Biotin labelled secondary antibodies

Manufactured by Agilent Technologies

Sourced in United Kingdom, Denmark

Biotin labelled secondary antibodies are designed for use in immunoassays and other research applications. They bind to the primary antibody and provide a means of detecting the target of interest through the biotin-streptavidin interaction.

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Lab products found in correlation

Streptavidin hrp complex, by Agilent Technologies (1 mentions) Phospho smad2 ser465 467, by Cell Signaling Technology (1 mentions) Superfrost plus microscope slide, by Thermo Fisher Scientific (1 mentions)

2 protocols using biotin labelled secondary antibodies

Detecting CYP450 Protein Expression

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Specific anti-peptide antibodies raised against individual CYP450 enzymes and the corresponding pre-immune sera have been previously described (Edwards et al, 1998 (link); Kapucuoglu et al, 2003 (link)). These antibodies have previously been used to detect human CYP450 protein expression (Piipari et al, 2000 (link), Rodriguez-Antona et al, 2002 (link)). Biotin labelled secondary antibodies and streptavidin/HRP complex were obtained from DAKO Ltd (Ely, UK). All other chemicals were supplied by either Sigma-Aldrich or Merck (Nottingham, UK) unless otherwise specified.

Patel S.A., Bhambra U., Charalambous M.P., David R.M., Edwards R.J., Lightfoot T., Boobis A.R, & Gooderham N.J. (2014). Interleukin-6 mediated upregulation of CYP1B1 and CYP2E1 in colorectal cancer involves DNA methylation, miR27b and STAT3. British Journal of Cancer, 111(12), 2287-2296.

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Immunohistochemical Analysis of Phospho-Smad2

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Samples were fixed overnight in phosphate buffered formalin, dehydrated and embedded in paraffin. Six mm thick sections were cut and mounted on Superfrost™ Plus Microscope Slides (Thermo Scientific, Waltham, USA). After deparaffinization, citrate buffer (0.1 M sodium citrate and 0.1 M citric acid) was used for 2 h at RT for antigen unmasking. Hydrogen peroxide 1% v/v in methanol was used for 30 min to block endogenous peroxidase. Afterwards, sections were incubated overnight at 4 C with specific primary antibodies against c-terminally phosphorylated SMAD2P (rabbit pAb anti Phospho-Smad2 (Ser465/467) (1:100) (Cell Signalling Technology, Danvers, Massachusetts, USA). Biotin-labelled secondary antibodies were used (Dako, Glostrup, Denmark). Together with a biotinestreptavidin detection system used according to the manufacturers' protocol (Vector Laboratories, Baiklin Game, California, USA). Staining was visualized using dimethylaminoazobenzene (DAB) reagent 23 (link) .

Madej W., van Caam A., Blaney Davidson E., Buma P, & van der Kraan P.M. (2016). Unloading results in rapid loss of TGFβ signaling in articular cartilage: role of loading-induced TGFβ signaling in maintenance of articular chondrocyte phenotype?. Osteoarthritis and cartilage, 24(10).

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