ST2 Mouse Monoclonal Antibody [PSH01-68]

☑ Cell treatment (CT)

Western blot analysis of ST2 on different lysates with Mouse anti-ST2 antibody (HA601170) at 1/1,000 dilution.

Lane 1: HeLa transfected with FLAG-tagged empty control cell lysate
Lane 2: HeLa transfected with FLAG-tagged ST2 cell lysate

Lysates/proteins at 30 µg/Lane.

Predicted band size: 63 kDa
Observed band size: 80 kDa

Exposure time: 5 minutes 10 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 (HA601170) 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 1/50,000 dilution was used for 1 hour at room temperature.

☑ Cell treatment (CT)

Immunocytochemistry analysis of HeLa overexpress with or without ST2 cells labeling ST2 with Mouse anti-ST2 antibody (HA601170) at 1/500 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-ST2 antibody (HA601170) at 1/500 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.

beta Tubulin (ET1602-4, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Rabbit IgG H&L (iFluor™ 594, HA1122) were used as the secondary antibody at 1/1,000 dilution.

☑ Cell treatment (CT)

Flow cytometric analysis of HeLa overexpress with or without ST2 cells labeling ST2.

Cells were washed twice with cold PBS and resuspend. Then stained with the primary antibody (HA601170, 1/1,000) (red) compared with Mouse IgG1 Isotype Control (green). After incubation of the primary antibody at +4℃ for 30 minutes, the cells were stained with a iFluor™ 594 conjugate-Goat anti-Mouse IgG Secondary antibody (HA1126) at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).