Home » XIAP » XIAP Proteins » Recombinant Human XIAP (BIR3) Protein, CF
Novus Biologicals products are now on bio-techne.com

Recombinant Human XIAP (BIR3) Protein, CF

 (2 Publications)
 See All Related
Submit a Review

Images

 
There are currently no images for XIAP (895-XB).

Every product we sell is backed by Novus' 100% Guarantee. If you have used this product, please submit your images and reviews to earn reward points.

Product Details

Summary
Reactivity HuSpecies Glossary
Applications Inhibition Activity
Format
Carrier-Free

Order Details

Novus Biologicals is part of Bio-Techne

Shop this product on bio-techne.com

Novus Biologicals, part of Bio-Techne, offers 100% Guarantee

Recombinant Human XIAP (BIR3) Protein, CF Summary

Details of Functionality
Measured by its ability to inhibit DEVD-AFC cleavage activity in cell extracts activated by addition of cytochrome c and dATP. The IC50 for this effect is <300 nM.
Optimal dilutions should be determined by each laboratory for each application.
Source
E. coli-derived human XIAP protein
Asn252-Thr356, with an N-terminal Met and 6-His tag
Accession #
N-terminal Sequence
Met
Protein/Peptide Type
Recombinant Enzymes
Gene
XIAP
Purity
>95%, by SDS-PAGE under reducing conditions and visualized by silver stain

Applications/Dilutions

Dilutions
  • Inhibition Activity
Theoretical MW
13 kDa.
Disclaimer note: The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors.
SDS-PAGE
11 kDa, reducing conditions
Publications
Read Publications using
895-XB in the following applications:

Packaging, Storage & Formulations

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.
Buffer
Supplied as a 0.2 μm filtered solution in HEPES, NaCl and DTT.
Purity
>95%, by SDS-PAGE under reducing conditions and visualized by silver stain
Assay Procedure
  • Jurkat E6 wild type cell extracts (see supplementary methods for preparation)
  • Extraction Buffer: 50 mM HEPES, 10 mM KCl, 5 mM EGTA, 1 mM MgCl2, 0.2% CHAPS, 0.2 mM DTT, pH 7.5
  • Assay Buffer: 10 mM HEPES, 0.5 mM EGTA, 5 mM DTT, 10% Glycerol, pH 7.5
  • Formulation Buffer: 25 mM HEPES, 0.1 M NaCl, 1 mM DTT, pH 8.5
  • Recombinant Human XIAP BIR3 Domain  (rhXIAP) (BIR3) (Catalog # 895-XB)
  • Cytochrome C, Bovine heart (Sigma, Catalog # C3131), 2 mg/mL stock in deionized water
  • dATP (Sigma, Catalog # D6500), 10 mM stock adjusted to pH 7.5 with NaOH
  • Substrate: Ac-Asp-Glu-Val-Asp-AFC (DEVD-AFC) (MP Biomedicals, Catalog # AFC138), 10 mM stock in DMSO
  • F16 Black Maxisorp Plate (Nunc, Catalog # 475515)
  • Fluorescent Plate Reader (Model: SpectraMax Gemini EM by Molecular Devices) or equivalent
Note: All reagents and assay components should be kept on ice until use.
  1. Thaw cell extracts and centrifuge in a microcentrifuge at 14,000 rpms for 5 minutes at 4 °C. Transfer supernatants to chilled tubes and use within 1 hour.
  2. Prepare a curve of rhXIAP (BIR3) MW: 13,047 Da) in Formulation Buffer. Make the following serial dilutions: 5000, 2500, 1500, 500, 250, 50, 25, and 5 nM. Note: High point may not be achievable depending on lot received.
  3. Prepare the activator mixture by combining equal volumes of 2 mg/mL Cytochrome C and 10 mM dATP for working concentrations of 1 mg/mL and 5 mM, respectively.
  4. Prepare reaction mixtures in tubes by combining 10 μL of each rhXIAP (BIR3)  curve dilution, 10 μL of cell extract supernatant, and 5 μL of the cytochrome C/dATP activator mixture. Also, prepare the following controls:
    1. Total Control: 10 μL of Extraction Buffer, 10 μL of cell extract supernatant, and 5 μL of the cytochrome C/dATP activator mixture.
    2. Inactive Control: 15 μL of Extraction Buffer and 10 μL of cell extract supernatant.  The total reaction volume is 25 μL.
  5. Incubate for 60 minutes at 37 °C.
  6. After incubation, add 100 μL of Assay Buffer to each vial for a five-fold dilution. Mix briefly.
  7. Dilute Substrate to 100 μM in Assay Buffer.
  8. In a plate load 50 μL of diluted incubated reaction mixtures, and start the reaction by adding 50 μL of 100 μM Substrate.
  9. Read at excitation and emission wavelengths of 400 nm and 505 nm, respectively, in kinetic mode for 5 minutes.
  10. Derive the 50% inhibiting concentration (IC50) of rhXIAP (BIR3)  by plotting normalized activity vs. reaction concentration of rhXIAP (BIR3) (step 4) with 4-PL fitting.
  11. Normalized activity may be determined using the following equation:
         % Normalized Activity = Sample (RFU/min) - Inactive Control** (RFU/min) x 100%
    Total Control (RFU/min)
Per Reaction:
  • rhXIAP (BIR3) curve:  2000, 1000, 600, 200, 100, 20, 10, and 2 nM
  • Substrate: 50 μM

