Recombinant Human IL-15R alpha Fc Chimera Avi-tag, CF

Catalog #: AVI7194 Datasheet / COA / SDS

Biotinylated

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AVI7194-050

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Summary Product Datasheets Carrier Free Data Images Reconstitution Calculator Background Related Research Areas

Recombinant Human IL-15R alpha Fc Chimera Avi-tag, CF Summary

Learn more about Avi-tag Biotinylated Proteins

Product Specifications

Purity

>95%, by SDS-PAGE visualized with Silver Staining and quantitative densitometry by Coomassie® Blue Staining.

Endotoxin Level

<0.10 EU per 1 μg of the protein by the LAL method.

Activity

The biotin to protein ratio is greater than 0.7 as determined by the HABA assay. Measured by its binding ability in a functional ELISA. When Biotinylated Recombinant Human IL-15R alpha Fc Chimera Avi-tag (Catalog # AVI7194) is immobilized at 0.5 µg/mL (100 µL/well), Recombinant Human IL-15 (Catalog # 247-ILB) binds with an ED₅₀ of 0.02-0.16 ng/mL.

Source

Chinese Hamster Ovary cell line, CHO-derived human IL-15R alpha protein

Human IL-15R alpha (Ile31-Thr205) Accession # Q13261.1	IEGRMD	Human IgG₁ (Pro100-Lys330)	Avi-tag
N-terminus			C-terminus

Accession #

Q13261.1

N-terminal Sequence
Analysis

Ile31

Structure / Form

Disulfide-linked homodimer, biotinylated via Avi-tag

Predicted Molecular Mass

47 kDa

SDS-PAGE

70-80 kDa, under reducing conditions

Product Datasheets

AVI7194

Product Datasheet

COA

SDS

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.

AVI7194

Formulation	Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose.
Reconstitution	Reconstitute at 400 μg/mL in 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.

Scientific Data

Binding Activity

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When Biotinylated Recombinant Human IL-15R alpha Fc Chimera Avi-tag (Catalog # AVI7194) is immobilized at 0.5 µg/mL (100 µL/well), Recombinant Human IL-15 (247-ILB) binds with an ED₅₀ of 0.02-0.16 ng/mL.

SDS-PAGE

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2 μg/lane of Biotinylated Recombinant Human IL-15R alpha Fc Chimera Avi-tag (Catalog # AVI7194) was resolved with SDS-PAGE under reducing (R) and non-reducing (NR) conditions and visualized by Coomassie® Blue staining, showing bands at 70-80 kDa and 140-160 kDa, respectively.

Reconstitution Calculator

Background: IL-15R alpha

Interleukin 15 Receptor alpha (IL‑15 R alpha ), also known as CD215, is a widely expressed 60-kDa transmembrane glycoprotein that plays an important role in the homeostasis and activation of NK cells and CD8+ memory T cells and participates in the development and function of many other hematopoietic cell types and non‑immune cell types (1‑3). Mature human IL‑15 R alpha consists of a 175 amino acid (aa) extracellular domain (ECD) containing one N‑linked glycosylation site, a 23 aa transmembrane segment, and a 39 aa cytoplasmic tail (4). Within the ECD, human IL‑15 R alpha shares approximately 60% aa sequence identity with mouse and rat IL‑15 R alpha. Alternate splicing of human IL‑15 R alpha generates additional isoforms with variable length deletions in the ECD and/or substitutions in the cytoplasmic domain (4, 5). IL‑15 R alpha binds to Interleukin‑15 with high affinity (6). IL‑15 additionally interacts with lower affinity to a complex of IL‑2 R beta and the common gamma chain ( gamma c) which are also subunits of the IL‑2 receptor complex (7, 8). The use of shared receptor components contributes to the overlapping biological effects of IL‑15 and IL‑2. The dominant mechanism of IL‑15 action is known as transpresentation in which IL‑15/IL‑15 R alpha complexes are expressed on the surface of one cell and interact with complexes of IL‑2 R beta / gamma c on adjacent cells (9). This enables cells to respond to IL‑15 even if they do not express IL‑15 R alpha (10‑12). IL‑15/IL‑15 R alpha complexes can transmit reverse signaling that promotes cellular adhesion, tyrosine phosphorylation of intracellular proteins, and cytokine secretion by the IL‑15/IL‑15 R alpha expressing cells (13, 14). Shed soluble forms of IL‑15 R alpha retain the ability to bind tightly to IL‑15 and can inhibit IL‑15 bioactivity (6, 15, 16). Our Avi-tag Biotinylated Recombinant Human IL‑15 R alpha features biotinylation at a single site contained within the Avi-tag, a unique 15 amino acid peptide. Protein orientation will be uniform when bound to streptavidin-coated surface due to the precise control of biotinylation and the rest of the protein is unchanged so there is no interference in the protein's bioactivity.

References

Ma, A. et al. (2006) Annu. Rev. Immunol. 24:657.
Di Sabatino, A. et al. (2011) Cytokine Growth Factor Rev. 22:19.
Budagian, V. et al. (2006) Cytokine Growth Factor Rev. 17:259.
Anderson, D.M. et al. (1995) J. Biol. Chem. 270:29862.
Dubois, S. et al. (1999) J. Biol. Chem. 274:26978.
Giri, J.G. et al. (1995) EMBO 14:3654.
Grabstein, K. et al. (1994) Science 264:965.
Giri, J. et al. (1994) EMBO J. 13:2822.
Stonier, S.W. and K.S. Schluns (2010) Immunol. Lett. 127:85.
Duitman, E.H. et al. (2008) Mol. Cell. Biol. 28:4851.
Dubois, S. et al. (2002) Immunity 17:537.
Burkett, P.R. et al. (2004) J. Exp. Med. 200:825.
Budagian, V. et al. (2004) J. Biol. Chem. 279:42192.
Neely, G.G. et al. (2004) J. Immunol. 172:4225.
Budagian, V. et al. (2004) J. Biol. Chem. 279:40368.
Mortier, E. et al. (2004) J. Immunol. 173:1681.

Long Name

Interleukin 15 Receptor alpha

Entrez Gene IDs

3601 (Human); 16169 (Mouse); 102121571 (Cynomolgus Monkey)

Alternate Names

CD215 antigen; CD215; IL15 R alpha; IL-15 R alpha; IL-15 receptor subunit alpha; IL-15R alpha; IL15RA; IL-15Ra; IL-15R-alpha; interleukin 15 receptor, alpha; interleukin-15 receptor subunit alpha; MGC104179