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Western blot analysis of Glucosidase 2 subunit beta on different lysates with Mouse anti-Glucosidase 2 subunit beta antibody (HA600054) at 1/1,000 dilution.
Lane 1: HeLa cell lysate
Lane 2: 293T cell lysate
Lane 3: Daudi cell lysate
Lane 4: Jurkat cell lysate
Lane 5: A431 cell lysate
Lysates/proteins at 20 µg/Lane.
Predicted band size: 59 kDa
Observed band size: 80 kDa
Exposure time: 5 minutes;
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 (HA600054) at 1/1,000 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 Hela cells labeling Glucosidase 2 subunit beta with Mouse anti-Glucosidase 2 subunit beta antibody (HA600054) at 1/100 dilution.
Cells were fixed in 4% paraformaldehyde for 15 minutes, permeabilized with 0.1% Triton X-100 in PBS for 15 minutes, and then blocked with 1% BSA for 30 minutes at room temperature. Cells were then incubated with Mouse anti-Glucosidase 2 subunit beta antibody (HA600054) at 1/100 dilution in 1% BSA 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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Immunohistochemical analysis of paraffin-embedded human placenta tissue with Mouse anti-Glucosidase 2 subunit beta antibody (HA600054) at 1/200 dilution.
The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HA600054) at 1/200 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
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Flow cytometric analysis of Hela cells labeling Glucosidase 2 subunit beta.
Cells were fixed and permeabilized. Then stained with the primary antibody (HA600054, 1ug/ml) (red) compared with Mouse IgG1 Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a iFluor™ 488 conjugate-Goat anti-Mouse IgG Secondary antibody (HA1125) at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).
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