You obtain the most effective and cleanest IPs of your protein when using ChromoTek’s Nano-Traps (derived from alpaca single domainantibodies). You will see the best protein detetions in WB when applying our highly sensitive monoclonal antibodies.
See the advantages of ChromoTek’s RFP-Trap in ChIP applications as demonstrated by Dr. Caroline Rivers, Laboratory for Integrative Neuroscience & Endocrinology, University of Bristol. In absence of suitable antibodies for an inducible hormone receptor, mCherry has been fused to the N-terminus of the receptor. In order to effectively pull down the mCherry fusion protein DNA complex, ChromoTek's RFP-Trap was used.
The ChromoTek rat monoclonal anti-RFP antibody 5F8 has been cited for the use in immunofluorescence (IF) and immunohistochemistry (IHC) experiments by 91 peer reviewed articles. It is thus the anti-RFP antibody referenced most often at the database citeAB. This top rank is the result of its performance and quality.
Everybody really wants to use a detection probe that does not influence its target. For measurement of actin and actin-related functions there are a number of actin visualization probes and techniques available that researchers can select. Hence, one should only apply such actin visualization probes, which do not alter actin dynamics. How do the actin probes for visualization perform? Which probes do interfere with function and which probes do not? Robert Grosse and his team from the Institute of Pharmacology at the Biochemical-Pharmacological Center (BPC), University of Marburg, Germany took the effort and systematically looked into detection technologies of actin filaments in order to avoid potential pitfalls. In the recently published mini review/poster “Actin Visualization at a Glance” (Melak et al. 2017) they have discussed the pros and cons of current probes to visualize actin filaments, i.e. Phalloidin, LifeAct, Utrophin, F-tractin, SiR-actin, GFP-actin, tagged actin, and Actin Chromobody® (see table 1 and figure 1 below).
Although traditional IgG antibodies are often used for immunoprecipitation and protein interaction analysis, GFP-binding protein (GBP, ChromoTek gt-250) and other ChromoTek VHHs (http://www.chromotek.com/about-us/the-alpaca-antibody-advantage/) easily outcompete those under challenging conditions such as elevated temperatures, varying pH or high denaturant concentrations. In fact, GBP tightly binds GFP-fusion proteins even in 8 M urea at 51 °C, which sets a new limit in protein complex stability, virtually unrivalled by any other capture molecule protein tag pair.
The progression of epithelial cancers into metastatic stages is triggered by a cellular process called “epithelial-mesenchymal transition” (EMT). In response to cytokines, tumor cells shut down the expression of epithelial makers including occludin or E-cadherin and gain mesenchymal markers such as N-cadherin or vimentin. Such cells are losing their cell-cell contacts and apical-basal polarity and become highly migratory and invasive. Considering the importance of this process for cancer progression and formation of metastasis, reliable cellular EMT biomarker and versatile detection systems are necessary to develop novel anti-metastatic therapies.
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Please use this email address or, alternatively place your order via our online portal www.chromotek.com/webshop to benefit from improved delivery times.
On a personal note, we – ChromoTek - like to state a few words on the quality of our camelid VHH or heavy chain antibodies (also known as “nanobodies”).
Topics: Antibody Quality
Pull-down of proteins can be difficult, particularly when they are expressed at low levels. Here we present 5 tips, which will considerably improve your IP results.
- Protein concentration matters
The higher the protein concentration, the higher the IP yield. Try to use a concentration as high as possible.It makes a significant difference if the concentration of your protein during IP is 1 nM (low protein expression level), about 50 nM (intracellular endogenous protein expression level, will be diluted by buffer for IP) or 1000 nM (high protein concentration).
Cell cycle and cell proliferation are tightly controlled cellular processes, and their deregulation is a hallmark of cancer. To enable non-invasive analysis of cell cycle in living cells, ChromoTek developed a Cell Cycle Chromobody. The Cell Cycle Chromobody (CCC) consists of a binding domain of a heavy-chain antibody highly specific against human proliferating cell nuclear antigen (PCNA), which is genetically fused to a fluorescent protein.