Anti-Unrip antibody [3G6]
| Name | Anti-Unrip antibody [3G6] |
|---|---|
| Supplier | Abcam |
| Catalog | ab46784 |
| Prices | $359.00 |
| Sizes | 100 µg |
| Host | Mouse |
| Clonality | Monoclonal |
| Isotype | IgG1 |
| Clone | 3G6 |
| Applications | ICC/IF ICC/IF FC IP WB IHC-P |
| Species Reactivities | Human |
| Antigen | Recombinant fusion protein (Human) |
| Description | Mouse Monoclonal |
| Gene | STRAP |
| Conjugate | Unconjugated |
| Supplier Page | Shop |
Product images
![All lanes : Anti-Unrip antibody [3G6] (ab46784) at 5 µg/mlLane 1 : Ramos (Human Burkitt's lymphoma cell line) Whole Cell LysateLane 2 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate Lane 3 : SW480 (Human colon adenocarcinoma cell line) Whole Cell Lysate Lysates/proteins at 10 µg per lane.SecondaryGoat polyclonal to Mouse IgG - H&L - Pre-Adsorbed (HRP) at 1/3000 dilution](http://www.bioprodhub.com/system/product_images/ab_products/2/sub_5/18158_Unrip-Primary-antibodies-ab46784-2.jpg)
All lanes : Anti-Unrip antibody [3G6] (ab46784) at 5 µg/mlLane 1 : Ramos (Human Burkitt's lymphoma cell line) Whole Cell LysateLane 2 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate Lane 3 : SW480 (Human colon adenocarcinoma cell line) Whole Cell Lysate Lysates/proteins at 10 µg per lane.SecondaryGoat polyclonal to Mouse IgG - H&L - Pre-Adsorbed (HRP) at 1/3000 dilution
IHC image of ab46784 staining in human colon cancer 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 ab46784, 1µg/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.
ICC/IF image of ab46784 stained HeLa cells. The cells were 100% methanol fixed (5 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 (ab46784, 5µg/ml) overnight at +4°C. The secondary antibody (green) was ab96879, DyLight® 488 goat anti-mouse IgG (H+L) used at a 1/250 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.
![Unrip was immunoprecipitated using 0.5mg Hela whole cell extract, 5µg of Mouse monoclonal [3G6] to Unrip (ab46784) and 50µl of protein G magnetic beads (+). No antibody was added to the control (-). The antibody was incubated under agitation with Protein G beads for 10min, Hela whole cell extract lysate diluted in RIPA buffer was added to each sample and incubated for a further 10min under agitation.Proteins were eluted by addition of 40µl SDS loading buffer and incubated for 10min at 70oC; 10µl of each sample was separated on a SDS PAGE gel, transferred to a nitrocellulose membrane, blocked with 5% BSA and probed with ab46784.Secondary: Goat polyclonal to mouse IgG light chain specific (HRP) at 1/5000 dilution.Band: 42kDa: Unrip.](http://www.bioprodhub.com/system/product_images/ab_products/2/sub_5/18161_Unrip-Primary-antibodies-ab46784-7.jpg)
Unrip was immunoprecipitated using 0.5mg Hela whole cell extract, 5µg of Mouse monoclonal [3G6] to Unrip (ab46784) and 50µl of protein G magnetic beads (+). No antibody was added to the control (-). The antibody was incubated under agitation with Protein G beads for 10min, Hela whole cell extract lysate diluted in RIPA buffer was added to each sample and incubated for a further 10min under agitation.Proteins were eluted by addition of 40µl SDS loading buffer and incubated for 10min at 70oC; 10µl of each sample was separated on a SDS PAGE gel, transferred to a nitrocellulose membrane, blocked with 5% BSA and probed with ab46784.Secondary: Goat polyclonal to mouse IgG light chain specific (HRP) at 1/5000 dilution.Band: 42kDa: Unrip.
![Overlay histogram showing HeLa cells stained with ab46784 (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 (ab46784, 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 HeLa cells fixed with 4% paraformaldehyde (10 min)/permeabilized with 0.1% PBS-Tween for 20 min used under the same conditions.](http://www.bioprodhub.com/system/product_images/ab_products/2/sub_5/18162_Unrip-Primary-antibodies-ab46784-6.jpg)
Overlay histogram showing HeLa cells stained with ab46784 (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 (ab46784, 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 HeLa cells fixed with 4% paraformaldehyde (10 min)/permeabilized with 0.1% PBS-Tween for 20 min used under the same conditions.
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