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Anti-HA.11 Epitope Tag Antibody (Mouse)
901514
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We are performing experiments to develop new proteomics screening techniques using CRISPR. This antibody detects HA epitope tagged proteins with great ease in a variety of human and mouse cell types.
Western Blot
Cell Lysates from HEK293T and MEFs
1:1000 Dilution in BSA, overnight incubation in cold room
5% non-fat milk in TBS-T, 1hour at Room Temp
Anti-Mouse-HRP, 1:3000, 1hour incubation at Room Temperature
NA
Bio-Rad Clarity HRP substrate
Protein extract from cells was prepared in RIPA lysis buffer supplied with PMSF, protease inhibitor mixture and sodium orthovanadate. Lysates were quantified using BSA assay kit (Thermo). 20 micrograms of proteins were subjected to SDS-PAGE and transferred onto PVDF membranes and blocked with 5% non-fat milk in TBS-T (20 mMTris, 500 mMNaCl, and 0.1% Tween 20) at room temperature for 1 h with rocking. The membranes were probed with primary Antibody (1/1000 dilution) 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.This antibody detected HA-tagged dCas9 protein in HEK293T cells and Mouse Embryonic Fibroblast Cells.
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Excellent antibody for detecting HA epitope tag
This antibody easily detects HA epitope tag in a variety of cell line and in yeast. It can be reused several times if the diluted antibody is frozen down after each use.