Recombinant Human XIAP (BIR2) Protein, CF

R&D Systems | Catalog # 786-XB

R&D Systems
100% Guarantee

Key Product Details

  • R&D Systems E. coli-derived Recombinant Human XIAP (BIR2) Protein (786-XB)
  • 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

U45880

Applications

Inhibition Activity
View all XIAP Proteins and Enzymes »

Product Specifications

Source

E. coli-derived human XIAP protein
Arg124-Asp242, with an N-terminal Met and 6-His 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

15 kDa

SDS-PAGE

12 kDa, reducing conditions

Activity

Measured by its ability to inhibit DEVD-AFC cleavage activity by Recombinant Human Caspase‑7 (Catalog # 823-C7).
The IC50 for inhibition of Recombinant Human (rh) Caspase-7 (Catalog # 823-C7) activity is 2-20 nM, as measured under the described conditions.

Formulation, Preparation, and Storage

786-XB
Formulation Supplied as a 0.2 μm filtered solution in HEPES, NaCl, DTT and Glycerol.
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.
  • 6 months from date of receipt, -20 to -70 °C as supplied.
  • 3 months, -20 to -70 °C under sterile conditions after opening.

Background: XIAP

XIAP (X-chromosome linked inhibitor of apoptosis) is a member of the inhibitor of apoptosis (IAP) family of proteins that inhibit caspases. The BIR2 domain of XIAP inhibits Caspase-3 and Caspase-7. The ability of XIAP to inhibit caspases is prevented by SMAC/Diablo through binding to XIAP-BIR2 and -BIR3 domains.

 

SUPPLEMENTARY METHODS 

 

XIAP (BIR2 domain) inhibits DEVD-AFC cleavage by activated cell extracts.

 

XIAP (BIR2 domain) inhibits DEVD-AFC cleavage by Recombinant Human Caspase-3

Recombinant Human Caspase‑3 (Catalog # http://www.rndsystems.com/product_results.aspx?k=707-C3">707-C3) can be used in the Activity Assay Protocol in place of Casapase-7 by diluting to initial concentration of 32 pg/uL in Step 2.

The IC50 for inhibition of rhCaspase-3 activity is typically 10-100 nM.

 

References

  1. Deveraux, Q. et al. (1998) EMBO J. 17(8):2215.
  2. Deveraux, Q. et al. (1999) EMBO J. 18(19):5242.
  3. Verhagen, A. et al. (2000) Cell 102:43.
  4. Chai, J. et al. (2001) Cell 104:769.
  5. Riedl, S. (2001) Cell 104:791.
  6. Suzuki, Y. (2001) J. Biol. Chem. 276(29):27058.
  7. Ekert, P. et al. (2001) J. Cell Biol. 152(3):483.
  8. Du, C. et al. (2000) Cell 102:33.

Long Name

X-linked Inhibitor of Apoptosis

Alternate Names

BIRC4

Entrez Gene IDs

331 (Human); 11798 (Mouse); 63879 (Rat)

Gene Symbol

XIAP

UniProt

U45880 (Human)

Additional XIAP Products

Product Documents for Recombinant Human XIAP (BIR2) 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.

Download SDS

Product Specific Notices for Recombinant Human XIAP (BIR2) Protein, CF

For research use only

Related Research Areas

Cysteine Proteases and Regulators

Customer Reviews for Recombinant Human XIAP (BIR2) Protein, CF

There are currently no reviews for this product. Be the first to review Recombinant Human XIAP (BIR2) Protein, CF and earn rewards!

Have you used Recombinant Human XIAP (BIR2) 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
Amazon Gift Card

Protocols

View specific protocols for Recombinant Human XIAP (BIR2) Protein, CF (786-XB):

Materials
  • Activation Buffer: 50 mM HEPES, 100 mM NaCl, 1 mM EDTA, 5 mM DTT, 0.1% CHAPS, 10% Sucrose, pH 7.5
  • Assay Buffer: 25 mM HEPES, 0.1 M NaCl, 1 mM DTT, pH 8.0
  • F16 Black Maxisorp Plate (Nunc, Catalog # 475515)
  • Recombinant Human Caspase‑7 (rhCaspase‑7) (Catalog # 823-C7)
  • Recombinant Human XIAP BIR2 Domain (rhXIAP-BIR2) (Catalog # 786-XB)
  • Substrate: Ac-Asp-Glu-Val-Asp-AFC (MP Biomedicals, Catalog # AFC138), 10 mM stock in DMSO
  1. Dilute rhXIAP-BIR2 (MW: 14,801Da) to various concentrations in Assay Buffer. Make serial dilutions of: 8000, 2000, 1000, 300, 100, 50, 30, 10, 3, and 1 nM.
  2. Dilute rhCaspase-7 to 0.12 ng/µL in Activation Buffer.
  3. Load Plate in the following order:
    70 µL Assay Buffer
    10 µL of the rhXIAP-BIR2 dilutions
    10 µL of 0.12 ng/µL rhCaspase-7
  4. Include a control, in duplicate, containing Assay Buffer and rhCaspase-7 only.
  5. Incubate at RT for 15 minutes.
  6. Dilute Substrate in Assay Buffer to 500 µM.
  7. Load 10 µL Substrate per well to start the reaction.
  8. Read at excitation and emission wavelengths of 400 nm and 505 nm, respectively in kinetic mode for 5 minutes.

  9. Derive the 50% inhibiting concentration (IC50) for rhXIAP-BIR2 by plotting RFU/min (or specific activity) vs. concentration with 4-PL fitting.

  10. The specific activity for rhCaspase-7 at each point may be determined using the following formula (if needed):

     Specific Activity (pmol/min/µg) =

 Adjusted Vmax* (RFU/min) x Conversion Factor** (pmol/RFU)
amount of enzyme (µg)

     *Adjusted for Substrate Blank
     **Derived using calibration standard 7-amino, 4-trifluoromethylcoumarin (AFC) (Calbiochem, Catalog # 164580).

Per Well:

  • rhCaspase-7: 0.0012 μg
  • rhXIAP-BIR2 curve: 800, 200, 100, 30, 10, 5, 3, 1, 0.3 and 0.1 nM
  • Substrate: 50 µM

FAQs

No product specific FAQs exist for this product.

View all FAQs for Proteins and Enzymes