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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      Catalog Number AvailabilityPackaging Qty/Pack Price Quantity
      5.30032.0001
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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
          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.