Recombinant SUMO-GFP Protease Substrate Protein, CF

R&D Systems | Catalog # 11698-SO

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Key Product Details

  • R&D Systems E. coli-derived Recombinant SUMO-GFP Protease Substrate Protein (11698-SO)
  • Quality control testing to verify active proteins with lot specific assays by in-house scientists
  • All R&D Systems proteins are covered with a 100% guarantee

Source

E. coli

Accession Number

AAA58246

Applications

Enzyme Activity
View all SUMO-GFP Protease Substrate Proteins and Enzymes »

Product Specifications

Source

E. coli-derived crystal jellyfish SUMO-GFP Protease Substrate protein
Ser2-Lys238, with an N-terminal Met, 6-His tag, and Sumo tag

Purity

>95%, by SDS-PAGE visualized with Silver Staining and quantitative densitometry by Coomassie® Blue Staining.

Endotoxin Level

<1.0 EU per 1 μg of the protein by the LAL method.

N-terminal Sequence Analysis

Met

Predicted Molecular Mass

39 kDa

SDS-PAGE

42-47 kDa, under reducing conditions.

Activity

Measured by its cleavage by SUMO Protease ULP1.
>60% of 1 μg can be cleaved by 1 ng of SUMO Protease ULP1, as measured under the described conditions.

Scientific Data Images for Recombinant SUMO-GFP Protease Substrate Protein, CF

ULP1 and SENP1 cleavage of Recombinant SUMO-GFP Protease Substrate Protein.

Recombinant SUMO-GFP Protease Substrate (Catalog # 11698-SO) can be efficiently cleaved by SUMO Protease ULP1 (11697-SO) and Recombinant Human His6-SENP1 Catalytic Domain Protein (E-700).

Recombinant SUMO-GFP Protease Substrate Protein SDS-PAGE.

2 μg/lane of Recombinant SUMO-GFP Protease Substrate Protein (Catalog # 11698-SO) was resolved with SDS-PAGE under reducing (R) and non-reducing (NR) conditions and visualized by Coomassie® Blue staining, showing bands at 42-47 kDa, under reducing conditions.

Formulation, Preparation, and Storage

11698-SO
Formulation Supplied as a 0.2 μm filtered solution in Tris, NaCl and Glycerol.
Shipping The product is shipped with polar packs. Upon receipt, store it immediately at the temperature recommended below.
Stability & Storage Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
  • 6 months from date of receipt, -20 to -70 °C as supplied.
  • 3 months, -20 to -70 °C under sterile conditions after opening.

Background: SUMO-GFP Protease Substrate

SUMO-GFP Protease Substrate is a recombinant protein that contains SUMO fused to GFP that can be used as a positive control in an SDS-PAGE gel-shift assay to detect cleavage activity by Recombinant SUMO Protease ULP1 (Catalog # http://www.rndsystems.com/product_results.aspx?k=11697-SO">11697-SO) and Recombinant Human His6-SENP1 Catalytic Domain Protein
(Catalog # http://www.rndsystems.com/product_results.aspx?k=E-700">E-700). SUMO is a member of the small ubiquitin-like modifier (SUMO) family. In the cell, the SUMO protein is modified and covalently conjugated to other proteins in the cell via an isopeptide bond formed between its carboxyl-terminal glycine residues and the lysine of the protein substrate in a similar parallel mechanism of ubiquitination that regulates cellular processes such as DNA replication and repair, nuclear transport, signal transduction, and regulation of transcription (1-3). Use of SUMO fusions as a technology has been used to enhance protein expression, solubility, and purification in prokaryotes for a diverse population of protein targets and unlike other common fusion partners SUMO can serve as a chaperone for correct folding (4).  SUMO is cleaved at its c-terminus to a mature form after a glycine-glycine pair by a family of sentrin/SUMO-specific proteases (SENPs) in humans and ubiquitin-like protein specific proteases (Ulps) in yeast to regulate the reversible process of sumoylation (3). In a fusion application, the cleavage of SUMO allows for successful recombinant protein production with a native N-terminal sequence providing a distinct advantage over other fusion cleavage models used in recombinant protein production in both academic research and industrial biotechnology (4‑10).

References

  1. Li, S.J and M. Hochstrasser (1999) Nature 398:246.
  2. Li, S.J and M. Hochstrasser (2000) Mol. Cell Biol. 20:2367.
  3. Mukhopadhyay, D. and M. Dasso. (2007) Trends Biochem. Sci. 32:286.
  4. Malakhov, M.P. et al. (2004) J. Struct. Funct. Genomics 5:75.  
  5. Marblestone, J.G. et al. (2006) Protein Sci. 15:182.
  6. Baker, R.T. (1996) Curr. Opin. Biotechnol. 7:541.
  7. Jonasson, P. et al. (2002) Biotechnol. Appl. Biochem. 35:91.
  8. Butt, T.R. et al. (2005) Protein Expr. Purif. 43:1.
  9. Peroutka, R.J. et al. (2011) Methods Mol. Biol. 705:15.
  10. Wang, Z. et al. (2010) Protein Expr. Purif. 73:203.

Alternate Names

green fluorescent protein, small ubiquitin-related modifier, SUMO substrate

UniProt

AAA58246

Additional SUMO-GFP Protease Substrate Products

Product Documents for Recombinant SUMO-GFP Protease Substrate Protein, CF

Certificate of Analysis

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Product Specific Notices for Recombinant SUMO-GFP Protease Substrate Protein, CF

For research use only

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Protocols

View specific protocols for Recombinant SUMO-GFP Protease Substrate Protein, CF (11698-SO):

Materials
  • Assay Buffer: 50 mM Tris, 5 mM DTT, pH 8.0 
  • Recombinant SUMO-GFP Protease Substrate (rSUMO-GFP) (Catalog # 11698-SO) 
  • Recombinant Yeast SUMO Protease ULP1 His-tag (SUMO Protease ULP1) (Catalog # 11697-SO) 
  • 15% SDS-PAGE gel
  • Reducing Sample Buffer
  • Gel Staining Reagent
  1. Dilute rSUMO-GFP to 200 µg/mL in Assay Buffer.
  2. Dilute SUMO Protease ULP1 to 0.2 µg/mL in Assay buffer.
  3. Combine 20 µL of 0.2 µg/mL SUMO Protease ULP1and 20 µL of 200 µg/mL rSUMO-GFP. Prepare a Negative Control by combining 20 µL of 200 µg/mL rSUMO-GFP and 20 µL Assay Buffer.
  4. Incubate reaction mixtures and control at room temperature for 2 hours.
  5. Stop the reactions by combining equal volumes of reactions (including control) and Reducing SDS-PAGE Sample Buffer. Heat all samples at 95 °C for 3 minutes.
  6. Load 20 µL of each stopped reaction per lane on a 15% SDS-PAGE gel and perform electrophoresis.
  7. Stain gel and analyze the % cleavage of rSUMO-GFP using densitometry.

Per Well:

  • rSUMO-GFP: 1.0 µg
  • SUMO Protease ULP1: 1.0 ng 

FAQs

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