20S、26Sプロテアソーム、COP9シグナロソーム、プロテアソーム基質

 

 

 

 

 

 

 

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20S Proteasome (Human)

Quantity: 50 μg, 0.5mg/ml
Buffer: 20 mM Tris-HCl pH 7.2, 10% Glycerol, 150 mM KCl, 1 mM β–mercaptoethanol.
Source: Human red blood cells
Storage: -80o C. Avoid repeated freeze/thaw cycles

 

 

 

 

 

20S proteasome was purified from human blood cells by DEAE chromatography follow by the ammonium sulfate precipitation (60%), Mono Q and Resource Q chromatography. Purified complex was resolved on 10% SDS polyacrylamide gel and stained with Coomassie blue.

 

 

 

 

 

 

 

The chymotrypsin-like activity of the 20S proteasome (45μg/ml) was measured using 100μM Suc-LLVY-AMC in 20 mM HEPEs, pH 7.5, 0.5 mM EDTA, 0.05% Triton X, plus or minus 0.035% SDS

The chymotrypsin-like activity of the 20S proteasome (18 μg/ml) was measured using 100μM Suc-LLVY-AMC in 20 mM HEPEs, pH 7.5, 0.5 mM EDTA, 0.05% Triton X, plus or minus 0.5 mM MG132 (Cat. # SI9710)

Product Name	Cat #	Vol.
20S Proteasome (Human)	PS020	50 μg (0.5mg/ml)

 

26S Proteasome (Human)

Quantity: 25μg, 50μg, 0.5mg/ml
Buffer: 20 mM Tris-HCl pH 7.2, 10% Glycerol, 150 mM KCl, 1 mM β–mercaptoethanol.
Source: Human red blood cells
Storage: -80oC. Thaw the vial on ice. Avoid repeated freeze/thaw cycles

 

 

 

 

 

 

 

26S was purified from human blood cells by DEAE chromatography follow by the ammonium sulfate precipitation (60%), Mono Q and Resource Q chromatography. Purified complex was resolved on 10% SDS polyacrylamide gel and stained with Coomassie blue.

 

 

 

 

 

 

The chymotrypsin-like activity of the 26S proteasome (45μg/ml) was measured using 100μM Suc-LLVY-AMC in 20 mM HEPEs, pH 7.5, 0.5 mM EDTA, 0.05% Triton X, plus or minus 0.035% SDS

The chymotrypsin-like activity of the 26S proteasome (18 μg/ml) was measured using 100μM Suc-LLVY-AMC in 20 mM HEPEs, pH 7.5, 0.5 mM EDTA, 0.05% Triton X, plus or minus 0.5 mM MG132 (Cat. # SI9710)

Product Name	Cat #	Vol.
26S Proteasome (Human)	PS026	25μg (0.5mg/ml)
		50μg (0.5mg/ml)

 

 

COP9 Signalosome Complex (CSN, Human)

Quantity: 10 μg
Buffer: 20 mM Tris-HCl pH 7.2, 10% Glycerol, 150 mM KCl, 1 mM β–mercaptoethanol.
Source: Human red blood cells
Storage: -80oC. Avoid repeated freeze/thaw cycles

 

 

 

CSN complex was purified to homogeneity from human red blood cells using DEAE, Mono Q and Resource Q chromatography. Final fraction was resolved on 10% SDS polyacrylamide gel and stained with Coomassie blue (A). This preparation retains deneddylase activity (B)

 

Product Name	Cat #	Vol.
COP9 Signalosome Complex (CSN, Human)	CP009	10 μg

 

 

Rpn1 Protein (26S Proteasome Regulatory Subunit 1)

Quantity: 50 μg
Molecular weight: 109.5kDa
Tag: His6
Buffer: 20 mM sodium phosphate pH 7.4, 500 mM NaCl, 200 mM imidazole, 1mM β– mercaptoethanol.
Source: Yeast recombinant, expressed in E. coli. Accession number P38764.
Storage: -80oC. Avoid repeated freeze/thaw cycles

 

 

 

 

 

 

Recombinant Rpn1 protein (Hisx6 tag) was expressed in E.coli and purified on the Ni-NTA resin using a standard protocol. 5ug of purified protein was resolved on 10% PAGE and stained with Coomassie blue.

 

 

 

 

Product Name	Cat #	Vol.
Rpn1 Protein (26S Proteasome Regulatory Subunit 1)	PS110	50 μg

 

 

Rpn2 Protein (26S Proteasome Regulatory Subunit 2)

Quantity: 50 μg
Molecular weight: 104.2kDa
Tag: His6
Buffer: 20 mM sodium phosphate pH 7.4, 500 mM NaCl, 200 mM imidazole, 1mM β– mercaptoethanol.
Source: Yeast recombinant, expressed in E. coli. Accession number P32565.
Storage: -80oC. Avoid repeated freeze/thaw cycles

 

 

 

 

Recombinant Rpn2 protein (Hisx6 tag) was expressed in E.coli and purified on the Ni-NTA resin using a standard protocol. 5ug of purified protein was resolved on 10% PAGE and stained with Coomassie blue.

 

 

 

 

 

Product Name	Cat #	Vol.
Rpn2 Protein (26S Proteasome Regulatory Subunit 2)	PS120	50 μg

 

 

Suc-Leu-Leu-Val-Tyr-AMC (Suc-LLVY-AMC), Proteasome Substrate

Quantity: 5mg
Molecular weight: 763.9
Formula: C40H53N5O10
State: Lyophilized powder
Purity: >98% by TLC
Solubility: DMSO at least 5mM. Concentration range 10-100μM.
Storage: Dry reagent at 4oC; Store DMSO stock at -20oC. Avoid repeated freeze/thaw cycles

Formula:

 

 

 

 

 

 

 

 

 

Product Name	Cat #	Vol.
Suc-Leu-Leu-Val-Tyr-AMC (Suc-LLVY-AMC), Proteasome Substrate	PS500	5mg

 

 

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