Anti-CENPE antibody [1H12]
| Name | Anti-CENPE antibody [1H12] |
|---|---|
| Supplier | Abcam |
| Catalog | ab5093 |
| Prices | $401.00 |
| Sizes | 100 µl |
| Host | Mouse |
| Clonality | Monoclonal |
| Isotype | IgG1 |
| Clone | 1H12 |
| Applications | ICC/IF ICC/IF IP ICC/IF WB FC |
| Species Reactivities | Human |
| Antigen | Recombinant full length protein (Human) |
| Description | Mouse Monoclonal |
| Gene | CENPE |
| Conjugate | Unconjugated |
| Supplier Page | Shop |
Product images
![Kinetochores specific staining of HCT116 cells arrested in G2/M phase by nocodazole treatment. Methanol fixed cells were stained using mouse monoclonal [1H12] antibody to CENP-E ab5093 (green) and DAPI (blue). This image was kindly supplied as part of the review submitted by Salvador Rodrigez-Nieto.](http://www.bioprodhub.com/system/product_images/ab_products/2/sub_1/27575_ab5093_1.jpg)
Kinetochores specific staining of HCT116 cells arrested in G2/M phase by nocodazole treatment. Methanol fixed cells were stained using mouse monoclonal [1H12] antibody to CENP-E ab5093 (green) and DAPI (blue). This image was kindly supplied as part of the review submitted by Salvador Rodrigez-Nieto.
ab5093 at 1/500 staining human fibrosarcoma (HT1080) cells by ICC/IF. The cells were treated with 0.1-0.2ug/mL colcemid for 45-60 minutes, then swollen in hypotonic buffer for 8 minutes and centrifuged onto glass slides. Cells were blocked in 1X PBS + 1% BSA + 0.5% Triton X-100 (blocking buffer) for 30 minutes at room temperature. The antibodies were diluted 1/300-1/500 in blocking buffer and incubated overnight at 4 degrees C. ab5093 was detected with Alexa Fluor 488-donkey anti-mouse for 1-2 hours at room temperature.See Abreview
HeLa cells were stained with ab5093, anti-CENPE (in green) in panel one, and with ab5093 and SH-CREST (red), which stains the centromeres, in panel 2.Fix cells for 30 minutes on ice in 4% formaldehyde in PEM. Quench autofluorescence 2 x 5 minutes with 1 mg/ml Na borohydride or 100 mM ammonium chloride in PEM. Permeablize 30 minutes with 0.5% TX-100 in PEM. Block 30 minutes in 5% milk in TBST. Primary antibody overnight at 4oC diluted 1/250 in 5% milk in TBST. Secondary antibody was incubated for 1 hour at RT diluted in 5% milk in TBST. Post-fix 20 minutes on ice in 4% formaldehyde in PEM. Quench autofluorescence 2 x 5 minutes with ammonium chloride in PEM. Counterstain with DAPI in TBST. Mount with ProLong Gold antifade reagent from Invitrogen. Notes: Ample washing between each step. TBST = Tris buffered saline + 0.1% Tween. PEM = 80 mM K-PIPES, pH 6.8, 5 mM EGTA, 2 mM MgCl2.
![All lanes : Anti-CENPE antibody [1H12] (ab5093) at 1 µg/mlLane 1 : HeLa (Human epithelial carcinoma cell line) Whole Cell LysateLane 2 : HepG2 (Human hepatocellular liver carcinoma cell line) Whole Cell Lysate Lane 3 : HEK293 (Human embryonic kidney cell line) Whole Cell LysateLysates/proteins at 10 µg per lane.SecondaryGoat Anti-Mouse IgG H&L (HRP) preadsorbed (ab97040) at 1/5000 dilutiondeveloped using the ECL techniquePerformed under reducing conditions.](http://www.bioprodhub.com/system/product_images/ab_products/2/sub_1/27578_CENPE-Primary-antibodies-ab5093-2.jpg)
All lanes : Anti-CENPE antibody [1H12] (ab5093) at 1 µg/mlLane 1 : HeLa (Human epithelial carcinoma cell line) Whole Cell LysateLane 2 : HepG2 (Human hepatocellular liver carcinoma cell line) Whole Cell Lysate Lane 3 : HEK293 (Human embryonic kidney cell line) Whole Cell LysateLysates/proteins at 10 µg per lane.SecondaryGoat Anti-Mouse IgG H&L (HRP) preadsorbed (ab97040) at 1/5000 dilutiondeveloped using the ECL techniquePerformed under reducing conditions.
![Overlay histogram showing HeLA cells stained with ab5093 (red line). The cells were fixed with 100% methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab5093, 1µg/1x106 cells) for 30 min at 22°C. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22°C. Isotype control antibody (black line) was Mouse IgG1 [ICIGG1] (ab91353, 2µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed.](http://www.bioprodhub.com/system/product_images/ab_products/2/sub_1/27579_CENPE-Primary-antibodies-ab5093-3.jpg)
Overlay histogram showing HeLA cells stained with ab5093 (red line). The cells were fixed with 100% methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab5093, 1µg/1x106 cells) for 30 min at 22°C. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22°C. Isotype control antibody (black line) was Mouse IgG1 [ICIGG1] (ab91353, 2µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed.
Product References
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Glycogen synthase kinase 3beta interacts with and phosphorylates the - Glycogen synthase kinase 3beta interacts with and phosphorylates the
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