FK506 binding protein, 12 kilodalton molecular weight (FKBP12), also called FKBP1, was originally characterized as a peptidyl-prolyl isomerase that catalyzes the transition between cis- and trans-proline residues critical for proper folding of proteins. The macrolide immunosuppressants FK506 (Tacrolimus) and rapamycin bind to FKBP12 with high affinity, while the structurally related compound cyclosporine binds with a much lower affinity (1). The binding of these drugs causes FKBP12 to become a potent inhibitor of calcineurin phosphatase activity (2) and TOR kinase activity (3). The inhibition of protein phosphatase activity is highly selective for calcineurin (2), making the FK506/FKBP12 complex a useful tool in the study of this enzyme. Knockout mice lacking FKBP12 are morphologically normal, but develop cardiomyopathies that may be related to dysregulation of ryanodyne receptors (4).
Recombinant Human FKBP12 Protein, CF
R&D Systems | Catalog # 3777-FK
Loading...
Key Product Details
- R&D Systems E. coli-derived Recombinant Human FKBP12 Protein (3777-FK)
- 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
Applications
Inhibition Activity
Loading...
Product Specifications
Source
E. coli-derived human FKBP12 protein
Gly2-Glu108, with an N-terminal Met and 6-His tag
Gly2-Glu108, with an N-terminal Met and 6-His tag
Purity
>95%, by SDS-PAGE under reducing conditions and visualized by silver stain.
Endotoxin Level
<1.0 EU per 1 μg of the protein by the LAL method.
N-terminal Sequence Analysis
Met
Predicted Molecular Mass
13 kDa
SDS-PAGE
13 kDa, reducing conditions
Activity
Measured by its ability to convert the substrate, Suc-AAPF-pNA, from Cis to Trans formation.
The specific activity is >2500 pmol/min/μg, as measured under the described conditions.
The specific activity is >2500 pmol/min/μg, as measured under the described conditions.
Formulation, Preparation, and Storage
3777-FK
| Formulation | Supplied as a 0.2 μm filtered solution in HEPES and NaCl. |
| Shipping | The product is shipped with dry ice or equivalent. Upon receipt, store it immediately at the temperature recommended below. |
| Stability & Storage | Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
|
Background: FKBP12
References
- Hamilton, G.S. and J.P. Steiner (1998) J. Med. Chem. 41:5119.
- Liu, J. et al. (1992) Biochemistry 31:3896.
- Toral-Barza, L. et al. (2005) Biochem. Biophys. Res. Comm. 332:304.
- Hamilton, S.L and M.M. Matzuk (1998) Nature 391:489.
Long Name
12 kDa FK506 Binding Protein
Alternate Names
FKBP1a, PKC12, PPIASE
Gene Symbol
FKBP1A
UniProt
Additional FKBP12 Products
Product Documents for Recombinant Human FKBP12 Protein, CF
Certificate of Analysis
To download a Certificate of Analysis, please enter a lot or batch number in the search box below.
Note: Certificate of Analysis not available for kit components.
Product Specific Notices for Recombinant Human FKBP12 Protein, CF
For research use only
Related Research Areas
Citations for Recombinant Human FKBP12 Protein, CF
Powered by Bioz
Customer Reviews for Recombinant Human FKBP12 Protein, CF
There are currently no reviews for this product. Be the first to review Recombinant Human FKBP12 Protein, CF and earn rewards!
Have you used Recombinant Human FKBP12 Protein, CF?
Submit a review and receive an Amazon gift card!
$25/€18/£15/$25CAN/¥2500 Yen for a review with an image
$10/€7/£6/$10CAN/¥1110 Yen for a review without an image
Submit a review
Protocols
View specific protocols for Recombinant Human FKBP12 Protein, CF (3777-FK):
Materials
*Adjusted for Blank
**Using the extinction coefficient 9300 M-1cm-1
***Using the path length 1 cm
Note: the output of many spectrophotometers is in mOD. Per Reaction:
- Assay Buffer: 50 mM HEPES, 100 mM NaCl, pH 8.0
- Recombinant Human FKBP12 (rhFKBP12 ) (Catalog # 3777-FK)
- Substrate: Suc-AAPF-pNA (Sigma, Catalog # S7388)
- Chymotrypsin (Sigma, Catalog # C3142), 60 mg/mL stock in 1 mM HCl
- Lithium Chloride (Sigma, Catalog # 213233)
- Trifluoroethanol (Fisher, Catalog # BP622)
- Ethanol
- Quartz Micro-Cuvette (VWR, Catalog # 414004-050) or equivalent
- Plate Reader with Cuvette Port (Model: SpectraMax Plus by Molecular Devices) or equivalent
Note: Time and temperature are critical to this assay. Execute substrate addition and reading steps as quickly as possible, and keep components on ice while not in use.
- Prepare Assay Buffer and place on ice.
- Prepare 20 mg/mL Lithium Chloride in Trifluoroethanol.
- Prepare 3 mM Suc-AAPF-pNA in 20 mg/mL Lithium Chloride (step 2) and place on ice. Use 3 mM Suc-AAPF-pNA within 4 hours of preparation.
- Combine 170 µL of cold Assay Buffer and 20 µL of cold 60 mg/mL Chymotrypsin in a clean micro-cuvette to create a Blank.
- Place micro-cuvette on ice for 10 minutes.
- Quickly add 10 µL of cold 3 mM Suc-AAPF-pNA. Invert vigorously 1-2 seconds and wipe condensation off micro-cuvette.
- Read immediately at an absorbance of 405 nm for 1 minute in kinetic mode.
- Clean micro-cuvette thoroughly with ethanol and cold deionized water.
- Dilute rhFKBP12 to 100 µg/mL in cold Assay Buffer.
- Combine 160 µL of cold Assay Buffer, 20 µL of cold 60 mg/mL Chymotrypsin, and 10 µL of cold 100 µg/mL rhFKBP12 in a clean micro-cuvette.
- Place micro-cuvette on ice for 10 minutes.
- Quickly add 10 µL of cold 3 mM Suc-AAPF-pNA. Invert vigorously 1-2 seconds and wipe condensation off micro-cuvette.
- Read immediately at an absorbance of 405 nm for 1 minute in kinetic mode.
- Clean micro-cuvette thoroughly with ethanol and cold deionized water.
- Use the first 30 seconds of the reads to determine the test rate.
|
Specific Activity (pmol/min/µg) = |
Adjusted Rate* (OD/min) x well volume (L) x 1012 pmol/mol |
| ext. coeff** (M-1cm-1) x path length*** (cm) x amount of enzyme (µg) |
*Adjusted for Blank
**Using the extinction coefficient 9300 M-1cm-1
***Using the path length 1 cm
Note: the output of many spectrophotometers is in mOD. Per Reaction:
- rhFKBP12: 1 µg
- Suc-AAPF-pNA: 150 µM
- Chymotrypsin: 1.2 mg