Molecular weight: rHuman GM-CSF is a glycosilated polypeptide chain containing 127 amino acids (18-144 aa CSF2_HUMAN P04141 ), fused to 10 His tag at N-terminal. rHuman GM-CSF migrate as a broad band between 15 and 25 kDa due to post-translation modification, in particular glycosilation.
Description: GMCSF is a cytokine that stimulate the growth and differentiation of hematopoietic precursor cells from various lineages, including granulocytes, macrophages, eosinophils and erythrocytes. Is involved in differentiation of dendritic cells and is a key factor in differentiation pathways leading form stem cells. GMCSF is produced by several cell types as monocytes, fibroblast, endothelial cells and T- Lymphocytes in response to a number of inflammatory mediators present in the hemopoietic environment and peripheral site of inflammation. Human GMCF is an important therapeutic cytokine used in the treatment of myeloid leukemia, neutropenia and aplastic anemia and it could become interesting in the treatment following bone marrow transplantation. It performs biological activity by binding to a receptor specific receptor complex which is composed of a cytokine-specific alpha chain and B chain shared with the receptors for interleukin-3 and interleukin-5. GMCSR has been identified to mediate the activation of Jak-Stat and MAPK pathways.
Fig 1. SDS-PAGE analysis of recombinant GM-CSF. Samples were loaded in 15% SDS-polyacrylamide gel and stained with Coomassie blue. Lane MWM: Molecular weight marker (kDa); lane 1 contains 500 ng of rhuman GM-CSF . All bands shown in lane 1 have been identify by MALDI-TOFF as recombinant GM-CSF.
Fig 2. Analysis of recombinant GM-CSF with specific anti-human GM-CSF by Western Blot; lane MWM: Molecular weight marker (kDa); lane 1 contains 500 ng of rhuman GM-CSF . All bands shown in lane 1 have been identify by MALDI-TOFF as recombinant GM-CSF.
Biological Activity:
1. The activity of recombinant human GM-CSF is determined by the dose-dependent induction of human TF-1 proliferation cell. ED50 is ≤ 0.05 ng/mL.
2. Effect of rH GM-CSF on kerotinocytes migration (HaCaT cells). 24 hours effect on skin cell migration, scale bar 100 px, A: Untreated cells; B: Positive control GF cocktail 10% ; C: 0.5 ng/ml rH GM-CSF; D: 1ng/ml rH GM-CSF.
3. Collagen I expression profile performed on keratinocytes (HaCaT cells). Expression of messenger RNA codifying for Collagen I in HaCaT cell treated with rh GM-CSF after 24 and 48 hours.
4. Effect of rh GM-CSF on Human fibroblast cell proliferation. Cell viability was assessed by MTT assay and showed very significant (*) (p<0.0001) stimulation of fibroblasts proliferation at 0,5 ng/ml and 1ng/ml. 
Human recombinant protein expressed in Nicotiana benthamiana.
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