Immunohistochemistry for CCL20 was performed as described by Verschuere et al [41 (link)]. Briefly, cryosections of terminal ileum were air-dried and fixed with ice-cold acetone. Endogenous peroxidase activity was quenched with 1% H2O2, followed by blocking of non-specific binding sites with 2% rabbit serum and 1% BSA in PBS. Subsequently, slides were incubated with the primary antibody (polyclonal goat anti-mouse CCL20, R&D Systems) or goat IgG isotype control (Santa Cruz Biotechnology, Santa Cruz, CA, USA) for 90 min at 37°C and with the biotinylated rabbit anti-goat secondary antibody (DAKO) for 30 min at room temperature. Then, HRP-conjugated streptavidin (DAKO) was applied for 30 min. DAB was used as enzyme substrate before counterstaining with haematoxilin.
Goat igg isotype control
Manufactured by Santa Cruz Biotechnology
Sourced in United States
Goat IgG isotype control is a laboratory reagent used as a control in immunological assays. It serves as a non-specific control to establish baseline levels of antibody binding or signal in the absence of a specific antigen.
Automatically generated - may contain errors
2 protocols using goat igg isotype control
1
Immunohistochemical Detection of CCL20 in Ileum
2
Immunofluorescence Staining of E-cadherin
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We performed immunofluorescence staining on 5-μm-thick cryosections that were prepared from formalin-fixed tissues and mounted on slides. For staining, we used anti-E-cadherin (R&D Systems) and goat IgG isotype control (Santa Cruz) as the primary antibodies and anti-goat Alexa 594 (Invitrogen) as the secondary antibody. Nuclei were counterstained with DAPI. Digital immunostaining images were captured and analyzed using an LSM780 confocal microscope (Carl Zeiss).
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