Anti-HEF1 antibody [2G9]
| Name | Anti-HEF1 antibody [2G9] |
|---|---|
| Supplier | Abcam |
| Catalog | ab18056 |
| Prices | $403.00 |
| Sizes | 100 µl |
| Host | Mouse |
| Clonality | Monoclonal |
| Isotype | IgG1 |
| Clone | 2G9 |
| Applications | WB IP ICC/IF ICC/IF IHC-F IHC-P FC |
| Species Reactivities | Mouse, Rat, Human |
| Antigen | Fusion protein, corresponding to amino acids 82-398 of Human HEF1 |
| Description | Mouse Monoclonal |
| Gene | NEDD9 |
| Conjugate | Unconjugated |
| Supplier Page | Shop |
Product images
ICC/IF image of ab18056 stained Mcf7 cells. The cells were 4% formaldehyde fixed (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab18056, 1µg/ml) overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 goat anti-mouse IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.
![Anti-HEF1 antibody [2G9] (ab18056) at 1/2000 dilution + whole cell lysate prepared from a murine neural stem cell line at 15 µgSecondaryHRP conjugated anti mouse IgG (H+L) at 1/3000 dilutiondeveloped using the ECL technique](http://www.bioprodhub.com/system/product_images/ab_products/2/sub_2/29769_HEF1-Primary-antibodies-ab18056-6.jpg)
Anti-HEF1 antibody [2G9] (ab18056) at 1/2000 dilution + whole cell lysate prepared from a murine neural stem cell line at 15 µgSecondaryHRP conjugated anti mouse IgG (H+L) at 1/3000 dilutiondeveloped using the ECL technique
![Overlay histogram showing A549 cells stained with ab18056 (red line). The cells were fixed with 80% 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 (ab18056, 1µg/1x106 cells) for 30 min at 22ºC. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22ºC. 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. This antibody gave a positive signal in A549 cells fixed with 4% paraformaldehyde (10 min)/permeabilized in 0.1% PBS-Tween used under the same conditions.](http://www.bioprodhub.com/system/product_images/ab_products/2/sub_2/29770_HEF1-Primary-antibodies-ab18056-7.jpg)
Overlay histogram showing A549 cells stained with ab18056 (red line). The cells were fixed with 80% 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 (ab18056, 1µg/1x106 cells) for 30 min at 22ºC. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22ºC. 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. This antibody gave a positive signal in A549 cells fixed with 4% paraformaldehyde (10 min)/permeabilized in 0.1% PBS-Tween used under the same conditions.
IHC image of ab18056 staining in human kidney formalin fixed paraffin embedded tissue section, performed on a Leica BondTM system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab18056, 10µg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.
Product References
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