Application

• For quantitative fluorescent immunoenzymatic assay of NFkB phosphorylation status in cultured cells.

Key Features

• New and improved. Total assay time reduced from the standard 21 hours to 6.5 hours (hands-on time 2.5 hrs).
• Simple and convenient. Cells are directly cultured in 96-well plates. No cell lysis is necessary.
• Accurate and high-throughput. Protein phosphorylation is normalized to total cellular protein in the same well, greatly minimizing well-to-well variations. Can be readily automated as a high-throughput 96-well plate assay for thousands of samples per day.

Method

• Cell-Based ELISA (FL530/585 nm, 360/450 nm)

Samples

• Cells

Species

• Human, mouse

Procedure

• The assay takes 6.5 hrs, hands-on time 2.5 hrs

Size

• 100 tests

Shelf Life

• 6 months

More Details

• Nuclear factor-kappa B (NFkB) is a transcription factor that plays a central role in many physiological processes, e.g. inflammation, tumorigenesis, and apoptosis. NFkB is activated by a wide variety of stimuli, including inflammatory cytokines such as TNF-α. NFkB is a dimer composed of members of the Rel family of proteins: p65/RelA, c-Rel, RelB, NFkB1/p50, and NFkB2/p52. Phosphorylation of p65/RelA at Ser-536 results in decreased nuclear export and enhanced p65/RelA-dependent transcription. BioAssay Systems cell-based ELISA measures phosphorylated p65(S536) (pNFkB) in whole cells and normalizes the signal to the total protein content. This simple and efficient assay eliminates the need for cell lysate preparation and can be used to study NFkB regulation in short-term and long-term assays.
  
  

  Haven’t Found the ELISA Kit for Your Target

  
  We offer two DIY kits: Custom Cell-based ELISA Kit (EFC1M) for mouse antibodies and Custom Cell-based ELISA Kit (EFC2R) for rabbit antibodies. All that is required is a primary antibody. If you prefer, we can develop the assay for you. Please email or call 1-510-782-9988 x 2 to request assay services

No frequently asked questions for this new product. Please check back later.

For more detailed product information and questions, please feel free to email us. Or for more general information regarding our assays, please refer to Technical Support.

Maniam, P., et al. (2015). Regulatory role of GSK3beta in the activation of NF-kB and modulation of cytokine levels in Burkholderia pseudomallei-infected PBMC isolated from streptozotocin-induced diabetic animals. Tropical biomedicine, 32(1), 36-48. Assay: NFKB phosphorylation in rat blood cell (PBMC).

To find more recent publications, please click here.

If you or your labs do not have the equipment or scientists necessary to run this assay, BioAssay Systems can perform the service for you.

– Fast turnaround
– Quality data
– Low cost