WB
Western blot analysis of PLK1 on different lysates with Rabbit anti-PLK1 antibody at 1/1,000 dilution. Lane 1: HeLa cell lysate (20 µg/Lane), Lane 2: K-562 cell lysate (20 µg/Lane), Lane 3: HEK-293 cell lysate (20 µg/Lane), Lane 4: Raji cell lysate (20 µg/Lane), Lane 5: HT-29 cell lysate (20 µg/Lane), Lane 6: HCT 116 cell lysate (20 µg/Lane), Lane 7: COS-1 cell lysate (20 µg/Lane), Lane 8: RAW264.7 cell lysate (20 µg/Lane), Lane 9: C6 cell lysate (20 µg/Lane), Lane 10: Mouse testis tissue lysate (40 µg/Lane), Lane 11: Rat testis tissue lysate (40 µg/Lane), Exposure time: 1 minute; 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 1/1,000 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.IHC
Immunohistochemical analysis of paraffin-embedded human colon cancer tissue with Rabbit anti-PLK1 antibody at 1/200 dilution. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for 2 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 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.ICC/IF
Immunocytochemistry analysis of RAW264.7 cells labeling PLK1 with Rabbit anti-PLK1 antibody 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 Rabbit anti-PLK1 antibody at 1/100 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.| Product Name | PLK1 Recombinant Rabbit Monoclonal Antibody |
|---|---|
| Antibody Type | Primary Antibodies |
| Immunogen | Recombinant protein within human PLK1 aa 314-603 / 603. |
| Clonality | monoclonal |
|---|---|
| Isotype | IgG |
| Host Species | Rabbit |
| Tested Applications | ICC/IFIHCWB |
| WB:1:1000 IHC:1:200 ICC/IF:1:100 |
|
| Species Reactivity | HumanMonkeyMouseRat |
| Concentration | 1mg/ml |
| Purification | Protein A |
| Gene Symbol | PLK1 |
|---|---|
| Gene Synonyms | PLK STPK13 |
| Gene Full Name | polo like kinase 1 |
| Gene Summary | The Ser/Thr protein kinase encoded by this gene belongs to the CDC5/Polo subfamily. It is highly expressed during mitosis and elevated levels are found in many different types of cancer. Depletion of this protein in cancer cells dramatically inhibited cell proliferation and induced apoptosis; hence, it is a target for cancer therapy. [provided by RefSeq, Sep 2015] |
| Molecular Weight(MW) | 68kDa |
| Cellular Localization | Nucleus, Chromosome, centromere, kinetochore, Cytoplasm, cytoskeleton, microtubule organizing center, spindle, Midbody. |
WB
Western blot analysis of PLK1 on different lysates with Rabbit anti-PLK1 antibody at 1/1,000 dilution. Lane 1: HeLa cell lysate (20 µg/Lane), Lane 2: K-562 cell lysate (20 µg/Lane), Lane 3: HEK-293 cell lysate (20 µg/Lane), Lane 4: Raji cell lysate (20 µg/Lane), Lane 5: HT-29 cell lysate (20 µg/Lane), Lane 6: HCT 116 cell lysate (20 µg/Lane), Lane 7: COS-1 cell lysate (20 µg/Lane), Lane 8: RAW264.7 cell lysate (20 µg/Lane), Lane 9: C6 cell lysate (20 µg/Lane), Lane 10: Mouse testis tissue lysate (40 µg/Lane), Lane 11: Rat testis tissue lysate (40 µg/Lane), Exposure time: 1 minute; 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 1/1,000 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.
IHC
Immunohistochemical analysis of paraffin-embedded human colon cancer tissue with Rabbit anti-PLK1 antibody at 1/200 dilution. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for 2 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 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.
ICC/IF
Immunocytochemistry analysis of RAW264.7 cells labeling PLK1 with Rabbit anti-PLK1 antibody 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 Rabbit anti-PLK1 antibody at 1/100 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.| Application Notes | WB:1:1000 IHC:1:200 ICC/IF:1:100 |
|---|
| Form | Liquid |
|---|---|
| Storage Instructions | Store at +4℃ after thawing. Aliquot store at -20℃. Avoid repeated freeze / thaw cycles. |
| Storage Buffer | 1*TBS (pH7.4), 0.05% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide. |
Data sheet for OM644099