UHRF1 Recombinant Rabbit Monoclonal Antibody [PSH10-63]

Western blot analysis of UHRF1 on different lysates with Rabbit anti-UHRF1 antibody (HA723228) at 1/2,000 dilution.

Lane 1: HCT 116 cell lysate
Lane 2: HL-60 cell lysate
Lane 3: HepG2 cell lysate
Lane 4: Jurkat cell lysate
Lane 5: F9 cell lysate
Lane 6: NIH/3T3 cell lysate
Lane 7: COS-1 cell lysate

Lysates/proteins at 20 µg/Lane.

Predicted band size: 90 kDa
Observed band size: 90-100 kDa

Exposure time: 10 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 (HA723228) at 1/2,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.

Immunocytochemistry analysis of HCT 116 cells labeling UHRF1 with Rabbit anti-UHRF1 antibody (HA723228) at 1/10,000 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-UHRF1 antibody (HA723228) at 1/10,000 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.

Immunocytochemistry analysis of HL-60 cells labeling UHRF1 with Rabbit anti-UHRF1 antibody (HA723228) at 1/10,000 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-UHRF1 antibody (HA723228) at 1/10,000 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.

Flow cytometric analysis of HCT 116 cells labeling UHRF1.

Cells were fixed and permeabilized. Then stained with the primary antibody (HA723228, 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).

UHRF1 was immunoprecipitated from 0.2 mg HL-60 cell lysate with HA723228 at 2 µg/10 µl beads. Western blot was performed from the immunoprecipitate using HA723228 at 1/1,000 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: HL-60 cell lysate (input)
Lane 2: HA723228 IP in HL-60 cell lysate
Lane 3: Rabbit IgG instead of HA723228 in HL-60 cell lysate

Blocking/Dilution buffer: 5% NFDM/TBST
Exposure time: 9 seconds; ECL: K1801

Chromatin immunoprecipitations were performed with cross-linked chromatin from HCT 116 cells with UHRF1 (HA723228) or Normal Rabbit IgG according to the ChIP protocol. The enriched DNA was quantified by real-time PCR using indicated primers. The amount of immunoprecipitated DNA in each sample is represented as signal relative to the total amount of input chromatin, which is equivalent to one.