Mouse ASC-expressing RAW 264.7 Cells

ASC-OE mouse macrophages

ABOUT

ASC expressing RAW 264.7 cells

ASC (apoptosis-associated speck-like protein containing a CARD domain, also known as PYCARD) is a protein adaptor important in canonical inflammasome responses [1].

More details

 

To foster studies on the ASC adaptor, InvivoGen has developed RAW-ASC cells, which were generated by stable transfection of the murine ASC gene into the murine RAW 264.7 macrophage cell line, which is naturally ASC-deficient [2].

• RAW-ASC cells – Murine ASC gene expression

In contrast to the parental cell line, RAW-ASC cells produce and secrete mature IL-1β upon activation of the canonical inflammasome sensors NLRP3 (using Nigericin) and AIM2 (using Poly(dA:dT)), as well as upon activation of the non-canonical inflammasome (using LPS or E. coli outer membrane vesicles). This cell line is a useful tool to study the role of ASC in inflammasome responses and is an alternative to the in vitro differentiation of mouse bone-marrow-derived macrophages. Additionally, it is the parental and control cell line of InvivoGen's RAW-ASC KO-GSDMD cells, for which all inflammasome responses are abrogated.


FEATURES OF RAW-ASC CELLS:

  • Verified expression of the murine ASC gene by Western Blot (Wes™)
  • Validated secretion of mature IL-1β and pyroptosis after canonical and non-canonical inflammasome activation
  • Parental cell line for RAW-ASC KO-GSDMD cells and RAW-ASC KO-CASP11 cells

 

References:

1. Mathur A. et al., 2017. Molecular mechanisms of inflammasome signaling. J. Leuk. Biol. 103:233.
2. Pelerin P. et al., 2008. P2X7 receptor differentially couples to distinct release pathways for IL-1β in mouse macrophage. J. Immunol. 180:7147.

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
Mouse
Tested applications

Inflammasome cellular assays

Cell type
Monocytic
Growth properties
Adherent
Tissue origin
Mouse macrophages
Antibiotic resistance
Blasticidin
Growth medium

Complete DMEM (see TDS)

Mycoplasma-free

Verified using Plasmotest™

Quality control

Each lot is functionally tested and validated.

CONTENTS

Contents

  • Product: 
    RAW-ASC Cells
  • Cat code: 
    raw-asc
  • Quantity: 
    3-7 x 10^6 cells
Includes:
  • 1 ml of Normocin™ (50 mg/ml). Normocin™ is a formulation of three antibiotics active against mycoplasma, bacteria, and fungi.
  • 1 ml of Blasticidin (10 mg/ml)

Shipping & Storage

  • Shipping method:  Dry ice
  • Storage:

    • Liquid nitrogen vapor
    Stability: 20 passages

Details

Inflammasomes are multimeric protein complexes that are crucial for host defense against infection and response to endogenous danger signals. The canonical inflammasome response is driven by aggregation of a sensor (i.e. NLRP3) with the ASC adaptor and pro-caspase-1. Activation of caspase-1 (CASP1) induces the maturation of pro-IL-1β/pro-IL-18 and cleavage of the pore-forming protein gasdermin D (GSDMD), leading to secretion of IL-1β/ 18 and pyroptosis. 

ASC is essential to inflammasome sensors that do not contain a CARD domain, such as NLRP3, AIM2, and Pyrin [1]. This is due to the bipartite composition of ASC, consisting of one PYD and one CARD domain, allowing the recruitment of the CARD-containing pro-caspase-1 to these sensors. Whereas, the NLRP1 and NLRC4 inflammasome sensors have a CARD domain, and can recruit pro caspase-1 either directly or through ASC. However, it has been shown that upon activation of these sensors, in the absence of ASC, the secretion of mature IL-1β and IL-18 is reduced [1]. As the non-canonical inflammasome (i.e. CASP4/5/11) can not directly activate CASP1, they trigger GSDMD-driven release of alarmins and K+ efflux to induce the activation of NLRP3 and CASP1-mediated IL-1β/-18 maturation and secretion. Therefore, the absence of ASC affects the downstream inflammatory signaling from the non-canonical inflammasome also. 

 

1. Mathur A. et al., 2017. Molecular mechanisms of inflammasome signaling. J. Leuk. Biol. 103:233.

DOCUMENTS

Documents

RAW-ASC Cells

Technical Data Sheet

Validation Data Sheet

Safety Data Sheet

Certificate of analysis

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