MolBoolean Starter kit

MolBoolean(TM) Mouse/Rabbit is a kit for in situ protein proximity analysis in tissue and cells. The MolBoolean assay utilizes a proprietary oligonucleotide setup that enables the simultaneous detection of both free and interacting (~40 nm proximity) fractions for two proteins of interest (protein A, protein B and interaction proteins AB). MolBoolean(TM) Mouse/Rabbit should be performed with the user's primary antibodies of choice (raised in mouse and rabbit). MolBoolean relies on anti-mouse and anti-rabbit secondary proximity probes and rolling circle amplification (RCA) as a mean to amplify signal. A series of molecular steps, performed in the MolBoolean assay, incorporates information into the amplified products indicating antibody target engagement of individual versus interacting proteins. This information is converted to fluorescent signals by the binding of detection reporters, allowing visualization on a conventional fluorescent microscope. An image analysis software is provided to segment and differentiate fluorescent signals, enabling the relative quantification of free versus interacting fractions for the two analyzed protein targets in the sample. Included in the kit: Antibody proximity probes, oligonucleotides, enzymes, fluorescent detection reporters. Not included in the kit: Primary antibodies. Target species: Mouse, Rabbit. Product image descriptions: (1) MolBoolean analysis of the interaction between E-cadherin (cat. AMAb90862, magenta) and beta-catenin (cat. HPA029159, green) in MCF7 cells, showing the relative quantification of free versus interacting protein fractions, indicated by the detection of rolling circle products (RCPs) in either one or two fluorescent channels. Data is normalized to total target protein levels (total RCPs). (2) MolBoolean analysis of the interaction between ACE2 (cat. AMAb91259, magenta) and TMPRSS2 (cat. HPA035787, green) in human kidney, showing the relative quantification of free versus interacting protein fractions, indicated by the detection of rolling circle products (RCPs) in either one or two fluorescent channels. Data is normalized to total target protein levels (total RCPs).