Label ATTO-488. Maximum absorption 501 nm; maximum fluorescence 523 nm. The fluorescence is excited most efficiently in the 480 – 515 nm range. This label is analogous to the well known dye fluorescein isothiocyanate (FITC) and can be used with filters typically used to detect FITC.
Storage before reconstitution The antibody ships as a lyophilized powder at room temperature. Upon arrival, it should be stored at -20°C.
Reconstitution 50 µl double distilled water (DDW).
Antibody concentration after reconstitution 1 mg/ml.
Storage after reconstitution The reconstituted solution can be stored at 4°C, protected from the light, for up to 1 week. For longer periods, small aliquots should be stored at -20°C. Avoid multiple freezing and thawing. Centrifuge all antibody preparations before use (10000 × g 5 min).
Standard quality control of each lot Western blot analysis (unlabeled antibody, #ANT-007), and immunocytochemistry (labeled antibody).
Applications: fc, ic, if, ih, lci
Immunohistochemistry
Rat DRG sections.
Direct flow cytometry
Rat pheochromocytoma PC12 cells (1:20).
Cell surface detection of p75NTR in live intact rat pheochromocytoma (PC12) cells:
Cell surface detection of p75NTR in live intact human A875 melanoma cells. A. Cells were stained with Anti-p75 NGF Receptor (extracellular)-ATTO Fluor-488 Antibody (#ANT-007-AG), (1:50), (green). B. Live image of the cells. C. Merge of the two images.
Scientific background
The p75 neurotrophin receptor (p75NTR) is a member of the tumor necrosis factor (TNF) receptor superfamily. Like all members of this superfamily, p75NTR is a type I transmembrane protein with tandem cysteine-rich domains in its extracellular portion, as well as an intracellular death domain. p75NTR binds to its ligands as a homodimer but can also form heterodimers with other receptors such as TrkA, TrkB, TrkC, Nogo receptor, and sortilin. The precise multimeric receptor complex formed between p75NTR and the other receptors determine the ligand being recognized (see below) and the biological response to its binding1-3.
As its name implies, p75NTR binds to all the neurotrophins (NTR) (NGF, BDNF, NT-3, and NT-4) with similar nanomolar affinities. Co-expression of p75NTR with the Trk receptors enhances its ability to bind and respond to the specific neurotrophin and induces cell survival. On the other hand, it has recently been demonstrated that p75NTR binds with high affinity to the unprocessed form of NGF (proNGF), probably as a complex with sortilin, leading to cell death by apoptosis1-3. Finally, a multimeric complex of p75NTR and Nogo receptor binds to myelin proteins such as Nogo-66, MAG, and OmgP, resulting in inhibition of axonal growth4.
Application key:
CBE- Cell-based ELISA, FC- Flow cytometry, ICC- Immunocytochemistry, IE- Indirect ELISA, IF- Immunofluorescence, IFC- Indirect flow cytometry, IHC- Immunohistochemistry, IP- Immunoprecipitation, LCI- Live cell imaging, N- Neutralization, WB- Western blot