Recombinant Human G-CSF Protein

Product Discontinued

Recombinant Human G-CSF Protein Summary

• Description: A biologically active protein to CSF3.
• Specificity: G-CSF migrates as a band between 15 and 20 kDa in SDS-PAGE. This compares with the predicted molecular mass of 18.7 kDa.
• Preparation Method: A DNA sequence encoding the human G-CSF protein sequence (containing the signal peptide sequence, and the mature G-CSF sequence) was expressed in modified human 293 cells.
• Details of Functionality: The ED50 of G-CSF is typically 0.01 - 0.03 ng/ml as measured in a cell proliferation assay using a murine myeloblastic M-NFS-60 cell line.
• Protein/Peptide Type: Biologically Active Protein
• Gene: CSF3

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

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.

Alternate Names for Recombinant Human G-CSF Protein

• C17orf33
• chromosome 17 open reading frame 33
• colony stimulating factor 3 (granulocyte)
• CSF3
• CSF3OS
• Filgrastim
• GCSF
• G-CSF
• GCSFlenograstim
• granulocyte colony-stimulating factor
• Lenograstim
• MGC45931
• Pluripoietin

Background

Granulocyte colony stimulating factor (G-CSF) is the primary extracellular regulator of granulopoiesis and regulates the production of neutrophils by stimulating proliferation and survival of specific bone marrow precursor cells and their differentiation into granulocytes. Neutrophils play a critical role in the defence against bacterial and fungal infections. G-CSF is produced by monocytes, macrophages, neutrophils, fibroblasts and endothelial cells and is capable of increasing the absolute number of circulating neutrophils and enhancing their antimicrobial function. Unlike GMCSF, the activity of G-CSF is not species specific. Additionally, G-CSF production is inducible by cytokines including TNF-alpha, IL-1, GM-CSF, IL-17 and IL-4. Granulocyte colony stimulating factor (G-CSF) is a glycoprotein consisting of 207 amino acids. Differential splicing of G-CSF mRNA can result in two protein variants either 177- or 180-amino acids in length. G-CSF contains 5 cysteine residues, four of which form intermolecular disulfide bonds. There is a potential O-glycosylation site at Thr-166, which inhibits aggregation and stabilises the molecule. 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. G-CSFhcx separates into a number of isoforms with a pI between 5.4 and 6.0 in 2D PAGE Due to post-translational modifications, in particular glycosylation,. This compares with the unmodified G-CSF that has a predicted pI of 5.65.

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 2 years from date of receipt.

Bioinformatics

• Gene Symbol: CSF3