GM- CSF Human

GM- CSF Human

CSF

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  • Name :

    GM- CSF Human

    Description:

    Granulocyte Macrophage-Colony Stimulating Factor Human Recombinant

    CSF-2, MGI-1GM, GM-CSF, Pluripoietin-alpha, MGC131935, MGC138897.

    Product # :

    CYT-221

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  • description
  • source
  • formulation
  • purity
  • biological activity
  • More Info

Description

Granulocyte Macrophage Colony Stimulating Factor Human Recombinant produced in E.Coli is a single, non-glycosylated, polypeptide chain containing 127 amino acids and having a molecular mass of 14477 Dalton. GM-CSF is purified by proprietary chromatographic techniques.

Source

Escherichia Coli.

Formulation

GM-CSF was lyophilized after extensive dialysis against 2mM sodium phosphate buffer pH= 7.4±0.1.

Purity

Greater than 98.0% as determined by:
1. Analysis by RP-HPLC.
2. Analysis by SDS-PAGE.

Biological Activity

The ED50 as determined by the dose-dependant stimulation of the proliferation of human TF-1 cells (human erythroleukemic indicator cell line) is < 0.1 ng/ml, corresponding to a Specific Activity of 11,100,000 IU/mg.

More Info

  • Introduction

    Granulocyte Macrophage Colony Stimulating Factor (GM-CSF) was first characterized as a growth factor that supports the in-vitro colony formation of granulocytes-macrophages progenitor cells. It is a pleiotropic cytokine and a member of a family of endogenous cytokines of the hematopoietic system. GM-CSF is produced as a response to immune or inflammatory stimuli by activated cells of the hematopoietic system such as T cells, B cells, macrophages, mast cells and also fibroblasts and alveolar epithelial cells. It plays an important role in regulating the proliferation, differentiation, survival and activation of hematopoietic cells such as granulocytes and monocytes ,neutrophiles, basophiles and eosonophoiles, erythroid cells, megakaryocytes and T cells.
    Human and mouse GM-CSF have about 56% homology and are species specific. Human GM-CSF is not active on mouse cells and vice versa. It is active on canine and feline cells.
    GMCSF is 144 amino acids, 22kDa glycoprotein. It is composed of four bundles alpha helices. Its receptor is heterodimers with a ligand-specific alpha subunit and a betac subunit that is shared with the interleukin IL-3 and IL-5 receptors. This unusual form of receptor assembly likely applies also to IL-3 and IL-5 receptors. Cross-linking the two receptor subunits is required for receptor activation and signaling .
    GMCSF has been shown to be involved in maturation, mobilization and antigen presentation of myeloid dentritic cells (DCs) in-vivo or ex-vivo. This function promotes Th1 immune responses, cytotoxcity, anti-angiogenesis as well as allergic inflammation, and the development of autoimmunity. Therefore GMCSF can be used in immunotherapy for the treatment of immune suppressed and immune-compromised patients as well as in veterinary medicine for the same purpose. GM-CSF is also important in regulation of embryo development and pregnancy and specifically in embryo implantation and subsequent development .

  • Synonyms

    CSF-2, MGI-1GM, GM-CSF, Pluripoietin-alpha, MGC131935, MGC138897.

  • Physical Appearance

    Sterile Filtered White lyophilized (freeze-dried) powder.

  • Stability

    Lyophilized Granulocyte Macrophage Colony Stimulating Factor although stable at room temperature for 3 weeks, should be stored desiccated below -18°C. Upon reconstitution GMCSF should be stored at 4°C between 2-7 days and for future use below -18°C.For long term storage it is recommended to add a carrier protein (0.1% HSA or BSA).Please prevent freeze-thaw cycles.

  • Solubility

    It is recommended to reconstitute the lyophilized Granulocyte Macrophage Colony Stimulating Factor in sterile 18MΩ-cm H2O not less than 100µg/ml, which can then be further diluted to other aqueous solutions.

  • Amino Acid Sequence

    The sequence of the first five N-terminal amino acids was determined and was found to be Ala-Pro-Ala-Arg-Ser.
    N-terminal methionine has been completely removed enzymatically.

  • Background

    What is the molecular weight/Mw of GM CSF HUMAN Protein?
    GM CSF HUMAN Protein has a total Mw of 14.47kDa.

    What is the source or expression system of GM CSF HUMAN Protein?
    Escherichia Coli.

    What is the Purity of GM CSF HUMAN Protein?
    GM CSF HUMAN Protein is >98% pure as determined by SDS-PAGE.

    What is the Biological Activity of GM CSF HUMAN Protein?
    The ED50 as determined by the dose-dependant stimulation of the proliferation of human TF-1 cells (human erythroleukemic indicator cell line) is < 0.1 ng/ml, corresponding to a Specific Activity of 11,100,000 IU/mg.

    What is the amino acid sequence of GM CSF HUMAN Protein?
    The sequence of the first five N-terminal amino acids was determined and was found to be Ala-Pro-Ala-Arg-Ser.
    N-terminal methionine has been completely removed enzymatically.

    What applications can GM CSF HUMAN Protein be used in?
    GM CSF HUMAN Protein can probably be used in western blot, ELISA and Lateral Flow.

    What is the endotoxin level for GM CSF HUMAN Protein?
    The endotoxin level is minimal, GM CSF HUMAN Protein was purified using conventional chromatography techniques.


