Immunohistochemistry and Immunocytochemistry

Antibody Solutions has generated many monoclonal and polyclonal antibodies for immunohistochemistry and immunocytochemistry applications as well as a number of optimized protocols. For example, we made a series of antibodies to human cyclooxygenase 2 (COX-2), one of which, AS66-P, is particularly useful for immunohistochemistry applications. COX-2 is a proinflammatory enzyme that is induced at sites of inflammation and dysregulated in many cancers. Our antibody was used for analysis of COX-2 expression in cystectomy biopsies of bladder cancer patients during a five-year follow up study. In this study, Liu et al (2001) utilized A66-P immunostaining of paraffin-embedded bladder tissue (left image) and showed that COX-2 expression in bladder tumor biopsies correlated with survival.
Using a monoclonal antibody custommade by Antibody Solutions against the Drosophila melanogaster homologue of p53, Ollmann et al. (Cell, 101:91, 2000) revealed an ancestral pro-apoptotic function for p53 and identified D. melanogaster as an ideal model system for elucidating the p53 apoptotic pathways induced by DNA damage.

Intracellular Cytokine Staining

Intracellular cytokine staining circumvents the need to isolate populations of cells by physical means such as panning methods, columns, density gradients, etc., which rarely yield populations of cells of sufficient purity that allow for easy data interpretation. The data presented above were generated by intracellular staining of peripheral blood mononuclear cells with our carboxyfluorescin-conjugated monoclonal antibody AS1-F to human tumor necrosis factor alpha (TNFα). This cytokine is secreted from many leukocytes in response to stimulation with bacterial lipopolysaccharide (LPS). In the left panel PBMC are gated into CD4+, CD16+ and other cells. When stimulated with LPS and Phorbol 12-Myristate-13-Acetate (PMA), only the CD16-positive cell population (mostly neutrophils) but not the CD4+ T-helper cell population responds with TNFα synthesis. Antibody Solutions offers a number of other cytokine-specific antibodies for flow cytometry applications, including AS5-F (human IL1-α), AS10-F (IL1-β), AS12-F (IL-6) and AS13-F (IL-8).

Intracellular Staining

Antibody Solutions has developed antibodies suitable for intracellular staining techniques. Our high-throughput flow cytometry screening platform is particularly advantageous for discovery of antibodies to intracellular proteins. Additionally, we are familiar with various fixation/permeabilization methods to use when evaluating antibody candidates. Antibody Solutions offers a number of cytokine-specific antibodies for intracellular flow cytometry applications, including AS5-F (human IL1-α), AS10-F (IL1-β), AS12-F (IL-6) and AS13-F (IL-8).