human-GDF15-Qk017-protein-control-TGF-beta-activin-a-activity

Recombinant human GDF-15 protein (Qk017)

Name: Recombinant human GDF-15 protein (Qk017)
SKU: Qk017
Availability: InStock

Human GDF15 (growth differentiation factor 15) protein is a member of the TGFβ family and subject of intense interest as a marker of cellular stress and for its role in metabolism, cancer and pregnancy. Human GDF15 also is functional in mouse studies.

This 25 kDa disulfide-linked dimer is composed of the mature domain of human growth differentiation factor 15 protein. Our recombinant GDF15 protein is expressed in E. coli and exceptionally high purity extensively validated to ensure no trace contamination of related TGFβ family proteins from the mammalian culture systems, which historically led to controversy in the literature.

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1mg may be despatched as 2 x 500µg

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Description

Summary

High purity human GDF15 protein (Uniprot: Q99988)
25 kDa (dimer). Mature active protein is a disulfide-linked dimer.
Expressed in E. coli
Carrier protein-free
Manufactured in our Cambridge, UK laboratories
Lyophilized from acetonitrile, TFA
Resuspend in 10mM HCl at >100 µg/ml, prepare single use aliquots, add carrier protein if desired and store frozen at -20 oC or -80 oC

Handling and Storage FAQ

Featured applications

Biomarker for cellular stress
In vivo metabolic studies in mice (using human GDF15)

Alternative protein names

Growth/differentiation factor 15, Macrophage inhibitory cytokine 1 (MIC 1), NSAID activated gene 1 protein (NAG1)

Background references

1. Emmerson, P. J., Duffin, K. L., Chintharlapalli, S. & Wu, X. GDF15 and Growth Control. Front. Physiol. 9, 1712 (2018).
2. Olsen, O. E., Skjærvik, A., Størdal, B. F., Sundan, A. & Holien, T. TGF-β contamination of purified recombinant GDF15. PLoS One 12, e0187349 (2017).

Species reactivity

human

species similarity:
mouse – 67%
porcine – 67%
rat – 66%
bovine – 66%

Bioactivity

Human GDF15 Qk017 protein purity SDS-PAGE lot #011

GDF15 signals through GRAL and co-receptor RET leading to RET phosphorylation and signalling through the ERK and AKT pathway (reviewed in Emmerson et al., 2018). Commercial sources of GDF15, in particular those purified from mammalian expression systems, have been shown previously to be contaminated with trace amounts of TGFβ. These trace contaminants cause misleading experimental results due to the picomolar or even femtomolar EC50s (Olsen et al., 2017). Here we use a well-characterized SMAD2/3 activation assay to show that there is no contamination from other TGFβ family proteins.

Bioactivity is determined using a luciferase reporter assay in HEK293T cells. Cells are treated (in triplicate) with a serial dilution of GDF15 or Qk010 TGFβ1 for 6 hours. Firefly luciferase activity is measured and normalized to the control Renilla luciferase activity. EC50 = 0 pM (no contamination with TGFβ or related growth factors). Data from Qk017 lot #104282

Purity

GDF15 migrates as a single band at 24 kDa in non-reducing (NR) and 13 kDa as a single monomeric species upon reduction (R). No contaminating protein bands are visible.

Purified recombinant protein (7 µg) was resolved using 15% w/v SDS-PAGE in reduced (+β-mercaptothanol, R) and non-reduced conditions (NR) and stained with Coomassie Brilliant Blue R250. Data from Qk017 lot #010

Further quality assays

Mass spectrometry: single species with expected mass
Analytical reversed-phase: single sharp peak
Endotoxin: <0.005 EU/μg protein (below level of detection)
Recovery from stock vial: >95%

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download MSDS

We are a company founded and run by scientists to provide a service and support innovation in stem cell biology and regenerative medicine. All our products are exceptionally high purity, with complete characterisation and bioactivity analysis on every lot.

Protein background

Growth differentiation factor 15 (GDF-15) is a distant member of the TGFβ superfamily. Its expression is tightly regulated and circulating GDF15 protein in serum is associated with diseases such as cancer, cardiovascular disease, obesity and metabolic disease. GDF15 protein is being recognized as an important biomarker for cellular stress.

Unlike other members of the TGFβ superfamily that cause activation of the SMAD pathway, GDF15 protein signals through GRAL and co-receptor RET leading to RET phosphorylation and signalling through the ERK and AKT pathway (reviewed in Emmerson et al., 2018¹). Commercial sources of recombinant human GDF15 protein, in particular those purified from mammalian expression, are frequently contaminated with trace amounts of TGFβ and related proteins. These trace contaminants cause misleading experimental results due to the picomolar or even femtomolar EC50s of this family of cytokines². Please be cautious with your source of recombinant GDF15 protein, our scientists are happy to provide further information, please email support@qkine.com

We produce our proteins in E. coli with no animal products in our culture or purification processes to ensure there is no contamination from related proteins. In addition, we use a well-characterized SMAD2/3 activation assay to confirm there is no SMAD signalling.

Our products are for research use only and not for diagnostic or therapeutic use. Products are not for resale.

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