Santa Cruz Biotechnology
anti-KLF4 antibody
sc-166238
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Based on literature mining, protein candidates are profiling to indicate if they are involved in mouse lung ischemia. KLF4 antibody from Santa Cruz is selected because of the price. As long as the antibody works, it would be good luck for the researcher to complete the project using the right antibody. To screen the proteins complicated in mouse lung ischemia, the lungs from B6 wild type mice were procured and subject to cold ischemia for 24 hours. The lung tissue lysates were made by adding lysis buffer [1% SDS, 10 mmol/L Tris-Cl (pH 7.6), 20 μg/μL aprotinin, 20 μg/μL leupeptin, and 1 mmol/L 4-(2-aminoethyl)benzenosulfonyl fluoride] after homogenizing with mortar and pestle in liquid nitrogen. The supernatants were obtained after centrifugation at 12000 g for 15 minutes at 4. The protein concentrations were determined using the Bicinchoninic Acid Protein Assay kit (Pierce, Rockford, IL). Fiften micrograms of protein were separated on SDS-PAGE gels and transferred to polyvinylidene difluoride membranes. After blocking, the membranes were incubated with the appropriately diluted primary antibody at 4°C overnight. After washing with TBST, the membranes were incubated with appropriately diluted HRP-conjugated secondary antibodies at room temperature for 1 hour. Proteins were detected with the enhanced chemiluminescence kit (Amersham Pharmacia Biotechnology, Inc., Piscataway, NJ).
Western Blot
Mouse lung
1/1000
5% milk
1 hour at room temperature
ECL
The expression levels of KLF4 was up-regulated after 24-hour cold ischemia
N/A
The antibody works
The system worked