Notes

This product is produced by and ships from R&D Systems, Inc., a Bio-Techne brand.

Alternate Names for Recombinant Human XIAP (BIR3) Protein, CF

  • API3X-linked inhibitor of apoptosis protein
  • apoptosis inhibitor 3
  • baculoviral IAP repeat-containing 4
  • baculoviral IAP repeat-containing protein 4
  • BIRC4
  • BIRC4XLP2
  • E3 ubiquitin-protein ligase XIAP
  • EC 6.3.2.-
  • hIAP3
  • hIAP-3
  • hILP
  • IAP3
  • IAP-3
  • IAP-like protein
  • ILP
  • ILP1
  • Inhibitor of apoptosis protein 3
  • MIHA
  • XIAP
  • X-linked IAP
  • X-linked inhibitor of apoptosis

Background

XIAP (X-chromosome linked inhibitor of apoptosis) is a member of the inhibitor of apoptosis (IAP) family of proteins that inhibit caspases. The BIR3 domain inhibits caspase-9. The ability of XIAP (BIR3 domain) to inhibit caspases is prevented by SMAC/Diablo.

  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.

Customers Who Viewed This Item Also Viewed...

Caspase-3 Antibody
[Unconjugated] - Acti ...
AF835
Immunocytochemistry Caspase-3 Antibody [Unconjugated] - Active
Immunohistochemistry Caspase-3 Antibody [Unconjugated] - Active
Immunocytochemistry Caspase-3 Antibody [Unconjugated] - Active
(7 Reviews)
(387 Publications)
Species: Hu, Mu
Applications: ICC, IHC
SMAC/Diablo
Antibody
NB100-56311
Western Blot SMAC/Diablo Antibody
Immunohistochemistry SMAC/Diablo Antibody
(16 Publications)
Species: Hu, Mu
Applications: ICC/IF, IHC, IHC-P, WB
Bcl-xL Antibody
NB100-56104
Western Blot Bcl-xL Antibody
Immunohistochemistry-Paraffin Bcl-xL Antibody
Western Blot Bcl-xL Antibody
(1 Review)
(3 Publications)
Species: Ca, Hu, Mu, Rt
Applications: IHC, IHC-Fr, IHC-P, IP, WB
Bcl-2 Antibody
NB100-56098
Western Blot Bcl-2 Antibody
Western Blot Bcl-2 Antibody
Western Blot Bcl-2 Antibody
(2 Reviews)
(26 Publications)
Species: Ca, Hu, Mu, Rt
Applications: IHC, IHC-P, IP, WB
TNF-alpha
[Unconjugated]
210-TA
SDS-PAGE TNF-alpha [Unconjugated]
Bioactivity TNF-alpha [Unconjugated]
SEC-MALS TNF-alpha [Unconjugated]
(3 Reviews)
(811 Publications)
Species: Hu
Applications: BA
TRAIL/TNFSF10
[Unconjugated]
375-TL
TRAIL/TNFSF10 [Unconjugated]
(1 Review)
(54 Publications)
Species: Hu
Applications: BA
Caspase-9 Antibody
NB100-56118
Immunohistochemistry Caspase-9 Antibody
Immunohistochemistry Caspase-9 Antibody
Immunohistochemistry Caspase-9 Antibody
(1 Review)
(28 Publications)
Species: Ca, Ma, Hu, Mu, Rt
Applications: Flow-IC, ICC/IF, IHC, IHC-Fr, IHC-P, IP, WB
Survivin Antibody
- BSA Free
NB500-201
Flow Cytometry Survivin Antibody - BSA Free
Dual RNAscope ISH-IHC Survivin Antibody - BSA Free
Knockdown Validated Survivin Antibody - BSA Free
(22 Reviews)
(374 Publications)
Species: Ca, Fe, Gp, Ha, Hu, Mu, Rt
Applications: ChIP, Dual ISH-IHC, ELISA, Flow, ICC/IF, IHC, IHC-Fr, IHC-P, IP, KD, Simple Western, WB
Amylin Antibody
NBP2-33680
