Antibody data
- Antibody Data
- Antigen structure
- References [2]
- Comments [0]
- Validations
- Western blot [4]
- Immunocytochemistry [5]
- Immunohistochemistry [1]
- Other assay [1]
Submit
Validation data
Reference
Comment
Report error
- Product number
- MA1-848 - Provider product page
- Provider
- Invitrogen Antibodies
- Product name
- PLK1 Monoclonal Antibody (13E8)
- Antibody type
- Monoclonal
- Antigen
- Recombinant full-length protein
- Description
- MA1-848 detects PLK1 from human, mouse, and rat samples.
- Antibody clone number
- 13E8
- Concentration
- 1 mg/mL
Submitted references Mitotic entry upon Topo II catalytic inhibition is controlled by Chk1 and Plk1.
Small-molecular, non-peptide, non-ATP-competitive polo-like kinase 1 (Plk1) inhibitors with a terphenyl skeleton.
Arroyo M, Cañuelo A, Calahorra J, Hastert FD, Sánchez A, Clarke DJ, Marchal JA
The FEBS journal 2020 Nov;287(22):4933-4951
The FEBS journal 2020 Nov;287(22):4933-4951
Small-molecular, non-peptide, non-ATP-competitive polo-like kinase 1 (Plk1) inhibitors with a terphenyl skeleton.
Mita Y, Noguchi-Yachide T, Ishikawa M, Hashimoto Y
Bioorganic & medicinal chemistry 2013 Feb 1;21(3):608-17
Bioorganic & medicinal chemistry 2013 Feb 1;21(3):608-17
No comments: Submit comment
Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot analysis of PLK1 was performed by loading 25 µg of untreated or Nocodazole (100 nM, 48 hours) treated U2OS lysate from non-targeting control (Product # D-001810-10) or PLK1 SMART pool siRNA (Product # L-003290-00-0005) transfected U2OS cells onto a 4-20% Tris-HCl polyacrylamide gel. Proteins were transferred to a PVDF membrane and blocked with 5% Milk/TBST for at least 1 hour. Membranes were probed with PLK1 mouse monoclonal antibody (Product # MA1-848) at a dilution of 1:1000 overnight at 4°C on a rocking platform. Membranes were washed in TBS-0.1%Tween 20 and probed with a goat anti-mouse-HRP secondary antibody (Product # 31460) at a dilution of 1:20,000 for at least one hour. Membranes were washed and chemiluminescent detection performed using Pierce Super Signal West Dura (Product # 34075).
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot analysis of PLK1 was performed by loading 25 µg of untreated or Nocodazole (100 nM, 48 hours) treated U2OS lysate from non-targeting control (Product # D-001810-10) or PLK1 SMART pool siRNA (Product # L-003290-00-0005) transfected U2OS cells onto a 4-20% Tris-HCl polyacrylamide gel. Proteins were transferred to a PVDF membrane and blocked with 5% Milk/TBST for at least 1 hour. Membranes were probed with PLK1 mouse monoclonal antibody (Product # MA1-848) at a dilution of 1:1000 overnight at 4°C on a rocking platform. Membranes were washed in TBS-0.1%Tween 20 and probed with a goat anti-mouse-HRP secondary antibody (Product # 31460) at a dilution of 1:20,000 for at least one hour. Membranes were washed and chemiluminescent detection performed using Pierce Super Signal West Dura (Product # 34075).
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Knockdown of PLK1 was achieved by transfecting HeLa cells with PLK1 specific siRNAs (Silencer® select Product # s449 ). Western blot analysis (Fig a) was performed using Whole cell extract from the PLK1 knock down cells (lane 3), non-specific scrambled siRNA transfected cells (lane 2) and untransfected cells (lane 1). The blots were probed with Anti-PLK1 Mouse Monoclonal Antibody (Product # MA1-848, 1 µg/mL) and Goat anti-Mouse IgG (H+L) Superclonal™ Secondary Antibody, HRP conjugate (Product # A28177, 0.25 µg/mL, 1:4000 dilution). Densitometric analysis of this western blot is shown in histogram (Fig b). Loss of signal upon siRNA mediated knock down confirms that antibody is specific to PLK1.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot analysis was performed on whole cell extracts (30µg) of HeLa (Lane 1),HeLa treated with Nocodazole (500 nM for 18 hrs) (Lane2), SW-480 (Lane 3), SW-480 treated with Nocodazole (500 nM for 18 hrs) (Lane 4), COLO 205 (Lane 5), HCT 116 (Lane 6), K-562 (Lane 7), MDA-MB-231 (Lane 8), HepG2 (Lane 9), A549 (Lane 10). The blots were probed with PLK1 Mouse Monoclonal Antibody (Product# MA1-848, 1:1000 dilution) and detected by chemiluminescence using Goat anti-Mouse IgG (H+L) Superclonal™ Secondary Antibody, HRP conjugate (Product # A28177, 0.25µg/mL, 1:4000 dilution). A ~ 66 kDa band corresponding to PLK1 was observed across the cell lines tested. Known quantity of protein samples were electrophoresed using Novex® NuPAGE® 4-12 % Bis-Tris gel (Product # NP0321BOX), XCell SureLock™ Electrophoresis System (Product # EI0002) and Novex® Sharp Pre-Stained Protein Standard (Product # LC5800). Resolved proteins were then transferred onto a nitrocellulose membrane by wet transfer method. The membrane was probed with the relevant primary and secondary Antibody following blocking with 5 % skimmed milk. Chemiluminescent detection was performed using Pierce™ ECL Western Blotting Substrate (Product # 32106).
Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunofluorescent analysis of PLK1 using PLK1 Monoclonal antibody (13E8) (Product # MA1-848) shows staining in HeLa cells. PLK1 staining (green), F-Actin staining with Phalloidin (red) and nuclei with DAPI (blue) is shown. Cells were grown on chamber slides and fixed with formaldehyde prior to staining. Cells were probed without (control) or with or an antibody recognizing PLK1 (Product # MA1-848) at a dilution of 1:20 over night at 4 °C, washed with PBS and incubated with a DyLight-488 conjugated secondary antibody (Product # 35552 for GAR, Product # 35503 for GAM). Images were taken at 60X magnification.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunofluorescent analysis of PLK1 using PLK1 Monoclonal antibody (13E8) (Product # MA1-848) shows staining in U251 glioma cells. PLK1 staining (green), F-Actin staining with Phalloidin (red) and nuclei with DAPI (blue) is shown. Cells were grown on chamber slides and fixed with formaldehyde prior to staining. Cells were probed without (control) or with or an antibody recognizing PLK1 (Product # MA1-848) at a dilution of 1:20 over night at 4 °C, washed with PBS and incubated with a DyLight-488 conjugated secondary antibody (Product # 35552 for GAR, Product # 35503 for GAM). Images were taken at 60X magnification.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunofluorescent analysis of PLK1 using PLK1 Monoclonal antibody (13E8) (Product # MA1-848) shows staining in WiDr colon carcinoma cells. PLK1 staining (green), F-Actin staining with Phalloidin (red) and nuclei with DAPI (blue) is shown. Cells were grown on chamber slides and fixed with formaldehyde prior to staining. Cells were probed without (control) or with or an antibody recognizing PLK1 (Product # MA1-848) at a dilution of 1:20 over night at 4 °C, washed with PBS and incubated with a DyLight-488 conjugated secondary antibody (Product # 35552 for GAR, Product # 35503 for GAM). Images were taken at 60X magnification.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunofluorescence analysis of PLK1 was performed using 70% confluent log phase HeLa cells. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 10 minutes, and blocked with 1% BSA for 1 hour at room temperature. The cells were labeled with PLK1 Mouse monoclonal Antibody (Product # MA1-848) at 3µg/mL in 0.1% BSA and incubated for overnight at 4°C and then labeled with Goat anti-Mouse IgG (H+L) Superclonal™ Secondary Antibody, Alexa Fluor® 488 conjugate (Product # A28175) at a dilution of 1:2000 for 45 minutes at room temperature (Panel a: green). Nuclei (Panel b: blue) were stained with SlowFade® Gold Antifade Mountant with DAPI (Product # S36938). F-actin (Panel c: red) was stained with Rhodamine Phalloidin (Product # R415, 1:300). Panel d represents the merged image showing cytoplasmic and nuclear localization. Panel e shows the no primary antibody control. The images were captured at 60X magnification.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunofluorescence analysis of PLK1 was performed using 70% confluent log phase HeLa cells. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 10 minutes, and blocked with 1% BSA for 1 hour at room temperature. The cells were labeled with PLK1 Mouse monoclonal Antibody (Product # MA1-848) at 3µg/mL in 0.1% BSA and incubated for overnight at 4°C and then labeled with Goat anti-Mouse IgG (H+L) Superclonal™ Secondary Antibody, Alexa Fluor® 488 conjugate (Product # A28175) at a dilution of 1:2000 for 45 minutes at room temperature. Progression of cells through subsequent phases of the cell cycle results in differential localization of PLK1 in spindle poles, kinetochores and cleavage furrow. The images were captured at 60X magnification.
Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunohistochemistry was performed on normal deparaffinized human Colon tissue. To expose target proteins, heat induced antigen retrieval was performed using 10mM sodium citrate (pH6.0) buffer, microwaved for 8-15 minutes. Following antigen retrieval tissues were blocked in 3% BSA-PBS for 30 minutes at room temperature. Tissues were then probed at a dilution of 1:200 with a mouse monoclonal antibody recognizing PLK1 (Product # MA1-848) or without primary antibody (negative control) overnight at 4°C in a humidified chamber. Tissues were washed extensively with PBST and endogenous peroxidase activity was quenched with a peroxidase suppressor. Detection was performed using a biotin-conjugated secondary antibody and SA-HRP, followed by colorimetric detection using DAB. Tissues were counterstained with hematoxylin and prepped for mounting.
Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- 3 Fig. The ability of caffeine to bypass the G2 decatenation checkpoint is strongly perturbed if Plk1 function is depleted by RNAi. (A) Description of the experimental procedure performed. HeLa H2B-Red1 cells were synchronized at the G1/S border by double thymidine block. MCPH1 depletion and/or Plk1 depletion were achieved by transfection with siRNAs duplexes during the release from the first thymidine block. ICRF-193 (7 u m ) or solvent (DMSO) was added 7 h after release from the second thymidine block, while caffeine or solvent (medium) was added 2 h afterward. Time-lapse phase-contrast and fluorescent images were collected immediately after UCN01 with a Leica TCS SP5 microscope. Images were stacked and processed using imagej software. Timing data were obtained after visual inspection of mitosis onset, revealed by NEB, of 50 cells. (B) Immunoblot analysis of Plk1, MCPH1, and alpha-tubulin (loading control) levels in HeLa H2B-Red1 cells transfected with the indicated siRNAs as explained in (A). (C-F) Cumulative frequency chart showing the timing (in minutes) of mitosis onset, revealed by NEB, of HeLa H2B-Red1 cells after the corresponding treatments in the absence (C, E) or presence of caffeine (D, F) as explained in (A). (G) Immunoblot analysis of Plk1, MCPH1, and alpha-tubulin (loading control) levels in HeLa H2B-Red1 cells transfected with the indicated siRNAs. (H, I) Same analyses as in (C) and (D) but using a different siRNA oligo against Plk1.