The protein lysate was obtained from culture cells by culture cell total protein extraction reagent (BOSTER, Wuhan, CHN) with protease inhibitor and phosphatase inhibitor cocktail. Protein concentration was determined by a BCA protein analysis kit (BIOMED, Beijing, CHN). Primary antibodies included rabbit anti-β-actin (#20536-1-AP; Proteintech, Wuhan, CHN), rabbit anti-ZO-1 (#21773-1-AP; Proteintech, Wuhan, CHN), rabbit anti-Occludin (#27260–1-AP; Proteintech, Wuhan, CHN), rabbit anti-Claudin 1 (#bs-1428R; Bioss, Beijing, CHN), rabbit anti-NF-κB P65 (#bs-0465R; Bioss, Beijing, CHN) and rabbit anti-phospho-NF-κB p65 (#3033 T; CST, Danvers, MA USA). Goat anti-rabbit (#bs-0296G; Bioss, Beijing, CHN) was used as secondary antibody.
Bs 1428r
Manufactured by Bioss Antibodies
Sourced in United States
The Bs-1428R is a laboratory instrument designed for the detection and analysis of biomolecules. It utilizes a sensitive detection method to provide reliable and accurate results. The core function of the Bs-1428R is to facilitate the quantitative and qualitative evaluation of various biological samples.
Automatically generated - may contain errors
Lab products found in correlation
Culture cell total protein extraction reagent, by Boster Bio (1 mentions)
Bs 0296g, by Bioss Antibodies (1 mentions)
Rabbit anti zo 1, by Proteintech (1 mentions)
Rabbit anti occludin, by Proteintech (1 mentions)
Rabbit anti nf κb p65, by Bioss Antibodies (1 mentions)
Rabbit anti claudin 1, by Bioss Antibodies (1 mentions)
Rabbit anti β actin, by Proteintech (1 mentions)
Occludin, by Bioss Antibodies (1 mentions)
F actin, by Bioss Antibodies (1 mentions)
E cadherin, by Bioss Antibodies (1 mentions)
Ecl chemiluminescence substrate, by Biosharp (1 mentions)
High efficiency ripa lysate, by Solarbio (1 mentions)
Las4000, by GE Healthcare (1 mentions)
Sa00001 2, by Proteintech (1 mentions)
Total protein assay kit, by Nanjing Jiancheng (1 mentions)
3 protocols using bs 1428r
1
Protein Extraction and Western Blot
2
Ileum Tight Junction Protein Analysis
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After deparaffinization, the ileum sections were incubated in citrate buffer (pH: 6.0) (Lab Vision, Thermo Scientific, Fremont) and 3% hydrogen peroxide (Lab Vision, Thermo Scientific, Fremont). After washing with PBS (Thermo, AP-9009-10, UK) (pH: 7.4), Ultra V block (Lab Vision, Thermo Scientific, Fremont) was applied to prevent non-specific binding. Following the blocking stage, tissue sections were incubated with Zonulin (bs-1808R, Bioss), Occludin (bs-1495R, Bioss), Claudin-1 (bs-1428R, Bioss), Claudin-2 (bs-7125R, Bioss), Zonula Occludens-1 (ZO-1) (bs-1329R, Bioss), E-Cadherin (bs-10009R, Bioss), and F-Actin (bs-1571R, Bioss) primer antibodies (1:200 dilution) overnight. Then, the sections were incubated with secondary antibodies (Lab Vision, Thermo Scientific, Fremont) for 10 min. The reaction product was revealed by streptavidin peroxidase complex (Lab Vision, Thermo Scientific, Fremont) with Diaminobenzidine (DAB). The percentage of immunopositivity of primer antibodies was determined in each area using the ImageJ (Java-based software program, National Institutes of Health).
3
Quantifying Tight Junction Proteins in Mice
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The samples were prepared by grinding the colonic tissue of experimental mice with high-efficiency RIPA lysate (Solarbio, Beijing, China) supplemented with protease inhibitors. A total protein assay kit (Jiancheng, Nanjing, China) was used to determine the protein content of the samples. The samples were separated by polyacrylamide gel electrophoresis and transferred to a polyvinylidene difluoride membrane. The membrane was sealed with 5% skim milk powder for 1 h at room temperature. After incubation, proteins on the membrane were detected with ECL chemiluminescence substrate (Biosharp, HeFei, China) and blotted on a gel imaging system (LAS4000, GE, Boston, MA, USA). Primary antibodies against Claudin-1 (Bioss, Woburn, MA, USA, bs-1428R, 1/1000), Occludin (Bioss, bs-10011R, 1/1000), SOCS1 (Abcepta, San Diego, CA, USA, AP8790A, 1/1000), β-actin (Proteintech, Rosemont, IL, USA 20536-1-AP, 1/2000) and secondary antibody (Proteintech, SA00001-2, 1/5000) were used for Western blot analysis. The intensity of the protein bands was analyzed using image analysis software (Image J 1.53a, Bethesda, MD, USA).
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