WB
Western blot analysis of UHRF1 on different lysates with Rabbit anti-UHRF1 antibody 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. Exposure time: 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 at 1/2,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody at 1/50,000 dilution was used for 1 hour at room temperature.ICC/IF
Immunocytochemistry analysis of HCT 116 cells labeling UHRF1 with Rabbit anti-UHRF1 antibody 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 at 1/10,000 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (488) 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 (red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (594) was used as the secondary antibody at 1/1,000 dilution.FC
Flow cytometric analysis of HCT 116 cells labeling UHRF1. Cells were fixed and permeabilized. Then stained with the primary antibody (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 at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).IP
UHRF1 was immunoprecipitated from 0.2 mg HL-60 cell lysate with Rabbit anti-UHRF1 antibody at 2 µg/10 µl beads. Western blot was performed from the immunoprecipitate using Rabbit anti-UHRF1 antibody 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:Rabbit anti-UHRF1 antibody IP in HL-60 cell lysate, Lane 3: Rabbit IgG instead of Rabbit anti-UHRF1 antibody in HL-60 cell lysate, Blocking/Dilution buffer: 5% NFDM/TBST, Exposure time: 9 seconds.| Product Name | UHRF1 Recombinant Rabbit Monoclonal Antibody |
|---|---|
| Antibody Type | Primary Antibodies |
| Immunogen | Recombinant protein within human UHRF1 aa 1-350. |
| Clonality | monoclonal |
|---|---|
| Isotype | IgG |
| Host Species | Rabbit |
| Tested Applications | FCICC/IFIPWB |
| WB:1:2000 ICC/IF:1:10000 FC:1:1000 IP:1-2μg/sample |
|
| Species Reactivity | HumanMonkeyMouse |
| Concentration | 1mg/ml |
| Purification | Protein A |
| Gene Symbol | UHRF1 |
|---|---|
| Gene Synonyms | Np95 hNP95 ICBP90 RNF106 TDRD22 hUHRF1 huNp95 |
| Gene Full Name | ubiquitin like with PHD and ring finger domains 1 |
| Gene Summary | This gene encodes a member of a subfamily of RING-finger type E3 ubiquitin ligases. The protein binds to specific DNA sequences, and recruits a histone deacetylase to regulate gene expression. Its expression peaks at late G1 phase and continues during G2 and M phases of the cell cycle. It plays a major role in the G1/S transition by regulating topoisomerase IIalpha and retinoblastoma gene expression, and functions in the p53-dependent DNA damage checkpoint. It is regarded as a hub protein for the integration of epigenetic information. This gene is up-regulated in various cancers, and it is therefore considered to be a therapeutic target. Multiple transcript variants encoding different isoforms have been found for this gene. A related pseudogene exists on chromosome 12. [provided by RefSeq, Feb 2014] |
| Alternative Names | Ac2-121 antibody AL022808 antibody E3 ubiquitin-protein ligase UHRF1 antibody EC 6.3.2.- antibody FLJ21925 antibody hNP95 antibody hUHRF1 antibody HuNp95 antibody ICBP90 antibody Inverted CCAAT box binding protein of 90 kDa antibody Ac2-121 antibody AL022808 antibody E3 ubiquitin-protein ligase UHRF1 antibody EC 6.3.2.- antibody FLJ21925 antibody hNP95 antibody hUHRF1 antibody HuNp95 antibody ICBP90 antibody Inverted CCAAT box binding protein of 90 kDa antibody Inverted CCAAT box binding protein 90-kD antibody Inverted CCAAT box-binding protein of 90 kDa antibody Liver regeneration-related protein LRRG126 antibody MGC138707 antibody NP95 antibody Nuclear phosphoprotein 95 antibody Nuclear phosphoprotein 95-KD antibody Nuclear protein 95 antibody Nuclear zinc finger protein Np95 antibody RING finger protein 106 antibody RNF106 antibody TDRD22 antibody Transcription factor ICBP90 antibody Ubiquitin like containing PHD and RING finger domains protein 1 antibody Ubiquitin like PHD and RING finger domain containing protein 1 antibody Ubiquitin-like PHD and RING finger domain-containing protein 1 antibody Ubiquitin-like protein containing PHD and RING finger domains 1 antibody Ubiquitin-like with PHD and ring finger domains 1 antibody Ubiquitin-like containing PHD and RING finger domains 1 antibody Ubiquitin-like-containing PHD and RING finger domains protein 1 antibody UHRF1 antibody UHRF1_HUMAN antibody |
| Molecular Weight(MW) | 90kDa(Observed band size: 90-100kDa) |
| Cellular Localization | Nucleus. |
WB
Western blot analysis of UHRF1 on different lysates with Rabbit anti-UHRF1 antibody 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. Exposure time: 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 at 1/2,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody at 1/50,000 dilution was used for 1 hour at room temperature.
ICC/IF
Immunocytochemistry analysis of HCT 116 cells labeling UHRF1 with Rabbit anti-UHRF1 antibody 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 at 1/10,000 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (488) 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 (red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (594) was used as the secondary antibody at 1/1,000 dilution.
FC
Flow cytometric analysis of HCT 116 cells labeling UHRF1. Cells were fixed and permeabilized. Then stained with the primary antibody (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 at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).
IP
UHRF1 was immunoprecipitated from 0.2 mg HL-60 cell lysate with Rabbit anti-UHRF1 antibody at 2 µg/10 µl beads. Western blot was performed from the immunoprecipitate using Rabbit anti-UHRF1 antibody 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:Rabbit anti-UHRF1 antibody IP in HL-60 cell lysate, Lane 3: Rabbit IgG instead of Rabbit anti-UHRF1 antibody in HL-60 cell lysate, Blocking/Dilution buffer: 5% NFDM/TBST, Exposure time: 9 seconds.| Application Notes | WB:1:2000 ICC/IF:1:10000 FC:1:1000 IP:1-2μg/sample |
|---|
| Form | Liquid |
|---|---|
| Storage Instructions | Store at +4℃ after thawing. Aliquot store at -20℃. Avoid repeated freeze / thaw cycles. |
| Storage Buffer | PBS (pH7.4), 0.1% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide. |
