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GM-CSF Reporter HEK 293 Cells

HEK-Blue™ GM-CSF Cells Unit size Cat. code Docs Qty Price
Human GM-CSF Reporter Cells
3-7 x 10e6 cells
hkb-hgmcsfr
+-
$1,304.00

You may also need : HEK-Blue™ Selection | View more associated products

HEK-Blue™ GM-CSF Cells signaling pathway
HEK-Blue™ GM-CSF Cells signaling pathway

Granulocyte-Macrophage Colony-Stimulating Factor Reporter Cells

HEK-Blue™ GM-CSF cells were engineered from the human embryonic kidney HEK 293 cell line to detect bioactive granulocyte-macrophage colony-stimulating factor (GM-CSF, aka CSF2) by monitoring the activation of the JAK2/STAT5 pathway.  Although originally identified as a hematopoietic growth factor, this cytokine is now regarded as a pleiotropic regulator of inflammation in the responses to pathogens, autoimmune diseases, and cancer [1, 2].

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Cell line description:

HEK-Blue™ GM-CSF cells were generated by stable overexpression of the genes encoding for the human GMRα (CD116), GMRβ (CD131), and STAT5, as well as a STAT5-inducible secreted embryonic alkaline phosphatase (SEAP) reporter. STAT5-dependent SEAP activity is readily assessable in the supernatant using QUANTI-Blue™ Solution, a detection reagent.

Features of HEK-Blue™ GM-CSF cells:

  • Fully functional GM-CSF signaling pathway
  • Readily assessable STAT5-inducible SEAP reporter activity
  • The stability for 20 passages has been verified
  • Functionally tested and guaranteed mycoplasma-free

Applications of HEK-Blue™ GM-CSF cells:

  • Detection of human GM-CSF (5 pg/ml - 100 ng/ml)
  • No detection of murine GM-CSF
  • Screening of anti-GM-CSF and anti-GMRα antibodies

 

References:

1. Dougan M. et al., 2019. GM-CSF, IL-3, and IL-5 family of cytokines: regulators of inflammation. Immunity. 50(4):796-811.
2. Zhan Y. et al., 2019. The Pleiotropic Effects of the GM-CSF Rheostat on Myeloid Cell Differentiation and Function: More Than a Numbers Game. Front Immunol. 102679.
3. Hamilton J.A., 2020. GM-CSF in inflammation. J. Exp . Med. 217(1):e20190945.

Figures

Validation of GMRα expression
Validation of GMRα expression

Validation of the expression of human GMRα by HEK-Blue™ GM-CSF cells.
5 x 105 cells were incubated with either an APC-conjugated isotype control (blue) or an APC-conjugated Anti-GMRα mAb (red) for 1 hour. The binding affinity was then measured using flow cytometry.

Cellular response to GM-CSF
Cellular response to GM-CSF

Dose-response of HEK-Blue™ GM-CSF cells to recombinant GM-CSF.
Cells were stimulated with increasing concentrations of recombinant human GM-CSF (hGM-CSF) and murine GM-CSF (mGM-CSF). After overnight incubation, the STAT5 response was determined using QUANTI‑Blue™ Solution, a SEAP detection reagent. The optical density (OD) at 630 nm is shown as mean ± SEM.

Cellular response to GM-CSF
Cellular response to GM-CSF

Response of HEK-Blue™ GM-CSF cells to a panel of cytokines.
Cells were stimulated with various human and murine recombinant cytokines: 0.1 ng/ml of hGM-CSF, 10 ng/ml mGM-CSF, hIL‑6, hIL-27, or 1000 U/ml hIFN-α. After overnight incubation, SEAP activity was assessed using QUANTI‑Blue™ Solution. The optical density (OD) at 630 nm is shown as mean ± SEM.

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Specifications

Antibiotic resistance: Blasticidinhygromycin, and  Zeocin™

Growth medium: DMEM, 4.5 g/l glucose, 2 mM L-glutamine, 10% (v/v) heat-inactivated fetal bovine serum (FBS;  30 min at 56°C), 100 U/ml penicillin, 100 μg/ml streptomycin, 100 μg/ml Normocin™

Specificity: human GM-CSF

Detection range:  5 pg/ml - 100 ng/ml for human GM-CSF

Quality Control:

  • STAT5-dependent SEAP reporter activity in response to human GM-CSF has been validated.
  • The cell surface expression of human GMRα  (CD116) in this cell line has been validated using fluorescence-activated cell sorting (FACS).
  • The stability for 20 passages, following thawing, has been verified. 
  • These cells are guaranteed mycoplasma-free. 

This product is covered by a Limited Use License (See Terms and Conditions).

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Contents

  • 3-7 x 106 HEK-Blue™ GM-CSF cells in a cryovial or shipping flask
  • 1 ml Normocin™ (50 mg/ml)
  • 2 x 1 ml of HEK-Blue Selection (250X)
  • 1 ml of QB reagent and 1 ml of QB buffer (sufficient to prepare 100 ml of QUANTI-Blue™ Solution, a SEAP detection reagent).

Shipped on dry ice (Europe, USA & Canada)

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Details

The granulocyte-macrophage colony-stimulating factor (GM-CSF, aka CSF2) belongs to the β-common chain cytokine family [1]. Although originally identified as a hematopoietic growth factor, this cytokine is now regarded as a pleiotropic regulator of inflammation in the responses to pathogens, autoimmune diseases, and cancer [1, 2]. GM-CSF signalization requires a multimeric structure comprising four α chains (GMRα, aka CD116), four β chains (GMRβ aka CD131), and four cytokines. This 12-protein complex allows the juxtaposition of the intracellular Janus kinase 2 (JAK2) and activation of the signal transducer and activator of transcription 5 (STAT5) [1]. Other signaling pathways include ERK, NF-κB, and AKT pathways [2, 3]. 

The understanding of cellular and molecular mechanisms whereby GM-CSF exerts its varied functions is key for the development of therapeutic strategies (e.g. cancer vaccines, blocking antibodies) [1]. 

 

1. Dougan M. et al., 2019. GM-CSF, IL-3, and IL-5 family of cytokines: regulators of inflammation. Immunity. 50(4):796-811.
2. Zhan Y. et al., 2019. The Pleiotropic Effects of the GM-CSF Rheostat on Myeloid Cell Differentiation and Function: More Than a Numbers Game. Front Immunol. 102679.
3. Hamilton J.A., 2020. GM-CSF in inflammation. J. Exp . Med. 217(1):e20190945.

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FAQ

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