Stomachs were prepared, stained and imaged using methods modified from Ramsey et al.42 (link) Briefly, paraffin-embedded specimens were cut into 5 μm sections, deparaffinised and rehydrated. After antigen retrieval with 10 mM sodium citrate (pH 6.0), sections were washed with PBS and blocked in 1% BSA and 0.3% Triton X-100 in PBS followed by overnight incubation with primary antibodies. The primary antibodies used for immunostaining were rat anti-CD44v (1:10 000 from Cosmo Bio, LKGM002) and goat anti-GIF (1:10,000, gift from Dr David Alpers, Washington University in St. Louis). After washing, sections were incubated with secondary antibodies and GS-II lectin (1:500, L21415, ThermoFisher). Sections were washed, stained with Hoechst (62249, ThermoFisher) 1:20 000 in PBS and mounted in ProLong Gold Antifade mountant (P36934, ThermoFisher).
Lkgm002
Manufactured by Cosmo Bio
The LKGM002 is a laboratory centrifuge designed for general-purpose applications. It features a compact and durable construction, providing consistent and reliable performance. The centrifuge can accommodate a range of sample volumes and tube sizes, making it a versatile tool for various laboratory procedures.
Automatically generated - may contain errors
Lab products found in correlation
Hoechst, by Thermo Fisher Scientific (3 mentions)
L21415, by Thermo Fisher Scientific (3 mentions)
Prolong gold antifade mountant, by Thermo Fisher Scientific (3 mentions)
Sc 13083, by Santa Cruz Biotechnology (2 mentions)
Bovine serum albumin (bsa), by Avantor (1 mentions)
Cytospin 4 centrifuge, by Thermo Fisher Scientific (1 mentions)
Bs 2606r, by Bioss Antibodies (1 mentions)
4 protocols using lkgm002
1
Immunostaining of Stomach Tissues
2
Immunostaining of Gastric Tissues
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Stomachs were prepared, stained, and imaged using methods modified from Ramsey et al.57 (link) The primary antibodies used for immunostaining were goat anti–vascular endothelial growth factor B (1:100, sc-13083; Santa Cruz Biotechnology, Dallas, TX), anti-CD44v9 (1:10,000, LKGM002; Cosmo Bio, Carlsbad, CA), and goat anti-GIF (1:10,000; a gift from David Alpers, Washington University in St. Louis). After washing, sections were incubated with secondary antibodies and GS-II lectin (1:500, L21415; ThermoFisher, Waltham, MA). Sections were washed, stained with Hoechst (62249; ThermoFisher) 1:20,000 in PBS, and mounted in ProLong Gold Antifade mountant (P36934; ThermoFisher).
3
Stomach Tissue Immunostaining Protocol
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Stomachs were prepared, stained and imaged using methods modified from Ramsey et al.42 (link) Briefly, paraffin-embedded specimens were cut into 5 µm sections, deparaffinised and rehydrated. After antigen retrieval with 10 mM sodium citrate (pH 6.0), sections were washed with PBS and blocked in 1% BSA and 0.3% Triton X-100 in PBS followed by overnight incubation with primary antibodies. The primary antibodies used for immunostaining were rat anti-CD44v (1:10 000 from Cosmo Bio, LKGM002) and goat anti-GIF (1:10,000, gift from Dr David Alpers, Washington University in St. Louis). After washing, sections were incubated with secondary antibodies and GS-II lectin (1:500, L21415, ThermoFisher). Sections were washed, stained with Hoechst (62249, ThermoFisher) 1:20 000 in PBS and mounted in ProLong Gold Antifade mountant (P36934, ThermoFisher).
4
Immunohistochemical Analysis of Stomach Tissues
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Stomachs were prepared, stained, and imaged using methods modified from Ramsey et al.35 (link) The primary antibodies used for immunostaining were as follows: rabbit anti-IL-17RA (1:100, bs-2606R; BIOSS, Woburn, MA), goat anti–vascular endothelial growth factor B (VEGF-B) (1:100, sc-13083; Santa Cruz Biotechnology, Dallas, TX), and anti-CD44v9 (1:10,000, LKGM002; Cosmo Bio, Carlsbad, CA). Secondary antibody labeling was as described. For activated caspase-3 immunohistochemistry, a Cell Signaling Technologies (Danvers, MA) SignalStain Apoptosis Kit (12692S) was used according to the manufacturer’s protocol. For terminal deoxynucleotidyl transferase–mediated deoxyuridine triphosphate nick-end labeling (TUNEL) staining, an In Situ Death Detection Kit (11684795910; Millipore-Sigma, St. Louis, MO) was used according to the manufacturer’s specifications. Slides of glands for immunofluorescence were generated using a Cytospin 4 centrifuge (A78300003; ThermoFisher, Waltham, MA). Cells were fixed on the slide in 4% paraformaldehyde for 20 minutes at room temperature and permeabilized (0.5% bovine serum albumin, 0.1% Triton (VWR International, Radnor, PA), and 2 mmol/L EDTA in phosphate-buffered saline) for 30 minutes at room temperature before blocking and staining according to the earlier-described protocols.
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