MilliporeSigma
All Photos(2)

Documents

SAE0147

Sigma-Aldrich

Thrombin protease, biotin-tagged

human recombinant,expressed in HEK 293 cells, ≥5000 units/mL

Sign Into View Organizational & Contract Pricing

Synonym(s):
Factor IIa, fibrinogenase, thrombase

recombinant

expressed in HEK 293 cells

concentration

≥5000 units/mL

shipped in

dry ice

storage temp.

−20°C

Compare Similar Items

View Full Comparison

Show Differences

1 of 4

This Item
T932610602400001SAE0110
vibrant-m

SAE0147

Thrombin protease, biotin-tagged

vibrant-m

T9326

Thrombin human

vibrant-m

10602400001

Thrombin

vibrant-m

SAE0110

HRV-3C Protease, Biotin tagged

storage temp.

−20°C

storage temp.

−70°C

storage temp.

-

storage temp.

−20°C

recombinant

expressed in HEK 293 cells

recombinant

expressed in HEK 293 cells

recombinant

-

recombinant

expressed in E. coli

concentration

≥5000 units/mL

concentration

-

concentration

-

concentration

0.8-1.2 mg/mL

General description

Thrombin is an endolytic serine protease that selectively cleaves the Arg–Gly bonds of fibrinogen to form fibrin and release fibrinopeptides A and B.1,2
The optimal cleavage sites for thrombin are as follows:
1. A-B-Pro-Arg-||-X-Y, where A and B are hydrophobic amino acids, and X and Y are nonacidic amino acids.
2. Gly-Arg-||-Gly
Recombinant human thrombin protease is expressed in human HEK 293 cells as a glycoprotein heterodimer. The DTT-reduced protein migrates as two bands of ∼31 kDa (heavy chain) and ∼6 kDa (light chain) on SDS-PAGE. This protein is manufactured in human cells, with no serum. The human cells expression system allows human-like glycosylation and folding, and often supports higher activity of the protein.
This thrombin protease is useful for cleaving recombinant proteins that are expressed as fusion proteins with this sequence between the carrier domain and the protein of interest.

Application

This biotinylated thrombin protease can be used for on-column cleavage of fusion proteins with a thrombin cleavage site. It specifically cleaves the protein of interest from a column-bound fusion protein, leaving the fusion domain or tag bound to the affinity column (e.g., Ni-NTA column) and eluting only the protein of interest.
This method is advantageous over post-elution cleavage for several reasons:
  • It eliminates most impurities normally associated with purification on Ni-chelating columns.
  • It allows gentler elution conditions, with added flexibility in the elution buffer composition. This can mitigate protein aggregation and inactivation.
After cleavage, the protease can be removed with any avidin-conjugated or streptavidin-conjugated beads.

Features and Benefits

The product is supplied in an aqueous buffer of pH 6.0, with 50% (v/v) glycerol.
Catalytic pH range:11.5–10
Optimal pH:11-8.3
(Note: thrombin precipitates at pH ≤5)
Molecular mass:4,12 37.4 kDa
Human isozymes pI range:6.35–7.6
E280 (1%):12 -18.3
This Thrombin protease, biotin-tagged enables on-column cleavage of fusion proteins with a thrombin cleavage site.

Storage Class

10 - Combustible liquids

wgk_germany

WGK 1


Certificates of Analysis (COA)

Search for Certificates of Analysis (COA) by entering the products Lot/Batch Number. Lot and Batch Numbers can be found on a product’s label following the words ‘Lot’ or ‘Batch’.

Already Own This Product?

Documents related to the products that you have purchased in the past have been gathered in the Document Library for your convenience.

Visit the Document Library

Difficulty Finding Your Product Or Lot/Batch Number?

Product numbers are combined with Pack Sizes/Quantity when displayed on the website (example: T1503-25G). Please make sure you enter ONLY the product number in the Product Number field (example: T1503).

Example:

T1503
Product Number
-
25G
Pack Size/Quantity

Additional examples:

705578-5MG-PW

PL860-CGA/SHF-1EA

MMYOMAG-74K-13

1000309185

enter as 1.000309185)

Having trouble? Feel free to contact Technical Service for assistance.

Lot and Batch Numbers can be found on a product's label following the words 'Lot' or 'Batch'.

Aldrich Products

  • For a lot number such as TO09019TO, enter it as 09019TO (without the first two letters 'TO').

  • For a lot number with a filling-code such as 05427ES-021, enter it as 05427ES (without the filling-code '-021').

  • For a lot number with a filling-code such as STBB0728K9, enter it as STBB0728 without the filling-code 'K9'.

Not Finding What You Are Looking For?

In some cases, a COA may not be available online. If your search was unable to find the COA you can request one.

Request COA

Articles

Proteases for biotinylated tag removal for protein purification workflows with related reagents and technical resources.

Our team of scientists has experience in all areas of research including Life Science, Material Science, Chemical Synthesis, Chromatography, Analytical and many others.

Contact Technical Service