The immunotherapy field is making great progress in aiding many disease treatments. The hope is that one day, personalized immunotherapy can be used so each patient can use their own immune system to overcome their specific disease affliction. QuickSwitch is a tool for creating custom MHC class I tetramers quickly and easily.
The process of autophagy was first recognized in the late 1950s and it was thought of as bulk “junk” removal. Then, dedicated scientists did a little more digging and found there is an amazing methodology to the “junk” removal. It was discovered that there is rhyme and reason for how a cell decides if and when its components should undergo degradation. Autophagy is rather specific and aids in cell survival by making sure the cell has essential components during times of flux. Autophagy can be selective (i.e. mitophagy) or non-selective (i.e. starvation-induced). Using this method, the cell can have non-essential components recycled or send them to onto autophagolysosomes to be degraded. It is more efficient for a cell to be able to recycle as much as it can, rather than to wait for new proteins to come into the picture. Autophagy is also crucial for clearing out “junk” such as misfolded or aggregated proteins. Research in autophagy has helped provide a better understanding for how a cell is able to survive even under poor conditions.
What is mitophagy? It is when damaged mitochondria are removed from the cell by autophagy. The damaged mitochondria end up in lysosomes for their final disposal. This whole process is to maintain and assure proper cellular function. The importance of this biological process is that it has been implicated in disease states such as cancer and Parkinson’s disease.
Have you seen comparisons of multimer technology claiming to be superior to tetramers? Be sure to ask which tetramers were used in the comparison! MBL tetramers have a clear advantage over academic tetramers and other commercial MHC multimer products, not only due to the reliability and high quality for every lot produced, but also due to the proprietary alpha-3 mutation. This mutation, engineered into the heavy chain of all of our human and macaque class I tetramers, helps decrease non-specific binding, leading to enhanced specificity. Check out the images below to see the details of this important technology!
MBL’s CoralHue fluorescent product series is composed of many proteins to make your life "brighter"! We offer expression vectors and cloning vectors. How do you know which vector to use? Learn more about how to choose which vector is right for you.
When you start working in a new lab or working on a new research project, one of the first things to do is look up publications and learn about the field and what has been accomplished. This will help determine the experiments you will run. It gives you a foundation to build on. But how do you read a publication? How do you know it’s worth your while to muster through? Some publications are miles long with a million figures! EEKS! Don’t be frightened. Keep reading to find out how to tackle a complicated publication and not go crazy.
You have an RNA of interest aka your favorite RNA. The question is: which RBPs interact with it? Who are these RBPs and what are their intentions with your precious RNA?
How do you quantify newly synthesized RNA? There are several ways to detect synthesized RNA, but these methods can be inaccurate due to toxicity.
What happens when you use a black well plate for a fluorometric ELISA assay reading? What happens when you use a clear well plate for a colorimetric ELISA assay reading? Well the short answer is that you made the right plate decision!
Topics: Kits and Assays
You’re sitting at your flow cytometer, staring at your CD8 x tetramer plot in anticipation of a tiny but meaningful group of dots appearing in the upper right quadrant. You hold your breath, click “acquire,” and watch… Wait, what? What’s this ugly diagonal?? How come ALL the CD8 positives are dual positive??! Hey, this is my control mouse! That one should be completely tetramer negative! Yep, I’ve been there too. Classic case of using a CD8 antibody that doesn’t play nice with tetramers.
Figure: Schematic representation of feedback loop involving CLOCK-BMAL1 complex and CRY and PER proteins involved in creating circadian rhythms.
Tiredness and frustration often mount after long nights in the lab. Why can’t our bodies adjust and allow us to maintain our energy through the night? The answer lies in our circadian rhythms. Circadian rhythms are the daily cycle of biological processes such as wakefulness and metabolism1. Many proteins are involved in regulating our circadian rhythms including CRY proteins. This blog post will catch you up on these proteins and their vital role in regulating circadian rhythms.
What does Autophagy Flux even mean? It’s not in the Urban dictionary. Dear reader, you have come to the right place to learn a little more about autophagy flux and how our Autophagy Watch kit can help you.
Figure: Schematic representation of S100 proteins in the MAP kinase and NF-kappaB pathways.
S100 proteins are a family of proteins known for their role in a variety of cellular pathways and diseases. This blog post will catch you up on the latest research on these important proteins.
Figure: Schematic representation of PCSK-cIAP1 Binding Assay Kit which can be used to screen inhibitors of PCSK9-cIAP1 interaction during apoptosis.
As discussed in two prior blog posts, PCSK9 has primarily been studied for its role in regulating cholesterol levels and as a target for treatments to lower cholesterol. However, PCSK9 has been found to be involved in many other important pathways.
