Targeting Tools: mu p75-SAP

The cover article this quarter demonstrates usage of mu p75-SAP (Cat. #IT-16) to study delirium. To create this toxin, we affinity-purified the rabbit polyclonal to the low affinity neurotrophin receptor, p75, (Cat. #AB-N01AP) with the immunogen bound to a solid support, and then conjugated the affinity-purified antibody to saporin (Cat. #PR-01). As can be seen in the cytotoxicity assay below, mu p75-SAP has an EC50 in the nanomolar range. This high potency translates to smaller amounts used for elimination of p75-positive neurons in the mouse brain which results in a greater index of efficacy and lesser non-specific cytotoxicity.

The mu p75-SAP kit includes, in addition to the immunotoxin, equal aliquots of saporin (Cat. #PR-01), the affinity-purified rabbit polyclonal antibody (Cat. #AB-N01AP), and the control immunotoxin, Rabbit-IgG-SAP (Cat. #IT-35).

Also available are fluorescent conjugates of AB-N01AP: Cy3-labeled Anti-murine NGFr (Cat. #AB-N01APFL3), and Cy5-labeled Anti-murine NGFr (Cat. #AB-N01APFL5).

mu p75-SAP
Micrographs of brain coronal sections of mice treated with PBS or mu p75-SAP. In lesioned mice, the number of ChAT-positive neurons is dramatically reduced in the medial septum (MS) and the diagonal band of Broca (DBB) (a,c) and in the nucleus basalis (NB) (b,d). AChE staining is massively depleted in the cortical mantle (e,g) and the hippocampus (f,h), but not in the thalamus (Th). Scale bar = 200 µm.
(Fig 1 from Moreau et al. Targeting Trends, 9(2): pp. 1,6, 2008.)
NG6 cells are plated at 1000 cells/well and incubated overnight. Saporin and mu p75-SAP (conjugate of the affinity-purified rabbit polyclonal to mouse NGFr and saporin) are added in 10-µl volumes and the plates are incubated 72 hours. PMS/MTS developing reagent is added and the plates are incubated 1-2 hours, then read at 490 nm.
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