Immunohistochemistry Amylin Antibody
Immunohistochemistry-Paraffin Amylin Antibody
Immunohistochemistry-Paraffin Amylin Antibody
Species: Hu
Applications: IHC, IHC-P
Alkaline
Phosphatase, Intestin ...
NBP3-25355
Western Blot Alkaline Phosphatase, Intestinal Antibody (HL1268) - Azide and BSA Free
Immunohistochemistry-Paraffin Alkaline Phosphatase, Intestinal Antibody (HL1268) - Azide and BSA Free
Western Blot Alkaline Phosphatase, Intestinal Antibody (HL1268) - Azide and BSA Free
Species: Dr, Hu, Mu
Applications: IHC, IHC-P, WB
CD47 Antibody
[Unconjugated]
AF4670
Western Blot CD47 Antibody [Unconjugated]
Neutralization CD47 Antibody [Unconjugated]
Immunohistochemistry CD47 Antibody [Unconjugated]
(4 Reviews)
(14 Publications)
Species: Hu
Applications: CyTOF-ready, Flow, IHC, KO, Neut, WB
MAGT1 Antibody
NBP2-14214
Immunohistochemistry-Paraffin MAGT1 Antibody
Immunohistochemistry-Paraffin MAGT1 Antibody
Immunohistochemistry-Paraffin MAGT1 Antibody
Species: Hu
Applications: IHC, IHC-P
Caspase-8 Antibody
NB100-56116
Western Blot Caspase-8 Antibody
Immunohistochemistry-Paraffin Caspase-8 Antibody
Immunohistochemistry-Paraffin Caspase-8 Antibody
(2 Reviews)
(79 Publications)
Species: Ma, Hu, Mu, Rt
Applications: Flow, ICC/IF, IHC, IHC-Fr, IHC-P, IP, Simple Western, WB
cIAP-1/HIAP-2
Antibody [Unconjugate
AF8181
Western Blot cIAP-1/HIAP-2 Antibody [Unconjugated]
Immunohistochemistry cIAP-1/HIAP-2 Antibody [Unconjugated]
Knockout Validated cIAP-1/HIAP-2 Antibody [Unconjugated]
(4 Reviews)
(66 Publications)
Species: Hu
Applications: IHC, KO, Simple Western, WB
Cytochrome c
Antibody (7H8.2C12) - ...
NB100-56503
Flow Cytometry Cytochrome c Antibody (7H8.2C12) - BSA Free
Flow (Intracellular) Cytochrome c Antibody (7H8.2C12) - BSA Free
Western Blot Cytochrome c Antibody (7H8.2C12) - BSA Free
(61 Publications)
Species: Dr, Hu, Mu, Rb, Rt
Applications: Flow-CS, Flow-IC, Flow, IB, ICC/IF, IHC, IHC-P, Simple Western, WB
NFkB p105/p50
Antibody (2J10D7) - B ...
NB100-56583
Flow Cytometry NFkB p105/p50 Antibody (2J10D7) - BSA Free
Flow Cytometry NFkB p105/p50 Antibody (2J10D7) - BSA Free
Flow Cytometry NFkB p105/p50 Antibody (2J10D7) - BSA Free
(1 Review)
(5 Publications)
Species: Hu, Rb, Rt
Applications: Flow, IHC, IHC-P, KO, WB
AKT [p Ser473]
Antibody [Unconjugate ...
AF887
Western Blot AKT [p Ser473] Antibody [Unconjugated] - Pan Specific
Simple Western AKT [p Ser473] Antibody [Unconjugated] - Pan Specific
Intracellular Staining by Flow Cytometry AKT [p Ser473] Antibody [Unconjugated] - Pan Specific
(49 Publications)
Species: Hu, Mu, Rt
Applications: CyTOF-ready, IHC, ICFlow, Simple Western, WB
Bax Antibody (6A7)
NBP1-28566
Western Blot Bax Antibody (6A7)
Western Blot Bax Antibody (6A7)
Immunohistochemistry-Frozen Bax Antibody (6A7)
(3 Reviews)
(20 Publications)
Species: Bv, Hu, Pm, Mu, Po, Rt
Applications: Func, IHC, IHC-Fr, IHC-P, WB

