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  4. Cys(Npys)-(Arg)9

Cys(Npys)-(Arg)9 is a synthetic cationic cell-penetrating peptide with a reversible thiol-reactive nitropyridyl (Npys) group. Cys(Npys)-(Arg)9 efficiently mediates the internalization and delivery of various "cargo" such as proteins and antibodies by forming reversible disulfide bonds with surface-exposed cysteine residues. Cys(Npys)-(Arg)9 endows TALEN proteins with cell-penetrating activity, enabling gene knockout in mammalian cells and protein transduction in wheat microspores. Cys(Npys)-(Arg)9 can be conjugated with antibodies to form cationized IgG for enhancing endosomal escape of oligonucleotides, or form siRNA delivery complexes. When the molar ratio of Cys(Npys)-(Arg)9 to loaded molecules is higher than 1:1, it exerts certain cytotoxic effects on cells. Cys(Npys)-(Arg)9 can be used in studies related to oral squamous cell carcinoma and HIV infection.

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Cys(Npys)-(Arg)9

Cys(Npys)-(Arg)9 Chemical Structure

CAS No. : 1417221-06-7

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Cys(Npys)-(Arg)9 Related Antibodies

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Description

Cys(Npys)-(Arg)9 is a synthetic cationic cell-penetrating peptide with a reversible thiol-reactive nitropyridyl (Npys) group. Cys(Npys)-(Arg)9 efficiently mediates the internalization and delivery of various "cargo" such as proteins and antibodies by forming reversible disulfide bonds with surface-exposed cysteine residues. Cys(Npys)-(Arg)9 endows TALEN proteins with cell-penetrating activity, enabling gene knockout in mammalian cells and protein transduction in wheat microspores. Cys(Npys)-(Arg)9 can be conjugated with antibodies to form cationized IgG for enhancing endosomal escape of oligonucleotides, or form siRNA delivery complexes. When the molar ratio of Cys(Npys)-(Arg)9 to loaded molecules is higher than 1:1, it exerts certain cytotoxic effects on cells. Cys(Npys)-(Arg)9 can be used in studies related to oral squamous cell carcinoma and HIV infection[1][2][3][4][5].

In Vitro

Upon conjugation with CCR5-targeting TALEN protein, Cys(Npys)-(Arg)9 induces CCR5 gene knockout in HeLa cells at a frequency of approximately 16% when delivered at a peptide-to-protein ratio of 8:1 or 15:1 (1 μM; 2 hr)[1].
After conjugation of Cys(Npys)-(Arg)9 to TALEN proteins targeting BMPR1A, delivery at peptide-to-protein ratios of 1:1, 2:1 or 4:1 (1 μM; 2 h) induces knockout of the BMPR1A gene in HEK 293 cells, with a maximum frequency of up to 4%[1].
Cys(Npys)-(Arg)9-conjugated TALEN proteins do not reduce the viability of HeLa cells or HEK 293 cells[1].

MedChemExpress (MCE) has not independently confirmed the accuracy of these methods. They are for reference only.

Cys(Npys)-(Arg)9 Related Antibodies

Cell Viability Assay[1]

Cell Line: HeLa and HEK 293 cells
Concentration: Various concentrations of conjugated TALEN protein
Incubation Time: 2 hr
Result: Showed no apparent reduction in cell viability in either HeLa or HEK 293 cells after treatment.
Demonstrated increased toxicity only associated with decreased TALEN protein purity, not the conjugated peptide itself.
Molecular Weight

1679.99

Formula

C62H118N40O12S2
CAS No.
Sequence

Cys(Nyps)-Arg-Arg-Arg-Arg-Arg-Arg-Arg-Arg-Arg-NH2
Sequence Shortening

C(Npys)RRRRRRRRR-NH2
Shipping

Room temperature in continental US; may vary elsewhere.
Storage

Please store the product under the recommended conditions in the Certificate of Analysis.
Solvent & Solubility
In Vitro: 

H2O

Peptide Solubility and Storage Guidelines:

1.  Calculate the length of the peptide.

2.  Calculate the overall charge of the entire peptide according to the following table:

  Contents Assign value
Acidic amino acid Asp (D), Glu (E), and the C-terminal -COOH. -1
Basic amino acid Arg (R), Lys (K), His (H), and the N-terminal -NH2 +1
Neutral amino acid Gly (G), Ala (A), Leu (L), Ile (I), Val (V), Cys (C), Met (M), Thr (T), Ser (S), Phe (F), Tyr (Y), Trp (W), Pro (P), Asn (N), Gln (Q) 0

3.  Recommended solution:

Overall charge of peptide Details
Negative (<0) 1.  Try to dissolve the peptide in water first.
2.  If water fails, add NH4OH (<50 μL).
3.  If the peptide still does not dissolve, add DMSO (50-100 μL) to solubilize the peptide.
Positive (>0) 1.  Try to dissolve the peptide in water first.
2.  If water fails, try dissolving the peptide in a 10%-30% acetic acid solution.
3.  If the peptide still does not dissolve, try dissolving the peptide in a small amount of DMSO.
Zero (=0) 1.  Try to dissolve the peptide in organic solvent (acetonitrile, methanol, etc.) first.
2.  For very hydrophobic peptides, try dissolving the peptide in a small amount of DMSO, and then dilute the solution with water to the desired concentration.
Purity & Documentation
References
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Cys(Npys)-(Arg)9 Related Classifications

  • Molarity Calculator

  • Dilution Calculator

The molarity calculator equation

Mass (g) = Concentration (mol/L) × Volume (L) × Molecular Weight (g/mol)

Mass   Concentration   Volume   Molecular Weight *
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The dilution calculator equation

Concentration (start) × Volume (start) = Concentration (final) × Volume (final)

This equation is commonly abbreviated as: C1V1 = C2V2

Concentration (start) × Volume (start) = Concentration (final) × Volume (final)
× = ×
C1   V1   C2   V2
Help & FAQs
  • Do most proteins show cross-species activity?

    Species cross-reactivity must be investigated individually for each product. Many human cytokines will produce a nice response in mouse cell lines, and many mouse proteins will show activity on human cells. Other proteins may have a lower specific activity when used in the opposite species.

Keywords:

Cys(Npys)-(Arg)91417221-06-7HIVHuman immunodeficiency virusanti-CD63 antibodyBMPR1AscFvCD7-CysHIV infectionoral squamous cell carcinomaCys-mCherrymammalian cellswheat microsporeshuman CCR5TALEN proteinsInhibitorinhibitorinhibit

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