AURION IMMUNOGOLD REAGENTS EDITION VI APPLICATION EXAMPLES


Routine paraffin section of Hodgkin lymphoma stained for CD 15. Reed-Sternberg cells show positive staining in the cytoplasm. PRODUCTS USED: • Mouse monoclonal CD 15 • GAM lgG UltraSmall • Aurion R-Gent SE-LM Courtesy of H. Stoop, Laboratory of Pathology and Anatomy, Dordrecht, The Netherlands.


TECHNICAL TIPS...


How to control background in a nutshell


Background Is controlled by 3 independent steps


1. Low Molecular Weight Block (before protein block)


Purpose: to inactivate residual fixative e.g. aldehydes, using


• amino acids such as glycin or lysin, or


Immunogold labeling of Middle East Respiratory Syndrome-coronavirus (MERS-CoV).


MERS virus is grown in VERO cells via a grid cell culture technique. Primary antibody is a camel anti MERS serum which reacts with the glycoprotein spikes of the virus. Detection with Protein A 10nm. After incubation with the gold reagent specimens are fixed in glutaraldehyde and negative stained with Nano-W.


Courtesy of Sandra Crameri, Australian Animal Health laboratory CSIRO, Geelong, Australia


• aldehyde inactivating compounds such as NaBH4 and NH2OH


2. High Molecular Weight Protein Block (before immunolabeling) Purpose: to prevent stickiness to hydrophobic areas and domains with excessive positive charges based on multiple point interactions (high affinity protein binding capacity), using


Regular Light Microscopy • albumin • normal serum


3. Incubation and Wash solution (during immunolabelling) Purpose: to eliminate aspecific binding of immunoconjugates based on hydrophilic interactions (“oligo” point interactions) by competition, using • acetylated albumin (AURION BSA-c™) in the incubation buffer


Please refer to our Newsletter 1 and Newsflyer 1 on our website for more information on the subject of background. You can find incubation protocols in "The Aurion method" section.


Immunogold silver staining of alpha-amylase on Lowicryl HM20 section of rat pancreas. • Goat-anti-Rabbit,15nm


Combination regular light microscopy and epi-polarization microscopy


Immunogold silver staining of alpha-amylase on Lowicryl HM20 section of rat pancreas. • Goat-anti-Rabbit


• Ultra Small Silver Enhanced Epi-polarization microscopy


IGSS of tubulin on coverslip culture of PtK2 cells Courtesy of Peter van de Plas, Aurion, Wageningen, The Netherlands.


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