Ramos-Blue™ KD-MyD Cells

NF-κB/AP-1 SEAP reporter B lymphocytes with MyD88 knockdown

Ramos-Blue™ KD-MyD (MyD88 knockdown) is a B lymphocyte cell line specifically designed to monitor MyD88-mediated signaling.

Ramos-Blue™ KD-MyD were obtained by stable integration of an NF-κB/AP-1-inducible SEAP (secreted embryonic alkaline phosphate) reporter gene and stable knowndown of the MyD88 gene.

Due to the knockdown of the MyD88 gene, Ramos-Blue™ KD-MyD do not respond or respond weakly to the activation of receptors which signaling is dependent on MyD88, such as TLR7 and TLR9. However, they are still responsive to TLR3 which signals through TRIF independently of MyD88.

When stimulated, they produce SEAP in the supernatant that can be readily monitored using QUANTI-Blue™, a medium that turns purple/blue in the presence of SEAP.

Ramos-Blue™ KD-MyD cells derive from a human Burkitt’s lymphoma which is negative for Epstein Barr virus.

Figures

Cells were incubated with 1 μg/ml poly(I:C) (TLR3), 10 μg/ml CL264 (TLR7), 2 μg/ml ODN2006 (TLR9) and 100 ng/ml TNF-α. After 24h incubation, NF-κB activation was determined using QUANTI-Blue™.

 

 

Specifications

Antibiotic resistance: Hygromycin B and Zeocin®.

Growth Medium: IMDM (Iscove’s Modified Dulbecco’s Medium), 2 mM L-glutamine, 10% v/v heat-inactivated fetal bovine serum (30 min at 56°C), 100 µg/ml Normocin™, Pen-Strep (50 U/ml-50 μg/ml).

Guaranteed mycoplasma-free.

Contents

• 1 vial containing 3-7 x 10⁶ cells
• 1 ml of Hygromycin B Gold (ultrapure hygromycin B; 100 mg/ml)
• 1 ml of Zeocin® (100 mg/ml)
• 1 ml of  Normocin™ (50 mg/ml)
• 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 and some areas in Asia)