Streptavidin biotin detection system

Manufactured by Biocare Medical

Sourced in United States

The Streptavidin biotin detection system is a laboratory instrument used to detect and analyze the presence and quantity of specific biomolecules in a sample. It utilizes the high-affinity interaction between streptavidin and biotin to facilitate the identification and quantification of target analytes.

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Lab products found in correlation

S1699, by Agilent Technologies (1 mentions) Mib 1, by Agilent Technologies (1 mentions) Cd3 12, by Bio-Rad (1 mentions)

3 protocols using streptavidin biotin detection system

Feline Infectious Peritonitis Histopathology

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Samples of tissues obtained at necropsy were immediately placed in 10% buffered formalin and fixed for 24 h before being routinely embedded into paraffin blocks. Tissues that were examined histologically (all cats) and by immunohistochemistry (two cats) included liver, spleen, kidney, mesenteric lymph node, omentum, ileum, caecum, heart, and lungs.
Immunohistochemistry was performed on 4 μm serial sections of formalin fixed and paraffin embedded tissues using a streptavidin biotin detection system (Biocare Medical, Concord, CA). Mouse monoclonal antibodies to FIPV (Custom Monoclonal International, clone FIPV3-70, 1:200), CD18 (Peter Moore, UC Davis, clone Fe3.9F2), and CD3 (Peter Moore, UC Davis, clone CD3-12) were used with some modifications. Before applying the primary antibodies, slides were steam pretreated in citrate buffer (Dako S1699) at 98 °C for 20 min followed by a 20 min cooling. They were then washed in PBS and blocked with 10% normal horse serum for 20 min. Amino ethyl carbazole (AEC, Dako Corp.) was used as the chromogen sections were counterstained with Mayer's hematoxylin. Substituting a matched mouse IgG correlate for the primary antibody served as the negative control.

Pedersen N.C., Eckstrand C., Liu H., Leutenegger C, & Murphy B. (2014). Levels of feline infectious peritonitis virus in blood, effusions, and various tissues and the role of lymphopenia in disease outcome following experimental infection. Veterinary Microbiology, 175(2), 157-166.

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In-Situ Hybridization for Viral RNA Detection

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An in-situ hybridization technique was used to visualize the presence, location, and quantity of tissue associated viral nucleic acid in the following tissues: cervical LNs 1 and 2 (lymphoma), mesenteric LN (lymphoid atrophy), small intestine, and bone marrow. The assay was performed on 5 μm sections of formalin-fixed paraffin-embedded tissue set on positively charged glass slides, according to manufacturer protocols, using the RNAscope 2.5 Brown kit (Advanced Cell Diagnostics, Newark, CA, USA) and a nucleic acid probe designed to hybridize FIV-C-specific RNA (cat# 462951) [5 (link)]. Spleen, lymph node, and gut tissues from an age-matched sham-infected SPF cat were used as a negative control [5 (link)]. A probe specific for a feline housekeeping gene RNA (Felis catus peptidylprolyl isomerase B) and for the Bacillus subtilis strain SMY dihydrodipicolinate reductase (dapB, cat #310043) gene were used as positive and negative controls, respectively.
Immunohistochemistry (IHC) assays detecting CD3 antigen (clone CD3-12, P. Moore, Davis, CA, USA) were performed on 5 μm-thick, FFPE tissue sections (cervical LN 1 and 2, mesenteric lymph node, small intestine, and bone marrow) using a streptavidin biotin detection system (Biocare Medical, Pacheco, CA, USA), as described previously [26 (link)].

Murphy B.G., Eckstrand C., Castillo D., Poon A., Liepnieks M., Harmon K, & Moore P. (2018). Multiple, Independent T Cell Lymphomas Arising in an Experimentally FIV-Infected Cat during the Terminal Stage of Infection. Viruses, 10(6), 280.

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Immunophenotyping of Lymph Node Leukocytes

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Immunohistochemistry (IHC) was utilized to phenotype PLN leukocytes using antibodies for feline CD3 (Peter Moore, UC Davis, clone CD3-12), CD79 (AbD Serotec, clone HM57), CD204 (Trans Genic, clone SRA-E5), and Ki67 (Dako, clone MIB-1) on 4 micron serial sections with a streptavidin biotin detection system (Biocare Medical) and following the protocol described above, but using an anti-mouse secondary antibody. Substituting a matched mouse IgG for the primary antibody served as the negative control. The proportion of CD3+ T cells within each lymphoid follicle and Ki67+ cells in the paracortical zones were quantified utilizing ImageJ software (National Institutes of Health). Briefly, photomicrographs of lymphoid follicles and paracortical zones (five different follicles/cat and three random paracortical regions/cat) were obtained. Using ImageJ, the image threshold was adjusted to black (IHC positive cells) and white to minimize non-specific background. IHC positive cells were quantified by specifying the lower limit of cell size (100 pixels) and accounting for overlapping cells. The image was then assessed for total number of nuclei using the image-based tool for counting nuclei (ITCN). The frequency of IHC positive cells was expressed as a division of the number of positive cells divided by total number of nuclei.

Eckstrand C.D., Hillman C., Smith A.L., Sparger E.E, & Murphy B.G. (2016). Viral Reservoirs in Lymph Nodes of FIV-Infected Progressor and Long-Term Non-Progressor Cats during the Asymptomatic Phase. PLoS ONE, 11(1), e0146285.

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