Recombinant Human IL-5R alpha/CD125 Protein

Product Discontinued

Recombinant Human IL-5R alpha/CD125 Protein Summary

• Description: A recombinant protein to IL5RA.
• Specificity: IL-5 R alpha-Fc HCX Chimera migrates as a broad band between 70 and 80 kDa in SDS-PAGE due to post-translation modifications, in particular glycosylation. This compares with the unmodified IL-5 R alpha-Fc Chimera that has a predicted molecular mass of 62.1 kDa. IL-5 R alpha-Fc HCX Chimera has N-linked and O-linked oligosaccharides.
• Preparation Method: A DNA sequence encoding the signal peptide and extracellular domain of human Interleukin 5 receptor alpha chain (aa 21-327) was fused to the Fc region of human IgG1 (aa 93-330). The chimeric protein was expressed in modified human 293 cells.
• Protein/Peptide Type: Recombinant Protein
• Gene: IL5RA

Applications/Dilutions

• Dilutions:
  • Block/Neutralize
  • Functional
  • Western Blot
• Application Notes: This protein is functionally active and can be used for Blocking and Neutralizing. It can also be used for Western Blot. IL-5 R alpha-Fc HCX Chimera separates into a number of isoforms with a pI between 5.8 and 7.0 in 2D PAGE due to post-translational modifications, in particular glycosylation. This compares with the unmodified IL-5 R alpha-Fc Chimera that has a predicted pI of 6.34.

Reactivity Notes

This is a Human protein

Packaging, Storage & Formulations

• Storage: Store at -80C. Avoid freeze-thaw cycles.
• Concentration: LYOPH
• Reconstitution Instructions: Reconstitute with 0.5 ml sterilized PBS. After reconstitution the buffer solution will contain 1% human serum albumin and 10% trehalose already present in the vial.

Notes

Purity is greater than 95%, as determined by SDS-PAGE and visualized by silver stain.

Alternate Names for Recombinant Human IL-5R alpha/CD125 Protein

• CD125 antigen
• CD125
• CDw125CD125
• HSIL5R3
• IL-5 R alpha
• IL-5 receptor subunit alpha
• IL5R alpha
• IL-5R subunit alpha
• IL5R
• IL5RA
• IL-5Ra
• IL-5R-alpha
• interleukin 5 receptor type 3
• interleukin 5 receptor, alpha
• interleukin-5 receptor alpha chain
• interleukin-5 receptor alpha subunit
• interleukin-5 receptor subunit alpha
• MGC26560

Background

Interleukin 5 (IL-5) plays an important role in B cell development by inducing the terminal differentiation of late-developing B cells. Additionally, IL-5 regulates the differentiation, survival and degranulation of eosinophils. IL-5 is expressed predominately by T-lymphocytes and mast cells, and to a lesser extent by eosinophils, natural killer cells and endothelial cells. IL-5 specifically binds to the IL-5 receptor-alpha (IL-5 R alpha; IL-5Ra) subunit expressed predominately on CD5+ B cells, eosinophils, mast cells, CD34+ stem cells, and basophils. Binding allows association with the common beta subunit (Beta c), shared by the IL-3 and GM-CSF receptors. In addition to the membrane bound form of IL-5Ra, soluble forms of IL-5Ra have been identified and are inhibitory to IL-5 activity. There is also evidence to suggest that the levels of membrane bound and soluble IL-5 receptors are altered in disease states such as asthma, where there is an increase in the membrane bound form and a concomitant decrease in the soluble receptor form. IL-5 R alpha is produced as an ECD-Fc fusion protein with the aim of enhancing its activity. ECD-Fc fusion proteins have an advantage over soluble receptors because many receptors are only functional in dimeric form. Fusion to the Fc domain of IgG1 induces dimerization due to the ability of the Fc domain to form disulfide bonds. The resulting dimeric receptor (EDC-Fc) mimics the activated form of the receptor and possesses enhanced affinity for its cognate ligand relative to its monomeric form. This is a HCX protein. HCX Expression System Details HCX proteins mimic the proteins in the human body because they are expressed from human, rather than animal, insect or bacterial cells. This process gives them human post-translational modifications. Recombinant DNA techniques allow a human protein with the correct amino acid sequence to be expressed in a non-human cell line. However, non-human cells lack the appropriate cellular machinery, such as specific glycosyltransferases, necessary to produce the correct human post-translational modifications of a protein. An extreme example is seen in E. coli cells, which produce recombinant proteins with no glycosylation, as the above figure illustrates. Rodent and yeast cells are able to glycosylate proteins, but they are still different from glycosylation in human cells. Expression System Resultant Proteins Human (e.g. K562, HEK293) Correct amino acid sequence Human post-translational modifications Rodent (e.g. CHO, NSO) Correct amino acid sequence Some natural glycosylation - not human-like Yeast (e.g. Pichia) Correct amino acid sequence Some natural glycosylation - not human-like E.Coli Correct amino acid sequence No PTMs Although there have been significant attempts to make non-human cell derived cytokines more human-like, there is a growing awareness that in many instances, particularly in therapeutics, cytokines should mimic those found in the body as closely as possible. Purified IL-5 R alpha-Fc HCX Chimera consists of 10-25% carbohydrate by weight.

Limitations

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Peptides and proteins are guaranteed for 3 months from date of receipt.

Bioinformatics

• Gene Symbol: IL5RA