Quantifying Eotaxin Levels in BALF and Plasma

Eotaxin-1 and eotaxin-2 levels in BALF and plasma were quantified by using Quantikine ELISA Kits (R&D Systems Inc, Minneapolis, MN). Expression of eotaxins by proangiogenic hematopoietic progenitor cells was quantified using immunofluorescence as described in details elsewhere (30 (link)). Rabbit anti-mouse eotaxin-1 (1/50, Santa Cruz) and goat anti-mouse eotaxin-2 (1/50, Santa Cruz) polyclonal antibodies were used. Biotinylated donkey anti-rabbit (1/1,000 Jackson ImmunoResearch Laboratories, West Grove, PA) and streptavidin Alexa Fluor 568 (1/500, Life Technologies. Carlsbad, CA) were used as secondary and tertiary reagents, respectively, for eotaxin-1 detection. Donkey anti-goat Alexa Fluor 488 (1/500, Life Technologies) was used as secondary antibody for eotaxin-2. Values from different experiments were normalized against OVA/OVA group.

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Asosingh K., Vasanji A., Tipton A., Queisser K., Wanner N., Janocha A., Grandon D., Anand-Apte B., Rothenberg M.E., Dweik R, & Erzurum S.C. (2016). Eotaxin-rich Proangiogenic Hematopoietic Progenitor Cells and CCR3+ Endothelium in the Atopic Asthmatic Response. Journal of immunology (Baltimore, Md. : 1950), 196(5), 2377-2387.

Publication 2016

Quantikine ELISA Kits Biotinylated donkey anti-rabbit streptavidin Alexa Fluor 568 Donkey anti-goat Alexa Fluor 488

Alexa 568 Alexa fluor 488 Antibodies Antibody Donkey Elisa Eotaxin 1 Eotaxin 2 Goat Hematopoietic progenitor cells Immunofluorescence Mouse Plasma Rabbit Streptavidin

Corresponding Organization : Cleveland Clinic

Other organizations : Cleveland Eye Clinic, Cincinnati Children's Hospital Medical Center

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Variable analysis

independent variables

Eotaxin-1 and eotaxin-2 levels in BALF and plasma
Expression of eotaxins by proangiogenic hematopoietic progenitor cells

dependent variables

Eotaxin-1 and eotaxin-2 levels in BALF and plasma
Expression of eotaxins by proangiogenic hematopoietic progenitor cells

control variables

OVA/OVA group

controls

Positive controls: None specified
Negative controls: OVA/OVA group

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