LANCE Ultra total NFκB kits are designed for the detection of NFκB (both phosphorylated and non-phosphorylated) in cell lysates using a simple, homogeneous LANCE Ultra TR-FRET assay (no wash steps). This assay is compatible with both adherent and suspension cells, and is intended to be used as a control or normalization assay in conjunction with the LANCE phospho-NFκB assay.
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For research use only. Not for use in diagnostic procedures.
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Please note control lysates are sold separately, catalog number TRF4019S.
LANCE® and LANCE® (Lanthanide chelate excite) Ultra are homogeneous (no wash) TR-FRET (time-resolved fluorescence resonance energy transfer) technologies. One antibody of interest is labeled with a donor fluorophore (a LANCE Europium chelate) and the second antibody is labeled with an acceptor fluorophore [ULight™ dye]. Upon excitation at 320 or 340 nm, energy can be transferred from the donor Europium chelate to the acceptor fluorophore if sufficiently close for FRET (~10 nm). This results in the emission of light at 665 nm. Data are represented as ratiometric (665/615 nm X 10,000).
NFκB (Nuclear Factor kappa B) is a superfamily comprised of p65, RelB, c-Rel, p50/p105, and p52/p100. The combinations of these as either homo- or heterodimers help to regulate the immune system in response to infection or cellular stress. The classical NFκB pathway involves p65 and p50 activation and in stimulation of cytokines or activation of TLR. Phosphorylation at S536 has been shown to enhance transcriptional activity in the nucleus of cytokines as part of immune response. Several auto-immune disorders have been traced back to the deregulation of the NFκB pathway.
|Assay Target Class||Protein|
|Detection Method||Time-Resolved Fluorescence (TRF), TR-FRET|
|Product Brand Name||LANCE Ultra|
|Shipping Condition||Blue Ice|
|Unit Size||10,000 Assay Points|
This application note describes a comparison of two TR-FRET based technologies for measuring phospho-ERK