Intestinal specimens were harvested and fixed in 4% paraformaldehyde prior to cryoprotection, OCT embedding and cryosectioning. Sections (10 μm) were stained using various IEC subtype-specific antibodies or lectins: rat-anti-E-cadherin (all IECs; ECCD-2, Takara, #M108, Mountain View, CA, USA), goat-anti-Anterior Gradient 2 (goblet cells; Santa Cruz, #sc54561, Dallas, Texas, USA), rabbit-anti-Chromogranin A (enteroendocrine cells; ImmunoStar, #20086, Hudson, WI, USA), Ulex europaeus lectin-FITC (Paneth cells; Sigma-Aldrich, #L9006, St. Louis, MO, USA). Secondary antibodies were donkey-anti-rat-AF594 (Life Technologies, #A21209 Carlsbad, CA, USA), donkey-anti-goat-Cy3 (Jackson Immuno Research, #705-166-147, West Grove, PA, USA) and donkey-anti-rabbit-BV421 (Nordic BioSite, #406410, Täby, Stockholm, Sweden). HOECHST (Life Technologies, #H1399) was used as DNA counterstaining. Slides were scanned using Metafer automated slide scanner (MetaSystems, Altlussheim, Baden-Württemberg, Germany) and composite pictures analyzed using the Visiopharm Integrator System program [16 (link)]. Total area for each staining was determined for complete intestinal sections and expressed as percentage of epithelial area (E-cadherin).
Donkey anti rat af594
Manufactured by Thermo Fisher Scientific
Sourced in United States
Donkey-anti-rat-AF594 is a secondary antibody conjugated with the fluorescent dye Alexa Fluor 594. It is designed to detect and visualize primary antibodies raised against rat antigens in various immunoassays and imaging applications.
Automatically generated - may contain errors
Lab products found in correlation
Eccd 2, by Takara Bio (1 mentions)
Hoechst, by Thermo Fisher Scientific (1 mentions)
Donkey anti goat cy3, by Jackson ImmunoResearch (1 mentions)
Rat anti e cadherin, by Takara Bio (1 mentions)
Ab107099, by Abcam (1 mentions)
Af594 donkey anti rabbit, by Thermo Fisher Scientific (1 mentions)
Donkey anti goat af488, by Thermo Fisher Scientific (1 mentions)
Click it edu alexa fluor 594 imaging kit, by Thermo Fisher Scientific (1 mentions)
Mec13, by BD (1 mentions)
2 protocols using donkey anti rat af594
1
Intestinal Epithelial Cell Subtype Profiling
2
Multimodal Immunohistochemistry Analysis
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Immunohistochemistry was performed as described previously (Richardson et al., 2012 ). 10 μm sections were used for all studies. Primary antibodies used: rat ant‐p21 (HUGO291, Abcam ab107099), goat‐anti‐troponin C (Abcam, ab30807), rabbit anti‐p16 (Rockland, 100‐401‐170) and rat anti‐CD31 (MEC13.3, BD Biosciences, 550274). Secondary antibodies used were donkey anti‐rat AF594 (Life Technologies, A21209), donkey anti‐goat AF 488 nm (Life Technologies, A11055), donkey anti‐rabbit AF594 (Life Technologies, R37119), donkey anti‐goat AF488 (Life Technologies, A11055) and donkey anti‐mouse AF647 (A31571, Life Technologies). Slides were mounted in Vectarshield containing DAPI (Sigma, MBD0015). 5‐ethynyl‐2‐deoxyuridine (EdU) labeling performed with Invitrogen Click‐iT EdU Alexa Fluor 594 Imaging Kit (Life Technologies, C10339).
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