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Recombinant Human CD52 Fc Chimera Avi-tag Protein, CF

Catalog #: AVI9116 Datasheet / COA / SDS
Biotinylated
Catalog # Availability Size / Price Qty
AVI9116-050
Recombinant Human CD52 Fc Chimera Avi-tag Protein Bioactivity.
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Summary Product Datasheets Carrier Free Data Images Reconstitution Calculator Background Related Research Areas

Recombinant Human CD52 Fc Chimera Avi-tag Protein, CF Summary

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
Measured by its binding ability in a functional ELISA. When Recombinant Human SIGLEC-10 Fc Chimera Protein  (Catalog # 2130-SL) is immobilized at 5.0 µg/mL (100 µL/well), the concentration of Biotinylated Recombinant Human CD52 Fc Chimera Avi-tag (Catalog # AVI9116) that produces 50% of the optimal binding response is 0.500-5.00 µg/mL.
Source
Chinese Hamster Ovary cell line, CHO-derived human CD52 protein
Human CD52
(Gly25-Ser36)
Accession # P31358.1		IEGRMDHuman IgG1
(Pro100-Lys330)		Avi-tag
N-terminusC-terminus
Accession #
P31358.1
N-terminal Sequence
Analysis
Ser24 & Gly25
Structure / Form
Disulfide-linked homodimer
Biotinylated via Avi-tag
Predicted Molecular Mass
30 kDa
SDS-PAGE
38-44 kDa, under reducing conditions.

Product Datasheets

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AVI9116

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.

AVI9116

Formulation Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose.
Reconstitution Reconstitute at 250 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

Bioactivity View Larger

When Recombinant Human SIGLEC-10 Fc Chimera Protein (2130-SL) is immobilized at 5.0 µg/mL (100 µL/well), the concentration of Biotinylated Recombinant Human CD52 Fc Chimera Avi-tag (Catalog # AVI9116) that produces 50% of the optimal binding response is 0.500-5.00 µg/mL.

SDS-PAGE View Larger

2 μg/lane of Recombinant Human CD52 Fc Chimera Avi-tag Protein (Catalog # AVI9116) was resolved with SDS-PAGE under reducing (R) and non-reducing (NR) conditions and visualized by Coomassie® Blue staining, showing bands at 38-44 kDa and 80-90 kDa, respectively.

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:

CD52, also known as CAMPATH-1 antigen, HE5, and gp20, is a cell surface glycoprotein that can be targeted to induce immune suppression by complement-mediated cell lysis (1, 2). Mature human CD52 is a 12 amino acid peptide that is tethered to the cell surface with a GPI linkage (3). It carries a large tissue-specific carbohydrate structure which increases the molecular weight of CD52 to 15-20 kDa (3-5). CD52 is expressed on lymphocytes, monocytes, monocycte-derived dendritic cells, eosinophils, and neutrophils (6-8), as well as on mature spermatozoa and epithelial cells lining the male genital tract (5, 9, 10). It protects sperm against anti-sperm antibodies by binding to C1q and inhibiting complement activation (11). CD52 ligation on CD4+ T cells induces a suppressive population of cells that release soluble CD52 which then binds to Siglec-10 (12-14). This interaction inhibits the proliferation of activated T cells and the cytotoxic function of autoimmune CD8+ T cells in type 1 diabetes (13). Our Avi-tag Biotinylated human CD52 Fc chimera 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
  1. Morris, P.J. and N.K. Russell (2006) Transplantation 81:1361.
  2. Redpath, S. et al. (1998) Immunology 93:595.
  3. Xia, M.Q. et al. (1991) Eur. J. Immunol. 21:1677.
  4. Treumann, A. et al. (1995) J. Biol. Chem. 270:6088.
  5. Schroter, S. et al. (1999) J. Biol. Chem. 274:29862.
  6. Ratzinger, G. et al. (2003) Blood 101:1422.
  7. Elsner, J. et al. (1996) Blood 88:4684.
  8. Ambrose, L.R. et al. (2009) Blood 114:3052.
  9. Focarelli, R et al. (1998) Mol. Hum. Reprod. 4:119.
  10. Hale, G. et al. (1993) J. Reprod. Immunol. 23:189.
  11. Hardiyanto, L. et al. (2012) J. Reprod. Immunol. 94:142.
  12. Watanabe, T. et al. (2006) Clin. Immunol. 120:247.
  13. Bandala-Sanchez, E. et al. (2013) Nat. Immunol. 14:741.
  14. Toh, B.-H. et al. (2013) Cell. Mol. Immunol. 10:379.

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