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☑ Cell treatment (CT)
Western blot analysis of mCherry on different lysates with Mouse anti-mCherry antibody (HA601186) at 1/2,000 dilution.
Lane 1: 293T-WT cell lysate (5 µg/Lane)
Lane 2: 293T-OE-mCherry cell lysate (5 µg/Lane)
Lane 3: 293T-OE-mCherry cell lysate (2.5 µg/Lane)
Lane 4: 293T-OE-mCherry cell lysate (1.25 µg/Lane)
Lane 5: 293T-OE-mCherry cell lysate (0.625 µg/Lane)
Predicted band size: 27 kDa
Observed band size: 27 kDa
Exposure time: 30 seconds;
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 (HA601186) at 1/2,000 dilution was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Mouse IgG - HRP Secondary Antibody (HA1006) at 1/150,000 dilution was used for 1 hour at room temperature.
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Western blot analysis of mCherry on mCherry recombinant protein with Mouse anti-mCherry antibody (HA601186) at 1/1,000 dilution.
Lane 1: mCherry recombinant protein, 500ng/Lane
Lane 2: mCherry recombinant protein, 250ng/Lane
Lane 3: mCherry recombinant protein, 125ng/Lane
Lane 4: mCherry recombinant protein, 62.5ng/Lane
Exposure time: 30 seconds;
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 (HA601186) at 1/1,000 dilution was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Mouse IgG - HRP Secondary Antibody (HA1006) at 1/150,000 dilution was used for 1 hour at room temperature.
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Western blot analysis of mCherry on different lysates with Mouse anti-mCherry antibody (HA601186) at 1/5,000 dilution.
Lane 1: 293T cell lysate
Lane 2: 293T transfected with mCherry-tagged cell lysate
Lane 3: 293T transfected with GFP-tagged cell lysate (negative)
Lysates/proteins at 5 µg/Lane.
Exposure time: 3 minutes; 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 (HA601186) at 1/5,000 dilution was used in 5% NFDM/TBST 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 293T transfected with or without mCherry cells labeling mCherry with Mouse anti-mCherry antibody (HA601186) at 1/2,000 dilution.
Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS 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-mCherry antibody (HA601186) at 1/2,000 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. As expected, green antibody staining is only visible in cells expressing mCherry, and green and red signals can be superimposed.
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