Image-iT™ Low Density Lipoprotein Uptake Kit, pHrodo™ Red

The Image-iT Low Density Lipoprotein Uptake Kit - pHrodo Red includes pHrodo Red dye-labeled LDL (also available separately), unlabeled LDL, heparin, BSA, and assay buffer. The kit was designed to specifically detect binding and uptake of LDL through the LDL receptor internalization pathway, allowing maximum control and flexibility in the experimental design. It can be used with fluorescence microscopy, flow cytometry, cell sorting, high-throughput analysis, and high content analysis.

• pHrodo Red-LDL is detectable with RFP/TRITC filter set (absorption peak: ∼560 nm, emission peak: ∼585 nm)
• pHrodo Red-LDL is mutiplexable with green, deep red, violet, and blue emission dyes, such as GFP, DAPI, Hoechst, HCS LipidTOX, and LysoTracker Deep Red
• No-wash protocol is possible, allowing dynamic measurement of LDL uptake and trafficking

pHrodo Red- LDL is dimly fluorescent at neutral pH outside of cells, but fluoresces brightly after endocytosis. This feature allows for a no-wash protocol when using pHrodo Red-LDL, enabling kinetic measurement of LDL uptake, endosomal maturation, and trafficking in cells.

Two controls are included in the kit: unlabeled LDL as an optional pretreatment to occlude cell surface receptors in advance of probing with the labeled construct, and heparin which will chelate the LDL in the solution phase outside of the cells, preventing binding and uptake. When cells are labeled in the absence of these pretreatment controls, a fluorescent signal indicates specific LDL receptor-mediated endocytosis.