Images
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Western blot analysis of Myc tag on different lysates with Mouse anti-Myc tag antibody (HA601513) at 1/20,000 dilution.
Lane 1: 293T transfected with Myc-tagged ACAT2 (C-terminal) cell lysate
Lane 2: 293T transfected with Myc-tagged Claudin18.2 (C-terminal) cell lysate
Lane 3: 293T transfected with Myc-tagged Histone H3.1 (N-terminal) cell lysate
Lysates/proteins at 10 µg/Lane.
Exposure time: 25 seconds; ECL: K1801;
4-20% SDS-PAGE gel.
Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (HA601513) at 1/20,000 dilution was used in primary antibody dilution (K1803) at 4℃ overnight. Goat Anti-Mouse IgG - HRP Secondary Antibody (HA1006) at 1/50,000 dilution was used for 1 hour at room temperature.
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Immunocytochemistry analysis of HeLa cells labeling Myc tag with Mouse anti-Myc tag antibody (HA601513) at 1/200 dilution.
HeLa cells, transfected with empty control (top, negative) / Myc-tagged Claudin18.2 (C-terminal) (middle, positive) / Myc-tagged Histone H3.1 (N-terminal) (bottom, positive) expression vector, respectively, were fixed in 100% precooled methanol for 5 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Mouse anti-Myc tag antibody (HA601513) at 1/200 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Mouse IgG H&L (iFluor™ 488, HA1125) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI.
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Myc tag was immunoprecipitated from 0.2 mg 293T transfected with Myc-tagged Histone H3.1 (N-terminal) cell lysate with HA601513 at 2 µg/10 µl beads. Western blot was performed from the immunoprecipitate using HA601513 at 1/20,000 dilution. Anti-Mouse IgG for IP Nano-secondary antibody (NBI02H) at 1/5,000 dilution was used for 1 hour at room temperature.
Lane 1: 293T transfected with Myc-tagged Histone H3.1 (N-terminal) cell lysate (input)
Lane 2: HA601513 IP in 293T transfected with Myc-tagged Histone H3.1 (N-terminal) cell lysate
Lane 3: Mouse IgG instead of HA601513 in 293T transfected with Myc-tagged Histone H3.1 (N-terminal) cell lysate
Blocking/Dilution buffer: 5% NFDM/TBST
Exposure time: 10 seconds; ECL: K1801
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