Recombinant Human GM-CSF Protein

Product Discontinued

Recombinant Human GM-CSF Protein Summary

• Description: A biologically active protein to CSF2.
• Specificity: GM-CSFB contains N-linked and possibly O-linked oligosaccharides. GM-CSFB migrates as a broad band between 15 and 30 kDa in SDS-PAGE due to post-translation modifications, in particular glycosylation. This compares with unmodified GM-CSF that has a predicted molecular mass of 14.5 kDa.
• Preparation Method: A DNA sequence encoding the human GM-CSF protein sequence (containing the signal peptide sequence, and the mature GM-CSF sequence) was expressed in modified human 293 cells.
• Details of Functionality: The ED50 of GM-CSFB is typically 0.02 - 0.2 ng/ml as measured in a cell proliferation assay using the human growth factor-dependent TF-1 cell line.
• Protein/Peptide Type: Biologically Active Protein
• Gene: CSF2

Applications/Dilutions

• Dilutions:
  • Block/Neutralize
  • Functional
  • Western Blot
• Application Notes: This protein is functionally active and can be used for Blocking and Neutralizing. GM-CSFB separates into a number of isoforms with a pI between 3.5 and 5.7 in 2D PAGE due to post-translational modifications, in particular glycosylation. This compares with the unmodified GM-CSF that has a predicted pI of 5.21.
• Theoretical MW: 16 kDa.
  Disclaimer note: The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors.

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 GM-CSF Protein

• colony stimulating factor 2 (granulocyte-macrophage)
• Colony-stimulating factor
• CSF
• CSF2
• CSF-2
• GMCSF
• GM-CSF
• GMCSFgranulocyte-macrophage colony-stimulating factor
• granulocyte-macrophage colony stimulating factor
• MGC131935
• MGC138897
• Molgramostim
• molgramostin
• Sargramostim

Background

Granulocyte-macrophage colony stimulating factor (GM-CSF) regulates hematopoiesis, cell migration and immunity. GM-CSF is crucial for the growth and development of granulocyte and macrophage progenitor cells particularly during host defence and inflammatory reactions. GM-CSF is glycosylated at two N-linked and several O-linked sites. Glycosylation of GMCSF is important as it can affect pharmacokinetics, receptor affinity, biological activity, in vivo clearance rates and immunoreactivity. Patients receiving human recombinant GM-CSF commonly develop antibodies that recognize recombinant human GM-CSF produced in yeast (sargramostim) and E. coli (molgramostin), which exhibit glycosylation patterns that are distinct to the native human glycosylation. 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 GM-CSFB consists of 5-50% 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 1 year from date of receipt.

Bioinformatics

• Gene Symbol: CSF2