Immunofluorescence staining was performed [43 (link)] to evaluate ZO-1 and occludin expression in the colon. Anti-TSLP (rabbit, 1:50; GTX85059, GeneTex, Irvine, CA, USA), anti-IL-6R (rabbit, 1:1000; Invitrogen, Waltham, MA, USA), anti-ZO-1 (rabbit, 1:500; 61–7300, Invitrogen, Waltham, MA, USA), and anti-occludin (mouse, 1:200; OC3F10, Invitrogen, Waltham, MA, USA) antibodies were diluted in 1× PBST supplemented with 0.3% BSA. The slides were wrapped in an aluminum foil to block light and stored at 4 °C for 72 h. Tissue sections were incubated for 1 h with a mixture of Alexa 488-conjugated donkey anti-mouse secondary antibody (1:1000; A21206, Invitrogen, Waltham, MA, USA) or Alexa 594-conjugated donkey anti-mouse secondary antibody (1:1000; A21203, Invitrogen, Waltham, MA, USA).
Gtx85059
Manufactured by GeneTex
Sourced in United States
The GTX85059 is a multi-channel pipette designed for accurate and efficient liquid handling. It features adjustable volumes and ergonomic controls for improved workflow in laboratory settings.
Automatically generated - may contain errors
Lab products found in correlation
A21206, by Thermo Fisher Scientific (1 mentions)
A21203, by Thermo Fisher Scientific (1 mentions)
Oc 3f10, by Thermo Fisher Scientific (1 mentions)
Alexa fluor dye, by Thermo Fisher Scientific (1 mentions)
Prolong gold anti fade kit, by Thermo Fisher Scientific (1 mentions)
Dihydroethidium dhe, by Thermo Fisher Scientific (1 mentions)
Ds fi2 camera, by Nikon (1 mentions)
Eclipse ti u microscope, by Nikon (1 mentions)
2 protocols using gtx85059
1
Immunofluorescence Staining of Tight Junctions
2
Immunofluorescence Staining of Lung Tissues
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For immunofluorescence staining, sectioned lung tissues were first blocked using 5% w/v BSA for 1 hour, followed by incubation with antibodies against TSLP (GTX85059; GeneTex), IL‐33 (AF3626; R&D Systems), IL‐25 (207710; R&D Systems), and EpCAM (G8.8; ThermoFisher) overnight at 4℃. Sample sections were then incubated with secondary antibodies conjugated with Alexa Fluor dyes (ThermoFisher) at room temperature for 1 hour. Isotype‐matched negative control antibodies (R&D Systems) were used under the same conditions. Nuclei were counterstained with 6‐diamidino‐2‐phenylindole, dihydrochloride (DAPI, ThermoFisher). Sections were mounted with the ProLong Gold Anti‐fade Kit (Molecular Probes) and observed with a Nikon Eclipse Ti‐U microscope equipped with a DS‐Fi2 camera (Nikon). The detection of intracellular superoxide in lung tissues was carried out using dihydroethidium (DHE) (ThermoFisher). To determine the fluorescence signal in tissue sections, fluorescent‐positive cells in five different high‐power fields from each slide were quantified using ImageJ v1.50e (NIH) and presented as mean fluorescence intensity per square micrometer. Two to three slides from each sample were used for analysis.
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