GLPBio

Z-Arg-Arg-AMC

Catalog No.: GA23711Purity:>98.00%
COASDSData Sheet
Z-Arg-Arg-AMC is a specific substrate for histone B with an Ex/Em of 360/460nm.

Products are for research use only. Not for human use. We do not sell to patients.

Z-Arg-Arg-AMC
Cas No.: 88937-61-5
50mgIn Stock
$386.00
1
250mgIn Stock
$1,244.00
1
Tel: (909) 407-4943 / (909) 689-7124Email: sales@glpbio.com

Citations

2 publicationsPublications from researchers using this product

Description

Z-Arg-Arg-AMC is a specific substrate for histone B with an Ex/Em of 360/460nm. Z-Arg-Arg-AMC has pH-dependent cleavage properties.At neutral pH, Z-Arg-Arg-AMC is readily cleaved by histone B. However, Z-Arg-Arg-AMC is very poorly active at acidic pH with only one-third of the activity at neutral pH [1-4].

References:
[1] Ma C, Liang K, Tang L, He S, Liu X, He M, Li Y. Identification and characteristics of a cathepsin L-like cysteine protease from Clonorchis sinensis. Parasitol Res. 2019 Mar;118(3):829-835.
[2] Yoon MC, Phan V, Podvin S, Mosier C, O'Donoghue AJ, Hook V. Distinct Cleavage Properties of Cathepsin B Compared to Cysteine Cathepsins Enable the Design and Validation of a Specific Substrate for Cathepsin B over a Broad pH Range. Biochemistry. 2023 Aug 1;62(15):2289-2300.
[3] Yoon M C, Hook V, O’Donoghue A J. Cathepsin B dipeptidyl carboxypeptidase and endopeptidase activities demonstrated across a broad pH range[J]. Biochemistry, 2022, 61(17): 1904-1914.
[4] Klemenčič M, Novinec M, Dolinar M. Orthocaspases are proteolytically active prokaryotic caspase homologues: the case of M icrocystis aeruginosa[J]. Molecular Microbiology, 2015, 98(1): 142-150.

Protocol

This scheme is from the literature, please modify it according to your specific needs.

1. Solution configuration:

(1) Use anhydrous DMSO to dissolve Z-Arg-Arg-AMC to 100mM, and store in small portions at -20℃ to avoid repeated freeze-thaw.

(2) Use PAB (Hank's equilibrium salt solution does not contain sodium bicarbonate, contains 0.6mM CaCl2, 0.6mM MgCl2, 2 mm L-cysteine, 25 mm Pipes) to dilute Z-Arg-Arg-AMC with a concentration of 5-100μM.

Note: Please adjust the concentration of the working liquid according to the actual situation.

2. Cellular assay for cathepsin B activity

(1) Spread cells in 96-well plates and grow to 80% confluence (10-20,000 cells per well).

(2) Media was aspirated and cells were washed with 0.1mL Dulbecco's NaCl/Pi.

(3) Incubate cells with 0.2mL PAB for 30 minutes at 37°C.

(4) The buffer was replaced with fresh PAB containing 100µM Z-Arg-Arg-AMC, and cellular tissue protease B activity was determined in the presence or absence of 0.1% Triton X-100 (which increases the hydrolytic activity of Z-Arg-Arg-AMC).

(5) Detection was performed using a Labsystems Fluoroskan II microplate reader in kinetic mode at 37°C using a 360nm excitation filter and a 460nm emission filter.

Reference:

[1] Hulkower KI, Butler CC, Linebaugh BE, Klaus JL, Keppler D, Giranda VL, Sloane BF. Fluorescent microplate assay for cancer cell-associated cathepsin B. Eur J Biochem. 2000 Jul;267(13):4165-70.

Product Documents

Purity:>98.00%Appearance:A solid
COASDSData Sheet

Chemical Properties

CAS No.
88937-61-5
Molecular Formula
C30H39N9O6
Molecular Weight
621.7
Solubility
Soluble in DMSO
Storage
Store at -20°C
General tips
Select a suitable solvent according to the product solubility data. Once prepared, aliquot the stock solution to avoid repeated freeze-thaw cycles.
Use stock solutions within 6 months when stored at -80°C or within 1 month when stored at -20°C.
To improve solubility, warm the tube to 37°C and sonicate it for an appropriate period.
Shipping Condition
Evaluation samples are shipped with blue ice. Other available sizes are shipped at room temperature or with blue ice upon request.

CalculatorsMolarity / Dilution / Molecular Weight / Unit Conversion / In Vivo Formulation / Solubility

g/mol

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