Publications for XIAP (895-XB)(2)

We have publications tested in 1 confirmed species: Human.

We have publications tested in 2 applications: Bioassay, Western Blot.


Filter By Application
Bioassay
(1)
Western Blot
(1)
All Applications
Filter By Species
Human
(2)
All Species

Reviews for XIAP (895-XB) (0)

There are no reviews for XIAP (895-XB). By submitting a review you will receive an Amazon e-Gift Card or Novus Product Discount.
  • Review with no image -- $10/€7/£6/$10 CAD/¥70 Yuan/¥1110 Yen
  • Review with an image -- $25/€18/£15/$25 CAD/¥150 Yuan/¥2500 Yen

FAQs for XIAP (895-XB) (0)

There are no specific FAQs related to this product. Read our general customer & technical service FAQs.

Additional XIAP Products

Blogs on XIAP.

Check out the latest blog posts on XIAP.
The Ins and Outs of Survivin
By Rachel M.A. Linger, Ph.D.What is survivin?Survivin is a small (16.5 kDa) protein normally found in human fetal tissue. In contrast, survivin is typically undetectable in most normal adult tissues. Expression of ...  Read full blog post.
Apoptosis and Necroptosis Part II: Inhibitors of apoptosis proteins (IAPs); Key regulators of the balance between necroptosis, apoptosis and survival
In the first installment of this two-part blog post titled "Apoptosis and Necroptosis: Important factors to identify both types of programmed cell death", the mechanisms by which cell death occurs and ways to identify these pathways were ...  Read full blog post.
Caspase 9 - an important apoptosis marker
Caspases are essential mediators of programmed cell death and are needed for both the induction of apoptosis as well as for aiding the degradation of cellular structures. Initiator caspases (such as Caspase-9) sense and respond to various signals i...  Read full blog post.
cIAP2 - balancing cell death and cell survival
The inhibitor of apoptosis proteins (IAPs) are important regulators of cell death and inflammation. The cellular inhibitor of apoptosis protein 2 (cIAP2) contains three Baculovirus IAP repeat (BIR) domains, a Ubiquitin associated (UBA) domain, and ...  Read full blog post.
Read our latest blog and use the new citation tool on bio-techne.com

Customers Who Bought This Also Bought

Caspase&#x2011;3 was detected in immersion fixed anti-FAS treated Jurkat human acute T cell leukemia cell line using 0.3 &micro;g/mL Human/Mouse Active Caspase&#x2011;3 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF835) for 3 hours at room temperature. Cells were stained (red) and counterstained (green). View our protocol for Fluorescent ICC Staining of Cells on Coverslips.
Caspase-3 Antibody [Unconjugated] - Active
AF835
1 µg/lane of Recombinant Human TNF-alpha was resolved by SDS-PAGE with silver staining, under reducing (R) conditions, showing a band at 17 kDa.
TNF-alpha [Unconjugated]
210-TA-005

Contact Information

Product PDFs

  PDF SDS
  PDF Datasheet
  Request CofA

Calculators

Concentration Calculator

The concentration calculator allows you to quickly calculate the volume, mass or concentration of your vial. Simply enter your mass, volume, or concentration values for your reagent and the calculator will determine the rest.

=
÷

Review this Product

Be the first to review our Recombinant Human XIAP (BIR3) Protein, CF and receive a gift card or discount.


Bioinformatics

Gene Symbol XIAP
Uniprot