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164610
Sigma-Aldrichp-Aminophenylmercuric Acetate - CAS 6283-24-5 - Calbiochem
An organomercurial agent used for the activation of latent MMPs in vitro. Acts by facilitating the loss of the enzyme propeptide domain through an autolytic cleavage known as the cysteine switch.
More>>An organomercurial agent used for the activation of latent MMPs in vitro. Acts by facilitating the loss of the enzyme propeptide domain through an autolytic cleavage known as the cysteine switch. Less<<
MSDS (material safety data sheet) or SDS, CoA and CoQ, dossiers, brochures and other available documents.
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Description
Overview
An organomercurial agent used for the activation of latent MMPs in vitro. Acts by facilitating the loss of the enzyme propeptide domain through an autolytic cleavage known as the cysteine switch.
Note: 1 set = 5 x 700 mg.
Catalogue Number
164610
Brand Family
Calbiochem®
Synonyms
APMA
References
References
Chen, J.-M., et al. 1991. J. Biol. Chem. 266, 5113. Marcy, A.I., et al. 1991. Biochemistry30, 6476. Stricklin, G.P., et al. 1983. Biochemistry22, 61.
Very toxic by inhalation, in contact with skin and if swallowed. Irritating to eyes, respiratory system and skin.
S Phrase
S: 36
Wear suitable protective clothing.
Storage and Shipping Information
Ship Code
Ambient Temperature Only
Toxicity
Highly Toxic
Hazardous Materials Attention:
Due to the nature of the Hazardous Materials in this shipment, additional shipping charges may be applied to your order. Certain sizes may be exempt from the additional hazardous materials shipping charges. Please contact your local sales office for more information regarding these charges.
Storage
+15°C to +30°C
Do not freeze
Ok to freeze
Special Instructions
Following reconstitution, refrigerate (4°C). Stock solutions are stable for up to 1 month at 4°C.
Global Trade Item Number
Catalogue Number
GTIN
164610-700MG
04055977207637
Documentation
p-Aminophenylmercuric Acetate - CAS 6283-24-5 - Calbiochem Certificates of Analysis
Title
Lot Number
164610
References
Reference overview
Chen, J.-M., et al. 1991. J. Biol. Chem. 266, 5113. Marcy, A.I., et al. 1991. Biochemistry30, 6476. Stricklin, G.P., et al. 1983. Biochemistry22, 61.
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.
Revision
12-February-2024 JSW
Synonyms
APMA
Description
An organomercurial agent used for the activation of latent matrix metalloproteinases (MMPs) in vitro. Acts by facilitating the loss of the enzyme propeptide domain through an autolytic cleavage known as the cysteine switch.
Form
Yellow solid
Recommended reaction conditions
The MMPs are members of a family of at least twenty Zn2+-dependent endopeptidases that function extracellularly at neutral pH. The MMPs are initially synthesized as latent or inactive zymogens that contain a regulatory propeptide. This propeptide confers the latency by binding to the Zn2+ at the enzyme's active site. MMP activation can be achieved by autoactivation and by the action of other MMPs or proteases, such as furin, urokinase, plasmin, and trypsin, as well as by organomercurial compounds.
The following is a protocol from Stricklin, et al. (1983. Biochemistry22, 61) describing the use of paminophenylmercuric acetate (APMA) to activate pro-MMPs. This protocol is good for other types of organomercurials such as p-(hydroxymercuric) benzoate (PHMB), phenylmercuric chloride (PMC), and mersalyl.
Organomercurial stock solutions
Prepare stock solutions in 0.1 M NaOH at a concentration range of 10-50 mM just prior to use. Although not absolutely necessary, the stock solution may be adjusted to pH 11 with 5 N HCl. (e.g. Marcy, A.I. et al. 1991. Biochemistry30, 6476).
ProMMPs
Prepare proenzyme solutions in 0.1 M Tris-HCl, pH 7.5 at 1-2 mg/ml.
Activation of proMMPs
To initiate activation, combine proMMP solution with organomercurial stock in a 10:1 volume* ratio (MMPs to organomercurial). Incubate the mixture at 37°C for 2-3 hrs**. *When more organomercurial is desired, make the stock solution more concentrated. Do not exceed the above ratio so that the volume of organomercurial in the mixture is more than about 1/10 of that of the proMMP solution; this could result in significant changes in pH.
**It is recommended that an analytical run be conducted first to determine the optimal incubation time. For example, a small-scale experiment with fixed concentration of proMMP and organomercurial would be incubated as described above. Remove aliquots of sample at various time points. Stop the reaction by the addition of concentrated SDS-sample buffer (for example, add 10 µl of 2X sample buffer to a 10 µl aliquot or add 2 µl of 5X sample buffer to an 8 µl aliquot) and then boil. The progress of the activation can then be monitored qualitatively by analyzing these time aliquots on a 12% SDS-PAGE gel.
Removing organomercurial from the mixture after activation
The activated MMPs can be used without the removal of the organomercurial. Researchers are referred to the following reference for the removal of organomercurial by gel filtration if desired: Marcy, A.I., et al. 1991. Biochemistry30, 6476
CAS number
6283-24-5
RTECS
OV5550000
Chemical formula
C₈H₉HgNO₂
Structure formula
Purity
≥90% by HPLC
Solubility
DMSO (10 mg/ml) or 0.1 N NaOH (10 mg/ml)
Storage
+15°C to +30°C
Do Not Freeze
Ok to freeze
Special Instructions
Following reconstitution, refrigerate (4°C). Stock solutions are stable for up to 1 month at 4°C.
Toxicity
Highly Toxic
References
Chen, J.-M., et al. 1991. J. Biol. Chem. 266, 5113. Marcy, A.I., et al. 1991. Biochemistry30, 6476. Stricklin, G.P., et al. 1983. Biochemistry22, 61.
