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Michelle van Geldermalsen

Articles by Michelle van Geldermalsen:

An overview of the Yeast one-hybrid assay

If you are regularly doing ChIP-qPCR, ChIP-RNAseq or luciferase reporter assays to measure protein-DNA interactions, then this article is for you! ChIP experiments can tell you what DNA sequences your protein binds, and luciferase reporter assays predict whether your protein functionally binds a specific promoter to activate transcription – but a yeast one-hybrid (Y1H) assay…

09 Jul 2016 Protein Expression & Analysis

Go Fishing for RNA-Protein Interactions with a Yeast Three-Hybrid Assay

If you’re hoping to reel in a positive interaction between a protein and an RNA sequence, try to catch a winner with a yeast three-hybrid assay. What is yeast three-hybrid (Y3H)? The Y3H system is based on the same principle as a yeast two-hybrid– namely, that the DNA binding domain and the transcription activation domain…

09 Jul 2016 Protein Expression & Analysis

Split Ubiquitin Yeast Two-Hybrid

If you’ve read our article, An Overview of Yeast Two-Hybrid (Y2H) Screening, you’ll know that one major limitation of conventional Y2H is that your protein-protein interaction must occur in the nucleus for the reporter gene to be activated. So what do you do if your protein is a receptor tyrosine kinase? Or a G protein–coupled…

09 Jul 2016 Protein Expression & Analysis

Breakthroughs in Peptide Translocation: Cell Penetrating Peptides

Cell Penetrating Peptides (CPPs) are the Trojan Horse of cell biology – an innocuous peptide sequence with the special ability to carry virtually any cargo across the plasma membrane. If you have a special delivery that you don’t want to get lost in transit, CPPs are for you! CPPs are short peptides (typically 5-30 amino…

09 Jul 2016 Protein Expression & Analysis

An Overview of Yeast Two-Hybrid (Y2H) Screening

If you are cooking up a way to test if two proteins interact, you need a yeast two-hybrid (Y2H) screen in your recipe book. You don’t want your Y2H to turn out half-baked – so check out this guide to Y2H and we’ll help you make sure yours will rise to the occasion! What is…

11 Apr 2015 Protein Expression & Analysis

Proximity Ligation Assay (PLA) for Dummies

It’s a familiar story – molecular biology meets protein science, they get closer and sparks fly. But how exactly does a proximity ligation assay (PLA) work and how do you make sure yours will have a happy ending? What PLA Does PLA allows you to detect individual proteins or individual protein interactor pairs without ectopic…

26 Jan 2015 Protein Expression & Analysis

Insane in the Membrane! PVDF vs. Nitrocellulose – Which One Comes Out on Top?

When it comes to Western blotting, there’s no denying it: Your membrane is a key player. After all it is the physical scaffold that holds your precious samples and it needs to be up to the challenges you throw at it. But depending on your protein’s properties and your downstream detection steps, finding the optimal…

21 Oct 2014 Protein Expression & Analysis

Get your stripping stripes! Find out how to strip and re-blot your Western

Westerns can be tricky and time-consuming, so make the most of your precious membranes and their proteins. Learn how to properly strip off your antibodies and re-probe with another primary antibody. Why you should strip Scientific reasons: To conserve protein samples that are limited or expensive. So that you can analyse the same sample with…

05 Aug 2014 Protein Expression & Analysis

Block, Stock and Barrel – A Guide to Choosing Your Blocking Buffer

Blocking is the essential third wheel in any antibody/antigen relationship. Correct blocking buffer can perfect your antibody’s ability to bind its antigen, while bad blocking can make specific antibody binding near impossible. Don’t let bad blocking be a stumbling block in your Western blot experiments – read on to find out what blocking achieves and…

17 Jun 2014 Protein Expression & Analysis

Equilibrating your way to a perfect Western blot

If you are struggling to optimise your Western blot protocol, one step to consider is the equilibration of your gel and membrane before transfer. Wondering what this step achieves and whether it’s necessary? You’re not alone! I did dozens of Westerns without ever bothering to equilibrate before I realised that it was having a big…

06 May 2014 Protein Expression & Analysis