Human LC3B Reporter THP-1 Cells

RFP::GFP::LC3-OE reporter monocytes

ABOUT

Monitoring autophagy activity 

THP1-Difluo™ hLC3 reporter cells are designed to monitor the autophagic flux by means of a fluorescent-labeled LC3B protein.

These cells, derived from the THP-1 human monocytic cell line, express the RFP::GFP::LC3 fusion protein in which the human LC3B (microtubule-associated protein 1 light chain 3 beta) is fused to two fluorescent reporter proteins: RFP (acid-stable) and GFP (acid-sensitive). The LC3B protein is a good marker to assess the autophagic flux since it is recruited from the cytosol to be associated to all the autophagic structures: the nascent autophagosome (called the isolation membrane), the complete autophagosome and the autolysosome. Upon autophagy induction, diffuse cytoplasmic fluorescence shifts to autophagosomes and autolysosomes as fluorescent puncta.

Monitoring of autophagy activity relies on microscopy visualization of fluorescent puncta, progressive degradation of the GFP, and concurrent increase of RFP signal in autolysosomes. The number of fluorescent punctate structures per cell, the percentages of RFP-GFP positive and RFP positive cells can be used to assess autophagic flux as described previously [1,2].

 

More details Read our review on Autophagy and innate immunity.

 

References

1. Loos B. et al. 2014. Defining and measuring autophagosome flux- concept and reality. Autophagy. 2014;10(11):2087-96. 
2. Kimura S. et al., 2007. Dissection of the autophagosome maturation process by a novel reporter protein, tandem fluorescent- tagged LC3. Autophagy. 3(5):452-60.

Disclaimer:  These cells are for internal research use only and are covered by a Limited Use License (See Terms and Conditions). Additional rights may be available.

SPECIFICATIONS

Specifications

Species
Human
Target

Autophagy

Tested applications

Monitoring autophagy activity

Cell type
Monocytic
Growth properties
Suspension
Tissue origin
Human monocytes
Reporter gene
GFP
RFP
Detection method
Fluorescence
Antibiotic resistance
Zeocin®
Growth medium

Complete RPMI 1640 (see TDS)

Mycoplasma-free

Verified using Plasmotest™

Quality control

Each lot is functionally tested and validated.

CONTENTS

Contents

  • Product: 
    THP1-Difluo™ hLC3 Cells
  • Cat code: 
    thpdf-hlc3
  • Quantity: 
    3-7 x 10^6 cells
Includes:
  • 1 ml of Zeocin® (100 mg/ml)
  • 1 ml of Normocin™ (50 mg/ml). Normocin™ is a formulation of three antibiotics active against mycoplasmas, bacteria, and fungi.

Shipping & Storage

  • Shipping method:  Dry ice
  • Storage:

    • Liquid nitrogen vapor
    Stability: 20 passages

    Caution:

    • Upon receipt, store immediately in liquid nitrogen vapor. Do not store cell vials at -80°C.

DOCUMENTS

Documents

THP1-Difluo™ hLC3 Cells

Technical Data Sheet

Safety Data Sheet

Validation Data Sheet

Certificate of analysis

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CUSTOMER SERVICE & TECHNICAL SUPPORT

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