Recombinant Variola CRMB Protein

Carrier Free

Catalog # Availability Size / Price Qty
4778-CR-010/CF

With Carrier

Catalog # Availability Size / Price Qty
4778-CR-010
R&D Systems Recombinant Proteins and Enzymes
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Recombinant Variola CRMB Protein Summary

Product Specifications

Purity
>95%, by SDS-PAGE under reducing conditions and visualized by silver stain
Endotoxin Level
<0.01 EU per 1 μg of the protein by the LAL method.
Activity
Measured by its ability to inhibit the TNF-alpha mediated cytotoxicity in the L‑929 mouse fibroblast cells in the presence of the metabolic inhibitor actinomycin D. Gileva, I.P. et al. (2006) Biochim. Biophys. Acta. 1764:1710. The ED50 for this effect, in the presence of 0.25 ng/mL of rhTNF-alpha, is
0.6-2.4 ng/mL.
Source
Mouse myeloma cell line, NS0-derived viral CRMB protein
Ala23-Leu349, with a C-terminal 6-His tag
Accession #
N-terminal Sequence
Analysis
Ala23
Structure / Form
Disulfide-linked homodimer
Predicted Molecular Mass
36.5 kDa (monomer)
SDS-PAGE
55 - 60 kDa, reducing conditions

Product Datasheets

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4778-CR (with carrier)

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4778-CR/CF (carrier free)

Carrier Free

What does CF mean?

CF stands for Carrier Free (CF). We typically add Bovine Serum Albumin (BSA) as a carrier protein to our recombinant proteins. Adding a carrier protein enhances protein stability, increases shelf-life, and allows the recombinant protein to be stored at a more dilute concentration. The carrier free version does not contain BSA.

What formulation is right for me?

In general, we advise purchasing the recombinant protein with BSA for use in cell or tissue culture, or as an ELISA standard. In contrast, the carrier free protein is recommended for applications, in which the presence of BSA could interfere.

4778-CR

Formulation Lyophilized from a 0.2 μm filtered solution in PBS with BSA as a carrier protein.
Reconstitution Reconstitute at 100 μg/mL in sterile PBS containing at least 0.1% human or bovine serum albumin.
Shipping The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below.
Stability & Storage: Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
  • 12 months from date of receipt, -20 to -70 °C as supplied.
  • 1 month, 2 to 8 °C under sterile conditions after reconstitution.
  • 3 months, -20 to -70 °C under sterile conditions after reconstitution.

4778-CR/CF

Formulation Lyophilized from a 0.2 μm filtered solution in PBS
Reconstitution Reconstitute at 100 μg/mL in sterile PBS.
Shipping The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below.
Stability & Storage: Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
  • 12 months from date of receipt, -20 to -70 °C as supplied.
  • 1 month, 2 to 8 °C under sterile conditions after reconstitution.
  • 3 months, -20 to -70 °C under sterile conditions after reconstitution.
Reconstitution Calculator

Reconstitution Calculator

The reconstitution calculator allows you to quickly calculate the volume of a reagent to reconstitute your vial. Simply enter the mass of reagent and the target concentration and the calculator will determine the rest.

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Background: CRMB

Variola virus is the causative agent of human smallpox. Like other poxviruses, it encodes a variety of molecules that shield virus infected cells from immune clearance. The cytokine response modifiers CRMB, C, D, and E, which are differentially expressed among the poxviruses, function as decoy TNF receptors and block the proinflammtatory and antiviral effects of TNF (1, 2). Of the CRM proteins, Variola virus encodes only CRMB, secreted from virus infected cells as a 90 kDa disulfide linked dimer (3). The N-terminal 112 amino acid (aa) region of CRMB mediates binding to human, mouse, and rat TNF as well as human lymphotoxin-alpha, and neutralizes the cytolytic effects of TNF (3, 4). The C-terminal 155 aa region of CRMB, known as a SECRET domain (smallpox virus-encoded chemokine receptor), binds the chemokines CCL25, CCL28, CXCL12b, CXCL13, and CXCL14, which are involved in the antiviral immune response (4). Functionally, the SECRET domain interferes with the in vitro migration of T cells in response to CCL25 (4). A SECRET domain is also present in CRMD but not in CRMC or CRME. Variola virus CRMB shares 84% - 92% aa sequence identity with camelpox virus, cowpox virus, and monkeypox virus CRMB, but only 21% with vaccinia virus CRMB (which lacks a SECRET domain). The TNF binding domain of CRMB shares 30% and 42% aa sequence identity with comparable regions of human TNF R1 and R2, respectively.

References
  1. Johnston, J.B. and G. McFadden (2003) J. Virol. 77:6093.
  2. Massung, R.F. et al. (1993) Nature 366:748.
  3. Gileva, I.P. et al. (2006) Biochim. Biophys. Acta 1764:1710.
  4. Alejo, A. et al. (2006) Proc. Natl. Acad. Sci. 103:5995.
Long Name
Cytokine Response-modifying Protein B
Alternate Names
CRMB; G2R; G4R

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