Thiol–ene immobilisation of carbohydrates onto glass slides as a simple alternative to gold–thiol monolayers, amines or lipid binding

2015 ◽  
Vol 3 (1) ◽  
pp. 175-181 ◽  
Author(s):  
Caroline I. Biggs ◽  
Steve Edmondson ◽  
Matthew I. Gibson

Carbohydrate arrays are a vital tool in studying infection, probing the mechanisms of bacterial, viral and toxin adhesion and the development of new treatments, by mimicking the structure of the glycocalyx.

2006 ◽  
Vol 6 (2) ◽  
pp. 271-274 ◽  
Author(s):  
Jaeyoung Shin ◽  
Minho Cho ◽  
June Won Hyun ◽  
Jaeho Pyee ◽  
Heonyong Park

ChemBioChem ◽  
2005 ◽  
Vol 6 (6) ◽  
pp. 1007-1015 ◽  
Author(s):  
Moritz B. Biskup ◽  
Jan U. Müller ◽  
Ralf Weingart ◽  
Richard R. Schmidt

2003 ◽  
Vol 51 (8) ◽  
pp. 995-1003 ◽  
Author(s):  
Hidetoshi Ino

Antigen retrieval (AR) is frequently required for successful immunohistochemistry (IHC) in archival formalin-fixed, paraffin-embedded tissue sections. Although AR by heating is most generally used, the majority of existing methods are useful only for paraffin-embedded sections. This article describes a simple alternative method for AR that can be used for aldehyde-fixed frozen sections. After fixation in paraformaldehyde, tissue blocks were heated in retrieval solutions and then frozen with dry ice. The optimal temperatures for heating were 90C and above, and the optimal retrieval solutions were distilled water and 10 mM sodium citrate, pH 6.0. Sections were cut with a cryostat and mounted on poly-L-lysine-coated glass slides. After the sections dried, routine IHC was performed. Alternatively, free-floating sections were used. This method not only greatly enhanced the immunoreactivity for a wide range of antigens, especially for nuclear proteins, but also effectively lowered the background staining in some cases. I examined the staining of 14 antibodies using sections of mouse brain and rat testis. The heating process was essential for five antibodies, improved immunoreactivity for seven antibodies, and provided no change for two antibodies.


Author(s):  
K. Chien ◽  
R. Van de Velde ◽  
I.P. Shintaku ◽  
A.F. Sassoon

Immunoelectron microscopy of neoplastic lymphoma cells is valuable for precise localization of surface antigens and identification of cell types. We have developed a new approach in which the immunohistochemical staining can be evaluated prior to embedding for EM and desired area subsequently selected for ultrathin sectioning.A freshly prepared lymphoma cell suspension is spun onto polylysine hydrobromide- coated glass slides by cytocentrifugation and immediately fixed without air drying in polylysine paraformaldehyde (PLP) fixative. After rinsing in PBS, slides are stained by a 3-step immunoperoxidase method. Cell monolayer is then fixed in buffered 3% glutaraldehyde prior to DAB reaction. After the DAB reaction step, wet monolayers can be examined under LM for presence of brown reaction product and selected monolayers then processed by routine methods for EM and embedded with the Chien Re-embedding Mold. After the polymerization, the epoxy blocks are easily separated from the glass slides by heatingon a 100°C hot plate for 20 seconds.


Author(s):  
Robyn Rufner ◽  
Gerhard W. Hacker ◽  
Michele Forte ◽  
Nancyleigh E. Carson ◽  
Cristina Xenachis ◽  
...  

The use of immunogold-silver staining (IGSS) to enhance label penetration and Localization for immunocytochemistry or in situ hybridization utilizing a variety of metallic salts has been documented. In this morphological study, the effects of silver acetate, silver lactate and silver nitrate were evaluated for immunogold-labeling of a trial natriuretic peptides (ANP) in rat right atria.Eight Wistar Kyoto retired breeders were sedated with pentobarbital, perfused with either 4% paraformaldehyde (LM) or Karnovsky's fixative (EM), and right atria were dissected, processed, embedded in paraffin or epon, respectively and sectioned according to conventional methods. For light microscopy, an indirect IGSS method according to Hacker (3) was performed. Paraffin sections on glass slides were washed in ddH2O, immersed in Lugol's iodine, washed in ddH2O and treated with 2.5% aqueous sodium thiosulfate for 20 sec. After additional washes in ddH2O and TBS-0.1% fish gelatin, 10% normal goat serum (PBS with 1% BSA) was applied for 20 min before an overnight incubation at 4°C with a polyclonal α-ANP primary antibody (Peninsula Labs, 1:1000 in TBS/BSA).


2011 ◽  
Vol 39 (10) ◽  
pp. 15
Author(s):  
MITCHEL L. ZOLER
Keyword(s):  

2015 ◽  
Vol 24 (2) ◽  
pp. 203-213 ◽  
Author(s):  
Federica Furfaro ◽  
Cristina Bezzio ◽  
Sandro Ardizzone ◽  
Alessandro Massari ◽  
Roberto De Franchis ◽  
...  

The treatment of ulcerative colitis (UC) has changed over the last decade. It is extremely important to optimize the therapies which are available nowadays and commonly used in daily clinical practice, as well as to stimulate the search for more powerful drugs for the induction and maintenance of sustained and durable remission, thus preventing further complications. Therefore, it is mandatory to identify the patients' prognostic variables associated with an aggressive clinical course and to test the most potent therapies accordingly.To date, the conventional therapeutic approach based on corticosteroids, salicylates (sulfasalazine, 5-aminosalicylic acid) or immunosuppressive agents is commonly used as a first step to induce and to maintain remission. However, in recent years, knowledge of new pathogenetic mechanisms of ulcerative colitis have allowed us to find new therapeutic targets leading to the development of new treatments that directly target proinflammatory mediators, such as TNF-alpha, cytokines, membrane migration agents, cellular therapies.The aim of this review is to provide the most significant data regarding the therapeutic role of drugs in UC and to give an overview of biological and experimental drugs that will become available in the near future. In particular, we will analyse the role of these drugs in the treatment of acute flare and maintenance of UC, as well as its importance in mucosal healing and in treating patients at a high risk of relapse.


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