Atlas Antibodies Blog

This month we celebrate the first anniversary of our blog. It has been an incredible journey. If you have missed our blog posts, read on. We have interviewed scientists, discussed the importance of antibodies validation, the human protein atlas project and we gave you advice on how to succeed with your immunohistochemistry.

Today we meet with Dr. Jan Mulder, group leader of the Human Protein Atlas (HPA) brain profiling group at Karolinska Institutet, Stockholm, Sweden, and the director of the Brain Atlas, a brand-new addition to the (HPA) project. His research interest is to identify proteins involved in brain development, normal brain physiology and pathophysiology of brain disorders by using an antibody-based approach combined with multiplex fluorescence immunohistochemistry.

We asked Jan 10 questions about the past, the present, and the future of the Brain Atlas.

Applied biological or biomedical research aims to reach a thorough understanding of the physiology of organisms and to further development of disease therapeutics. Some scientific discoveries have narrow applicability or scope and are sometimes serendipitous or even entirely unexpected. Other have wider applicability and transferability having proved enormously useful over a wide range of domains. Among the latter are research antibodies. Read more to discover how they help progressing science.

Washing- and staining solutions, blocking agents, detection and mounting solutions. Successful immunohistochemical staining requires reliable, high-quality reagents to fixate, to wash and permeabilize the tissue. But what is the rationale behind the fact that certain buffers or storage conditions are better to use than other, or is it all just empiric?

Today we meet with Dr. Cecilia Lindskog, group leader at Uppsala University in Sweden, and the site director of the Tissue Atlas part of the Human Protein Atlas (HPA). Her research is focused on protein science, understanding the biology and functions of human proteins expressed in different organs, and the underlying mechanisms leading to cancer and other diseases. We asked her 10 questions about the past, the present, and the future of the Tissue Atlas.

As a rule, the complex biology and spatial organization of tissues and solid tumors poses a scientific and diagnostic challenge that is not sufficiently well addressed by standard immunohistochemistry (IHC) procedures. How come this is the case? The answer is that single IHC staining provides data on only one marker at a time. So, how do you stain multiple protein in the same cell or tissue?

In this blog, we would like to tell you about that time when researchers interested in a new therapeutic target for patients with type 2 diabetes, knocked at our door asking for help, and how we used the Tissue Atlas, part of the Human Protein Atlas project, to answer their questions. But let’s start from the beginning.

Today we meet with Dr. Kristian Moller, Principal Scientist at Altas Antibodies, to ask him 10 questions about past, present, and future of multiplex immunohistochemistry.

Today we meet with Dr. Caroline Kampf to ask her 10 questions about past, present, and future of immunohistochemistry (IHC). 

How many samples? How much money and time have you wasted trying to unmask a protein on your IHC samples? Tissue fixation in immunohistochemistry often negatively impacts on your immunostaining by masking the epitope of interest. Read on to learn about the most common antigen retrieval methods used to restore epitope-antibody binding.

Would you print your childhood best and precious memories on a low-quality photographic film? I wouldn’t. However, just like me, you probably have hundreds of old faded yellow photos abandoned in a drawer. Images quality relies on high-quality printing films…and so does your immunohistochemical staining. Tissue handling and tissue processing determine the quality of your IHC results. You want high resolution?  Let’s understand how to achieve it.

 “I don’t know why I am doing this. I can’t handle it!”  You thought you knew how to recognize and solve your IHC artifacts but after reading the troubleshooting guide the answers seem so far away. Long lists of tips. Examples too difficult to understand. Not-so-friendly images. Feeling overwhelmed? You are not alone. Although sometimes troubleshooting seems unbearable, there is no need to worry! Troubleshooting is nothing more than the logical elimination of variables.

After a few intense days of meticulous tissue washes and antibody incubations my staining is finally ready. I am staring at it down the microscope. I enjoy what I see: a dark brown color exactly where I expect to see it! Am I good or what? I give myself a figurative pat on the back and an imaginary high five. Excited and proud, I call my supervisor who glares intently at it and then firmly, and rather surprisingly, asks me to re-run the staining. What went wrong? What did I miss? I must get to the bottom of this!

How many decisions do you think you make every day? Would you be surprised if I tell you that an adult person makes about 25,000 more or less conscious decisions every day? It’s a fact. Consciously or unconsciously, we spend every day of our lives making decisions: where to go, what to wear, what to eat, how to spend our time, etc. But how much time do you spend making a decision for the right control for your primary antibody? And, most importantly, did you make the right choice? How can you tell a good decision from a bad one?

Headphones on, I am listening to my favorite music and staring at the lab bench in front of me feeling excited to start the immunohistochemistry experiment of the day. Pipette, timer, wells, vials, brush, buffers. Protocol taped on the shelf. Everything is ready. I feel like a DJ at the console, and I find myself thinking: what makes a good DJ?  A DJ does not just play one song after the other in a random sequence. He has everything under control, he plans and underpins, every minute of a performance to varying degrees. So, how do I get started when planning a new set for a forthcoming immunohistochemistry experiment? Read on to find out how to control your results and prepare for a successful ’gig’ in the lab.