Total protein was isolated from the intestine by homogenization in cell lysis buffer (Sangon Biotech Co., Ltd., Shanghai, China), and then centrifuged at 12,000 × g for 30 min at 4 ℃. The proteins (5 μg/μL) were separated by 10% SDS-PAGE in Tris-glycine-SDS buffer. Separated proteins were transferred to NC membranes which were blocked with 5% skim milk (Bright Dairy Co., Ltd, China) for one hour at room temperature and then incubated with a 1:500 dilution of the individual primary antibodies anti-Slit2 (ab134166), anti-Robo4 (ab180824), anti-VEGF (ab150375), anti-NLRP3 (ab263899), anti-ASC (ab2236), anti-Caspase1 (ab207802), anti-ZO1 (ab61357), anti-Claudin5 (ab131259), anti-Occludin (ab167161), and anti-β-actin (ab6276) (Abcam, CA, USA). The antibodies were diluted in 3% BSA in Tris-buffered saline containing 0.1% Tween 20 (TBST), and applied overnight at 4℃. Membranes were washed and incubated with HRP-conjugated secondary antibodies (Proteintech, USA). Protein bands were detected using the Western Blot Luminol Reagent (Bio-rad, USA).
Anti slit2
Manufactured by Abcam
Sourced in China
Anti-Slit2 is a lab equipment product manufactured by Abcam. It is an antibody that detects the Slit2 protein. Slit2 is a secreted protein involved in axon guidance and cell migration.
Automatically generated - may contain errors
Lab products found in correlation
Hrp conjugated secondary antibody, by Proteintech (1 mentions)
Cell lysis buffer, by Sangon (1 mentions)
Anti β actin ab6276, by Abcam (1 mentions)
Anti robo4, by Abcam (1 mentions)
Chemiluminescent substrate, by Thermo Fisher Scientific (1 mentions)
Radioimmunoprecipitation assay buffer, by Thermo Fisher Scientific (1 mentions)
Anti n cadherin, by BD (1 mentions)
Anti e cadherin, by R&D Systems (1 mentions)
Anti vimentin, by Abcam (1 mentions)
Protease inhibitor cocktail, by Merck Group (1 mentions)
2 protocols using anti slit2
1
Intestinal Protein Profiling via Western Blot
2
Western Blot Analysis of EMT Markers
Check if the same lab product or an alternative is used in the 5 most similar protocols
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Whole-cell extracts were prepared with radioimmunoprecipitation assay buffer (Thermo Fisher Scientific) supplemented with protease inhibitor cocktail (Sigma-Aldrich). Proteins were separated on 4 to 15% precast protein gels (Bio-Rad) and transferred onto polyvinylidene difluoride membranes. Primary antibodies used were anti-SLIT2 (Abcam, ab134166), anti-Vimentin (Abcam, ab92547), anti–N-cadherin (BD Bioscience, 610920), anti–E-cadherin (R&D Systems, AF648), and anti-Snail (NEB, 3879S). Secondary antibodies were purchased from Cell Signaling Technology. Blots were visualized with chemiluminescent substrate (Thermo Fisher Scientific) using a ChemiDoc imaging system. Intensity of the protein bands were quantified using ImageJ.
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