AbCam
Anti-Biotin Rabbit Polyclonal antibody
ab53494
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We are performing experiments to develop new proteomics screening techniques using CRISPR. This antibody detects trace amounts of biotinylated proteins with great ease at the recommended dilution.
Western Blot
Cell Lysates from HEK293T and MEFs
1:10,000 Dilution in BSA, overnight incubation in cold room
1% BSA Room Temperature, 1 hour
Anti-Rabbit-HRP, 1:3000, 1 hour incubation at room temperature
NA
Biorad Clarity HRP substrate at 1:10 dilution
Protein extract from cells was prepared in RIPA lysis buffer supplied with PMSF, protease inhibitor mixture and sodium orthovanadate. Lysates were quantified using a BCA protein assay kit (Thermo). 20 micrograms of proteins were subjected to SDS-PAGE and transferred onto PVDF membranes and blocked with 1% BSA in 1xPBS + 0.2% TX100 at room temperature for 1 h with rocking. The membranes were probed with primary Antibody (1/10,000 dilution) in 1% BSA in 1xPBS + 0.2% TX100 overnight at 4°C. After washing with TBS-T, the membranes were incubated with HRP-conjugated secondary Antibodies in 5% non-fat milk/TBS-T at room temperature for 1 h. The protein–Antibody complex was detected by Biorad Clarity ECL substrate at 1:10 dilution in water.This antibody strongly detected biotinylated proteins in the cell lysates.
Must use BSA for blocking and primary antibody. Milk contains Biotin and will interfere with the antibody.
Extremely sensitive antobody
A little too strong
Great antibody for Biotin detection.