Antimicrobial peptide PR39 was synthesized by GL Biochem (Shanghai China), and the purity was more than 95% by reversed-phase high performance liquid chromatography analysis. the peptide was prepared in sterile saline and stored under −80 °C until used. Other normal reagents were listed on the paper we have previously published51 (link), to introduce briefly: Rabbit polyclonal Abs for zonula occludens (ZO-1), Occludin, CD177 and F4/80 were purchased from Abcam (Cambridge, MA). The secondary Abs used for the immunohistochemistry was goat anti-rabbit IgG conjugated with HRP (Abcam) and for the immunofluorescence were goat anti-rabbit IgG labeling with FITC and Cy3 (Abcam), the TUNEL kit was purchased from Roche (USA). The EHEC O157:H7 ATCC43889 was purchased from China General Microbiological Culture Collection Center (Beijing, China).
Cd177
Manufactured by Abcam
Sourced in United Kingdom
CD177 is a surface glycoprotein expressed on the surface of neutrophils. It functions as a receptor for the urokinase-type plasminogen activator and may play a role in neutrophil activation and migration.
Automatically generated - may contain errors
Lab products found in correlation
Tunel kit, by Roche (1 mentions)
Goat anti rabbit igg conjugated with hrp, by Abcam (1 mentions)
F4 80, by Abcam (1 mentions)
Horseradish peroxidase hrp conjugated goat anti rabbit secondary antibody, by Abcam (1 mentions)
Pierce detergent compatible bradford assay kit, by Thermo Fisher Scientific (1 mentions)
3 protocols using cd177
1
Antimicrobial Peptide PR39 Protocol
2
Immunohistochemical Analysis of Uveitis in Mice
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The mice were sacrificed by cervical dislocation at 12, 24, 48 and 72 h after LPS-induced uveitis. The enucleated eyes were fixed with 4% paraformaldehyde, embedded in paraffin, cut into 5-µm serial sections and deparaffinized. Antigen retrieval was performed by heating sections in a microwave oven in citrate buffer [0.01 mol/l (pH 6.0)] for 30 min. Endogenous peroxidase was eliminated by incubation in 3% hydrogen peroxide in PBS for 10 min, and the tissue sections were washed in PBS for 5 min three times, and incubated in 10% goat serum (Biosharp, Hefei, China) for 2 h. The tissue sections were incubated with rabbit polyclonal antibodies against cluster of differentiation 177 (CD177; dilution, 1:100; Abcam, Cambridge, UK; cat. no. ab203025) overnight at 4°C, followed by a goat anti-rabbit horseradish peroxidase (HRP)-conjugated secondary antibody (dilution, 1:1,000; Abcam; cat. no. ab97200) for 1 h at 37°C and stained with peroxidase. The stained cells in the vitreous body near the optic nerve head were observed under light microscopy (n=6 per group).
3
Protein Quantification and Western Blot
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The tissues were lysed, and total protein was quantified using the PierceTM Detergent Compatible Bradford Assay Kit (Thermo Scientific). 20 μg of protein from each sample was used for SDS-PAGE. After transferring the sample onto a PVDF membrane, the blot was incubated with indicate antibodies. All antibodies were purchased from CST: CD177 (ab203025, Abcam), TTPAL (ab103740, Abcam), FLT3 (#3462, CST), and GAPDH (#5174, CST).
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