Shipping Information
      Ship Code Ambient Temperature Only
      Toxicity Standard Handling
      Storage +2°C to +8°C
      Protect from Light Protect from light
      Do not freeze Ok to freeze
      Special InstructionsFollowing reconstitution, aliquot and freeze (-20°C). Stock solutions are stable for up to 6 months at -20°C.
      Packaging Information
      Packaged under inert gas Packaged under inert gas
      Transport Information
      Supplemental Information
      Specifications

      Documentation

      Nedd4/Rsp5 Signaling Inducer, NAB2 - Calbiochem SDS

      Title

      Safety Data Sheet (SDS) 

      References

      Reference overview
      Tardiff, D.F., et al. 2013. Science 342, 979.
      Chung, C.Y., et al. 2013. Science 342, 983.
      Data Sheet

      Note that this data sheet is not lot-specific and is representative of the current specifications for this product. Please consult the vial label and the certificate of analysis for information on specific lots. Also note that shipping conditions may differ from storage conditions.

      Revision18-April-2014 JSW
      SynonymsN-(2-Chlorobenzyl)-1-(2,5-dimethylphenyl)-1H-benzo[d]imidazole-5-carboxamide
      DescriptionA cell-permeable N-aryl benzimidazole (NAB) compound that protects against α-synuclein (α-syn) toxicity in yeast (0.625 to 10 µM), wt human α-syn-expressing C. elegans dopaminergic (DA) neurons (10 µM), human A53T α-syn-expressing rat DA neurons (0.1 µM), as well as in human neurons differentiated from iPSCs derived from Parkinson′s disease (PD) patients carrying either AT53T α-syn or an α-syn triplication (5 to 40 µM) by restoring Nedd4 (mammalian)/Rsp5 (yeast) E3 ligase-dependent processes, including ER-to-Golgi, plasma membrane- and Golgi-to-vacuole/lysosome trafficking events, reducing α-syn-caused nitrosative and ER stress. NAB2 is also reported to inhibit the growth, but not viability, of non-α-syn-expressing yeast via the same mechanism of action (MOA) albeit at a lower potency (GI40 = 20.5 µM vs. EC40 = 4.5 µM against α-syn toxicity). Although the exact molecular mechanism is yet unknown, NAB2 does not affect cellular α-syn level or Nedd4/Rsp5 ligase activity.
      FormOff-white solid
      Intert gas (Yes/No) Packaged under inert gas
      Recommended reaction conditions
      Direct dilution of NAB2:
      1. Dissolve powdered NAB2 at 10 mM in 100% DMSO
      2. When adding to cells, do not pipette NAB2 directly to media.
      3. Add volume of NAB2 stock to empty tube to which media can be added. The volume should be chosen such that your final dilution will yield a final concentration within the recommended testing range of 1 to 40 µM.
      4. Add media to this tube while gently vortexing sto yield a final NAB2 concentration less than or equal to 40 µM.
      5. Apply to cells.
      6. Note if media appears cloudy or if particulates are present. If so, discard and dilute NAB2 to a lower concentration. 7. We also recommend simultaneously using the alternative protocol below.

      Alternative protocol: Using NAB2-solubilizing agent.
      1. Dissolve powdered NAB2 at 10 mM in 100% DMSO. 2. Make a stock solution of (2-Hydroxypropyl)-β-cyclodextrin at 33% in water (% w/v). This compound possesses a ring-like structure with a hydrophobic central cavity and hydrophilic exterior that increases the solubility of hydrophobic compounds through well described host-guest complex formation that increases bioavailability.
      3. Add NAB2 (10 mM DMSO stock) to empty tube. Add a 4-fold volume of 33% (% w/v) (2-Hydroxypropyl)-β-cyclodextrin. Vortex and let sit for 10 minutes. NAB2 concentration will now by 2 mM. This is your stock solution. It can be stored at 4°C for 1 month or lyophilized and frozen long-term storage.
      4. Add desired volume of NAB2:cyclodextrin complex to empty tube to which media can be added. The volume should be chosen such that your final dilution will yield a NAB2 concentration over the recommended testing range of 1 to 40 µM.
      4. Add media to this tube such that the final NAB2 concentration does not exceed 40 µM.
      5. Apply to cells.
      Chemical formulaC₂₃H₂₀ClN₃O
      Purity≥98% by HPLC
      SolubilityDMSO (100 mg/ml)
      NAB2 is moderately hydrophobic (clogP ~5.2)and can precipitate out of solution if added directly to cell culture or yeast media. Care should therefore be taken to optimize solubility and guard against false negative results caused by irrecoverable precipitation of the compound. Please refer to the Recommended Reaction Conditions above for details.
      Storage +2°C to +8°C
      Protect from light
      Do Not Freeze Ok to freeze
      Special InstructionsFollowing reconstitution, aliquot and freeze (-20°C). Stock solutions are stable for up to 6 months at -20°C.
      Toxicity Standard Handling
      ReferencesTardiff, D.F., et al. 2013. Science 342, 979.
      Chung, C.Y., et al. 2013. Science 342, 983.