  • Protein content

    GM-CSF quantitation was carried out by two independent methods:1. UV spectroscopy at 280 nm using the absorbency value of 0.963 as the extinction coefficient for a 0.1% (1mg/ml) solution. This value is calculated by the PC GEN computer analysis program of protein sequences (IntelliGenetics). 2. Analysis by RP-HPLC, using a standard solution of GM-CSF as a Reference Standard.

ProSpec's products are furnished for LABORATORY RESEARCH USE ONLY. The product may not be used as drugs, agricultural or pesticidal products, food additives or household chemicals.

Gm Csf Human

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Frequently asked questions

1. Which company offers Recombinant Growth Factors at competitive prices?

ProSpec offers vast array of Growth Factors such as EGF| IGF1 | Leptin| VEGF and much more at affordable descent prices to meet academic budgets
ProSpec provides High Purity Growth Factors for 30 years, purity exhibits 95% and more as visualized by SDS-PAGE gel.
Growth Factors are in stock for immediate delivery via FedEx which enables scientists and researchers to perform immediate experiments.
Growth Factors are shipped globally using ProSpec’s custom tailored Styrofoam insulated box and premium ice packs that ensure maximum protein stability during transit to your laboratory.

2. Who can produce custom Growth Factor for me?

ProSpec offers custom production of recombinant Growth Factors, managing the entire process from gene design and cloning to protein expression and fermentation. Harvest and cell breakage. Purification using standard chromatography, quality control testing, and formulation. Freeze-drying (lyophilization) is available as an option. For more information contact us at info@prospecbio.com

3. How long does it take to produce a custom Growth Factor

The production time line for a custom recombinant growth factor can take about 5-7 weeks.

4. Where can i find biologically active Growth Factors?

ProSpec offers biologically functional Growth Factors such as Prolactin | Erythropoietin | PDGF-BB which are rigorously tested in-vitro to provide researchers world wide BioActive Growth Factors.

5. What is the activity validation method for Growth Factors?

ProSpec uses cell-based bioassays that measures the biological activity, for example:
Activin-A is validated by Activin-responsive luciferase assay, SMAD2/3 phosphorylation.
CSF, G-CSF, GM-CSF, M-CSF are validated by Cytokine-dependent cell proliferation assay.
CTGF is validated by Fibroblast proliferation or migration assay.
EGF is validated by NIH3T3 or Balb/3T3 proliferation assay.
Epigen (EPGN) is validated by EGFR activation assay, cell proliferation.
Erythropoietin (EPO) is validated by TF-1 or UT-7 erythroid cell proliferation assay.
Fibroblast Growth Factors is validated by NIH3T3 proliferation assay.
IGF-1 and IGF-2 are validated by Cell proliferation assay (MCF-7, Saos-2, BALB/3T3).

6. Is the Growth Factor full length or partial?

The Growth Factor proteins are both full length and partial while majority of them are active mature proteins rather than a full-length precursor. ProSpec active mature proteins exhibits higher expression yields, better stability, Easier purification and equivalent biological activity as full length precursor.

7. Is the Growth Factor monomeric or dimeric form?

This depends on the Growth Factor. ProSpec Growth Factors are supplied in the native biologically active oligomeric state, which can be monomeric, dimeric, trimeric, or even multimeric since because receptor binding and biological activity often depend on correct oligomerization.
The EGF, Epigen, Fibroblast Growth Factors (FGF1, FGF2, FGF7/KGF)
HDGF, IGFBPs (IGFBP-1 to IGFBP-7), IGF2 are usually provided as monomer proteins though Activin-A and myostatin are supplied as homodimers.

8. What is the Growth Factor purity and how was it determined?

The Growth Factors purity ranges from high purity of ≥95% to premium grade ≥97%.
ProSpec uses SDS-PAGE Coomassie Blue Staining as the primary purity method which Visualizes protein bands. Some SDS-PAGE gels are stained with Silver Stain. Other analytical methods include RP-HPLC [ High-resolution purity determination ] , SEC-HPLC for the detections of aggregates and fragments and Western Blot to confirm Growth Factors identity.

9. Is the Growth Factor sequence verified?

The amino acid sequence verification is performed at DNA & protein level.
Sanger DNA Sequencing verifies the expression plasmid having correct coding.
LC-MS is used to verify the molecular weight of the final purified Growth Factor
Prospec also uses Edman N-terminal Sequencing to determine the amino-terminal sequence.

10. What is the endotoxin level in Growth Factors and how do you test LPS ?

ProSpec Growth Factors are tested for endotoxin also named LPS [ lipopolysaccharide ] which is found in the outer membrane of Gram-negative bacteria which can affect cytokine secretion, cell proliferation and stem cell.
ProSpec measures endotoxin using the Limulus Amebocyte Lysate [ LAL ] assay such as Gel-Clot.
ProSpec standard Endotoxin level for cytokines is usually <1 EU/ug.
What Does "EU" Mean? EU = Endotoxin Units

11. Is there host cell protein contamination?

There might be a minor existence of host-cell proteins post chromatography purification and quality-control procedures that greatly minimize these impurities. ProSpec tests host-cell protein levels by SDS-PAGE silver staining, or mass spectrometry.

12. Is the protein tested in vitro, in-vivo cell culture, stem cells, T cells, ELISA, western blot?

PoSpec Growth Factors proteins are usually tested for biological activity and identity, while some proteins are not though supported by literature and customer experience. Interleukins are tested in T-cell, NK-cell, PBMC, stem cell culture. Interferons are tested in Antiviral assays, immune-cell activation, TNFA proteins are tested in Apoptosis, inflammation and immune studies. ProSpec does not conduct in-vivo or animal injections experiments in house or our source.