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Western blot analysis of TATA binding protein TBP on different lysates with Rabbit anti-TATA binding protein TBP antibody (HA724274) at 1/5,000 dilution.
Lane 1: HeLa cell lysate (15 µg/Lane)
Lane 2: 293T cell lysate (15 µg/Lane)
Lane 3: HCT 116 cell lysate (15 µg/Lane)
Lane 4: COS-1 cell lysate (15 µg/Lane)
Lane 5: C2C12 cell lysate (15 µg/Lane)
Lane 6: NIH/3T3 cell lysate (15 µg/Lane)
Lane 7: C6 cell lysate (15 µg/Lane)
Predicted band size: 38 kDa
Observed band size: 35-40 kDa
Exposure time: 20 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 (HA724274) at 1/5,000 dilution was used in primary antibody dilution (K1803) at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.
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Western blot analysis of TATA binding protein TBP on different lysates with Rabbit anti-TATA binding protein TBP antibody (HA724274) at 1/5,000 dilution.
Lane 1: Mouse testis tissue lysate
Lane 2: Mouse colon tissue lysate
Lane 3: Rat testis tissue lysate
Lysates/proteins at 20 µg/Lane.
Predicted band size: 35 kDa
Observed band size: 35 kDa
Exposure time: 20 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 (HA724274) at 1/5,000 dilution was used in primary antibody dilution (K1803) at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.
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Immunocytochemistry analysis of HeLa cells labeling TATA binding protein TBP with Rabbit anti-TATA binding protein TBP antibody (HA724274) at 1/250 dilution.
Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 15 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 Rabbit anti-TATA binding protein TBP antibody (HA724274) at 1/250 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) 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 (HA601187, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution.
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Immunocytochemistry analysis of NIH/3T3 cells labeling TATA binding protein TBP with Rabbit anti-TATA binding protein TBP antibody (HA724274) at 1/250 dilution.
Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 15 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 Rabbit anti-TATA binding protein TBP antibody (HA724274) at 1/250 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) 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 (HA601187, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution.
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Immunocytochemistry analysis of C6 cells labeling TATA binding protein TBP with Rabbit anti-TATA binding protein TBP antibody (HA724274) at 1/250 dilution.
Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 15 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 Rabbit anti-TATA binding protein TBP antibody (HA724274) at 1/250 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) 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 (HA601187, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution.
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Flow cytometric analysis of HeLa cells labeling TATA binding protein TBP.
Cells were fixed and permeabilized. Then stained with the primary antibody (HA724274, 1/1,000) (red) compared with Rabbit IgG 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-Rabbit IgG Secondary antibody (HA1121) 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 TATA binding protein TBP.
Cells were fixed and permeabilized. Then stained with the primary antibody (HA724274, 1/1,000) (red) compared with Rabbit IgG 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-Rabbit IgG Secondary antibody (HA1121) 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 C6 cells labeling TATA binding protein TBP.
Cells were fixed and permeabilized. Then stained with the primary antibody (HA724274, 1/1,000) (red) compared with Rabbit IgG 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-Rabbit IgG Secondary antibody (HA1121) 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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TATA binding protein TBP was immunoprecipitated from 0.2 mg HeLa cell lysate with HA724274 at 2 µg/10 µl beads. Western blot was performed from the immunoprecipitate using HA724274 at 1/5,000 dilution. HRP Conjugated Anti-Rabbit IgG for IP Nano-secondary antibody at 1/5,000 dilution was used for 1 hour at room temperature.
Lane 1: HeLa cell lysate (input)
Lane 2: HA724274 IP in HeLa cell lysate
Lane 3: Rabbit IgG instead of HA724274 in HeLa cell lysate
Blocking/Dilution buffer: primary antibody dilution (K1803)
Exposure time: 20 seconds; ECL: K1801
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