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Western blot analysis of beta Tubulin on different lysates with Mouse anti-beta Tubulin antibody (HA601187) at 1/10,000 dilution.
Lane 1: HepG2 cell lysate (10 µg/Lane)
Lane 2: A549 cell lysate (10 µg/Lane)
Lane 3: NIH/3T3 cell lysate (10 µg/Lane)
Lane 4: L-929 cell lysate (10 µg/Lane)
Lane 5: PC-12 cell lysate (10 µg/Lane)
Lane 6: C6 cell lysate (10 µg/Lane)
Lane 7: Vero cell lysate (10 µg/Lane)
Lane 8: COS-1 cell lysate (10 µg/Lane)
Lane 9: Mouse spleen tissue lysate (20 µg/Lane)
Lane 10: Mouse stomach tissue lysate (20 µg/Lane)
Lane 11: Rat kidney tissue lysate (20 µg/Lane)
Predicted band size: 50 kDa
Observed band size: 50 kDa
Exposure time: 24 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 (HA601187) at 1/10,000 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.
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Immunocytochemistry analysis of HeLa cells labeling beta Tubulin with Mouse anti-beta Tubulin antibody (HA601187) at 1/100 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-beta Tubulin antibody (HA601187) at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) 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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Flow cytometric analysis of HeLa cells labeling beta Tubulin.
Cells were fixed and permeabilized. Then stained with the primary antibody (HA601187, 1μg/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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Flow cytometric analysis of NIH/3T3 cells labeling beta Tubulin.
Cells were fixed and permeabilized. Then stained with the primary antibody (HA601187, 1μg/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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Immunohistochemical analysis of paraffin-embedded human colon cancer tissue with Mouse anti-beta Tubulin antibody (HA601187) at 1/10,000 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 (HA601187) at 1/10,000 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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