Frequently Asked Questions

 

What is MolBoolean™?

MolBoolean™ is a novel in situ proximity technology developed by Atlas Antibodies that enables the simultaneous detection of both free and interacting fractions for two protein targets in cells and tissue. ​

What are the benefits of using MolBoolean™ compared to traditional methods for studying protein-protein interactions?

MolBoolean™ offers several advantages over traditional methods, including the ability to detect both interacting and non-interacting fractions of proteins, normalization of data to account for changes in target protein levels, and a consistent molecular process that ensures uniform signal efficiencies.

How does MolBoolean™ overcome the limitations of in situ PLA?

MolBoolean™ overcomes the limitations of in situ PLA by providing more comprehensive information on protein interactions. It can detect both interacting and non-interacting fractions of proteins, allowing for a complete spatial quantitative analysis. Additionally, MolBoolean™ offers normalization of data to account for dynamic changes in protein complex formation, addressing a key gap in data interpretation.

How does MolBoolean™ normalize the data and why is this important?

MolBoolean™ normalizes the number of protein interactions to the total number of target proteins. This is crucial because target protein levels can be influenced by various factors such as cell treatments or disease states. By providing a normalized measure, MolBoolean™ allows for more accurate comparisons between samples and enhances the reliability of the results.

Can MolBoolean™ detect low abundant proteins?

Yes, MolBoolean™ can detect low abundant proteins. It utilizes rolling circle amplification (RCA), which can increase the fluorescence signal by 1000-fold, allowing for the detection and quantification of low abundant proteins.

How does MolBoolean™ differentiate between interacting and non-interacting proteins?

MolBoolean™ relies on dual target recognition with proximity probes and generates rolling circle amplification products. By performing a series of proprietary molecular steps, MolBoolean™ can differentiate between amplified products stemming from individual versus interacting proteins. This differentiation is indicated by the binding of one or two separate fluorescent detection reporters.

Can MolBoolean™ be used with any primary antibodies?

Yes, MolBoolean™ is a universal kit that can be used with your choice of primary antibodies (one raised in mouse and the other in rabbit).This adaptability makes it suitable for different research needs and allows you to use antibodies that are specific to your proteins of interest.

Is the specificity of the primary antibodies used in MolBoolean™ important?

Yes, the specificity of the primary antibodies used in MolBoolean™ is important for the accuracy of the results. You need to validate the primary antibodies to ensure that they specifically target the proteins of interest. At Atlas Antibodies we take great care to validate our antibodies. We apply Enhanced Validation to ensure antibodies specificity.

Has MolBoolean™ been validated in both cells and tissue?

Yes, MolBoolean™ has been validated in a number of cell lines and tissues, such as MCF7 cells, human kidney, and human colon.

What is the volume of the kit and how many assays does it cover?

The volume of the MolBoolean™ kit is 4.8 ml. The kit covers approximately 120 assays in cells (using 40 μl/assay), and 60 assays in tissue (using 80 μl/assay).

Are there some examples of peer-reviewed applications of MolBoolean™?

Yes, there are.

Raykova et al. (2022) A method for Boolean analysis of protein interactions at a molecular level. Nat Commun. 2022 Aug 13;13(1):4755. Erratum in: Nat Commun. 2023 Sep 6;14(1):5450. PMID: 35963857

Kotliar et al. (2023) Expanding the GPCR RAMP interactome. bioRxiv [Preprint]. 2023 Nov 23:2023.11.22.568247. PMID: 38045268;

Rivas-Santisteban et al. (2023) Boolean analysis shows a high proportion of dopamine D2 receptors interacting with adenosine A2A receptors in striatal medium spiny neurons of mouse and non-human primate models of Parkinson’s disease. Neurobiol Dis. 2023 Nov;188:106341. PMID: 37918757.