This blog post will catch you up on the latest research into the diverse functions of PCSK9.
Ever wonder how cholesterol levels are regulated? One of the major regulators is a molecule called proprotein convertase subtilisin/kexin type 9 or PCSK9 for short! PCSK9 has been implicated in causing hypercholesterolemia (high cholesterol), and diseases such as coronary heart disease1.
This blog post will get you caught up on cholesterol and research on PCSK9's role in cholesterol regulation.
You have been given the mission to start a new project. A tiny tube is put in your hands and you’re told it contains a potentially interesting protein and your job is to characterize it. After you wipe nervous sweat from your brow, you get down to business. Step one- how to isolate your new buddy? Immunoprecipitation can help with this question! You determine which epitope tag is on your protein and go to mblintl.com to see what you can buy to help this new adventure. You see there are antibodies conjugated to magnetic beads, which help protein pull downs from cell lysates. It's easy, it's fast. Hmmmm- promising.
Topics: Epitope Tag
Given the prominent role of Fas in many important cell functions (7 Facts to Catch You Up to Speed on Fas), Fas has been implicated in a variety of disease states such as cancer. Here are some major findings from recent research about the role of Fas in cancer:
Fas antigen is a receptor protein that has been linked to many diseases such as cancer1 and autoimmune diseases2. Here are seven findings from research on this protein to catch you up to speed on Fas:
In the previous Tetramer Tips blog, I suggested ways for you to be a negative Control Freak. Now it’s time to think positive! A positive control for a tetramer is a sample that contains cells expressing the specific T cell receptor of interest, i.e. has the exact specificity of the tetramer. Okay, brace yourselves; I’m going to be frank here. Having a positive control for tetramer experiments is often a quest for the Holy Grail. In many (most, in fact) cases, you will not have access to a positive control, unless one of your experimental samples happens to show a positive result. In an experiment where no positive events are seen and no positive control was used, you cannot necessarily conclude that the donor/patient/mouse is negative for that T cell specificity, because, heck, maybe you got distracted while pipetting and forgot to add the tetramer to your staining cocktail! A tetramer experiment with no positive control and no positives in the experimentals is, therefore, uninterpretable. This is the hard truth we must face.
What are LC3 isoforms all about? What do they do? How are they different? How are they involved in autophagy?
Antibodies are produced in abundance by the human immune system. The white blood cells are responsible for their manufacture, including lymphocytes (T-cells and B-cells) and antigen presenting cells. Their job is to produce a coordinated response to foreign substances (antigens) as well as to destroy harmful pathogens such as bacteria and viruses. The human body contains vast numbers and varieties of normal and essential human proteins called autoantigens. In some situations the body’s immune system makes an error and recognizes the normally occurring autoantigens as “foreign,” binding to them in order to destroy and eliminate them from the body. A specific subset of antibodies that attack normal proteins found in the nucleus of the cell are called, not surprisingly, antinuclear antibodies, or “ANAs” for short. These misguided antibodies direct their attack against the cell’s nucleic acids and sometimes their associated proteins which are found in or near the nucleus of the cell. Elevated levels of ANAs have been associated with various rheumatic diseases including systemic lupus erythematosus (SLE), rheumatoid arthritis, Sjögren's syndrome, scleroderma, polymyositis, dermatomyositis and various other systemic autoimmune diseases.
While being a Control Freak may not be the best choice for living in spiritual harmony, it is a downright asset in flow cytometry research! Controls not only help you set up an experiment to get a clear or “true” answer, they can also help you troubleshoot what may have gone wrong when your data just doesn’t look quite right, so that your next attempt will turn out better. In flow cytometry, controls are critical to help determine “real” events from artifacts. So, what should you use as controls in tetramer experiments?
Mitophagy is the selective degradation of old or depolarized mitochondria by autophagy, mitophogy contributes to maintaining a healthy population of mitochondria. Since damaged mitochondria leads to collapsed cell homeostasis, mitophagy is believed to protect against diseases related to mitochondrial dysfunction such as neurodegenerative disorders, most importantly Parkinson's Disease.
Inflammation is regulated by small glycoproteins called cytokines. Cytokines are cell signaling proteins in the immune response pathway1. Il-18 is a cytokine that primarily facilitates Th1-type immunoreactions by acting on T cells.
Antibodies are wonderful. They can act as little detectives to help you determine if the protein you’re interested in is involved in a certain process or help characterize the function. But what if your favorite protein has two forms? Is your antibody sophisticated enough to be able to detect one form over the other? Does this knowledge help you? How? IL-18 is one such protein that has two forms. It is present when it is inactive and present when it is active. So what is known about IL-18 and its function and the two forms it can exist in?