In the Sept 14th 2017 publication in Science, the challenges and pitfalls associated with the use of antibodies were highlighted. This is not a new revelation, as the problems surrounding the use and validation of antibodies have been discussed in the past (see articles bottom of the page). There are a limited number of commercial antibody manufacturers that develop and validate their own antibodies, with articles citing that these companies operate more like department stores. As a consequence, the IHC user would buy from one company and have to perform their own independent validation, costing lots of money, time and samples, often resulting in questionable results. In absence of a quick and easy antibody validation method, orthogonal validation like RNAscope technology can provide an initial quick and easy way to validate antibody results.
Distribution of the c-MYC gene product in colorectal neoplasia (Histopathology 2016), Baker et al. use RNAscope™ Assay to detect MYC mRNA in clinically relevant samples, and thereby determine the reliability of MYC targeting antibodies.
The authors performed IHC on human FFPE normal colon (n = 15), hyperplastic polyps (n = 4) and neoplastic colon samples (n = 55) using the N-terminally directed antibody Y69, the C-terminal antibody 9E10, and compared with RNAscope™ Assay in serial sections.
They found the localisation of MYC mRNA correlated well with the protein distribution determined by the N-terminally directed antibody Y69 while the 'gold standard' antibody 9E10 often showed a reciprocal pattern of expression.
They conclude that "the observed discrepancy between the staining patterns suggests that the significance of 9E10 in immunohistochemical staining is currently uncertain, and therefore should be interpreted with caution"
Easy single-day RNAscope Assay provides rapid validation.
RNAscope® Assay is sensitive, specific, reliable and cited in nearly 1000 publications.
Designed for virtually any target, in any species.
Enables a quick comparison of RNA and protein expression (using IHC antibodies).
RNA-based detection, RNAscope assay is truly an orthogonal method that both validates and can provide additional insights for gene expression.
Widely used by industry; protocols available
Expert team with over five years of experience in RNAscope™ ISH
Each project is individually tailored and managed by our team of scientists, specialists, board-certified pathologists, and data experts.
Get your RNAscope results in 5 to 6 weeks for only $/€5000!
*This promotion is available only to life science professionals 21 years or older in the US (excluding Puerto Rico), Canada, Albania, Andorra, Armenia, Austria, Belarus, Belgium, Bosnia and Herzegovina, Bulgaria, Croatia, Cyprus, Czech Republic, Denmark, Estonia, Finland, France, Georgia, Germany, Greece, Hungary, Iceland, Ireland, Italy, Kazakhstan, Kosovo, Latvia, Lithuania, Luxembourg, Macedonia, Malta, Moldova, Monaco, Montenegro, Netherlands, Norway, Poland, Portugal, Romania, Serbia, Slovakia, Slovenia, Spain, Sweden, Switzerland, Turkey, Ukraine and United Kingdom. Discount will apply to orders and samples received by Advanced Cell Diagnostics no later than Dec 1, 2017. This offer is valid for any Pharma Assay Services order with a minimum of 2 targets per order. The validation will be performed using up to 4 samples provided by the customer. RNAscope data provided for this antibody validation consists of stained slides, scanned images, semi-quantitative scoring and a brief report. Contact your account representative if the validation needs to be performed on more than 4 samples. Cannot be combined with other discounts or promotions. Other restrictions may apply. Advanced Cell Diagnostics has the final discretion in determining the applicability of all the promotions.
