CD40/TNFRSF5 Biologically Active Protein Summary
A biologically active protein corresponding to 173 residues of CD40.
Amino Acid Sequence: EPPTACREKQYLINSQCCSLCQPGQKLVSDCTEFTETECLPCGESEFLDTWNRETHCHQHKYCDPNLGLRVQQKGTSETDTICTCEEGWHCTSEACESCVLHRSCSPGFGVKQIATGVSDTICEPCPVGFFSNVSSAFEKCHPWTSCETKDLVVQQAGTNKTDVVCGRSSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK
A DNA sequence encoding the signal peptide and extracellular domain of human CD40 (aa
1-193) was fused to the Fc region of human IgG1 (aa 90-330). The chimeric protein was
expressed in modified human 293 cells.
|Details of Functionality
The ED50 of CD40 (173aa) Fc HCX Chimera is typically 1.0-1.5 ug/ml as measured by its ability to neutralize CD40 ligand mediated proliferation of T47-D cells.
Packaging, Storage & Formulations
Store at -80C. Avoid freeze-thaw cycles.
Reconstitute with 0.5 ml sterilized PBS containing 1% human serum albumin and 10% trehalose.
This protein is functionally active and can be used for Blocking and Neutralizing. It can also be used for Western Blot. CD40 (173aa) - Fc HCX Chimera separates into a number of glycoforms with an observed pI between 5.5 and 9.0 in 2D PAGE due to post-translational modifications, inparticular glycosylation. This compares with the unmodified CD40 (173aa) - Fc Chimera thathas a predicted pI of 6.0. CD40 (173aa) - Fc HCX Chimera contains N-linked oligosaccharides and may contain O-linked oligosaccharides. Under reducing conditions Symansis CD40 (173aa) - Fc HCX Chimera migrates as a broad band between 50 and 65 kDa on SDS-PAGE due to post-translational modifications, in particular glycosylation. This compares with unmodified CD40 (173aa) - Fc Chimera that has a predicted monomeric molecular mass of 46.5 kDa.
CD40 (TNFRSF5, CD40L Receptor) is a type I membrane protein and a member of the TNF receptor family. CD40 is expressed on B cells, monocytes, dendritic cells, endothelial cells, epithelial cells and B cell lymphomas and carcinomas. CD40 activates mulitple signalling pathways and can promote cell survival through up-regulation of the expression of genes encoding anti-apoptopic proteins. 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 CD40 (173aa) - Fc HCX Chimera consists of 5 to 30% carbohydrate by weight.
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 2 years from date of receipt.
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