Anti-Sec8 antibody [14G1]
| Name | Anti-Sec8 antibody [14G1] |
|---|---|
| Supplier | Abcam |
| Catalog | ab13254 |
| Prices | $370.00 |
| Sizes | 50 µg |
| Host | Mouse |
| Clonality | Monoclonal |
| Isotype | IgG1 |
| Clone | 14G1 |
| Applications | IP WB FC IHC-F |
| Species Reactivities | Mouse, Human, Rat, Sheep, Chicken, Bovine, Dog, Xenopus, Rainbow trout |
| Antigen | Fusion protein (Recombinant rat rSec8 (his-tagged) expressed in E |
| Description | Mouse Monoclonal |
| Gene | EXOC4 |
| Conjugate | Unconjugated |
| Supplier Page | Shop |
Product images
![All lanes : Anti-Sec8 antibody [14G1] (ab13254) at 5 µg/mlLane 1 : Brain (Human) Tissue Lysate - adult normal tissue (ab29466)Lane 2 : Kidney (Human) Tissue Lysate - adult normal tissue (ab30203)Lane 3 : Placenta (Human) Tissue Lysate - adult normal tissue (ab29745)Lane 4 : Brain (Mouse) Tissue Lysate Lysates/proteins at 20 µg per lane.SecondaryGoat polyclonal Secondary Antibody to Mouse IgG - H&L (HRP), pre-adsorbed at 1/5000 dilutiondeveloped using the ECL techniquePerformed under reducing conditions.](http://www.bioprodhub.com/system/product_images/ab_products/2/sub_4/27931_Sec8-Primary-antibodies-ab13254-2.jpg)
All lanes : Anti-Sec8 antibody [14G1] (ab13254) at 5 µg/mlLane 1 : Brain (Human) Tissue Lysate - adult normal tissue (ab29466)Lane 2 : Kidney (Human) Tissue Lysate - adult normal tissue (ab30203)Lane 3 : Placenta (Human) Tissue Lysate - adult normal tissue (ab29745)Lane 4 : Brain (Mouse) Tissue Lysate Lysates/proteins at 20 µg per lane.SecondaryGoat polyclonal Secondary Antibody to Mouse IgG - H&L (HRP), pre-adsorbed at 1/5000 dilutiondeveloped using the ECL techniquePerformed under reducing conditions.
![Overlay histogram showing HeLa cells stained with ab13254 (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 (ab13254, 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_4/27932_Sec8-Primary-antibodies-ab13254-3.jpg)
Overlay histogram showing HeLa cells stained with ab13254 (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 (ab13254, 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.
Immunofluorescence analysis of HMVECs, staining Sec8 with ab13254. Cells were fixed in paraformaldehyde, permeabilized with 0.2% Triton X-100 and blocked with blocking buffer. Samples were incubated with primary antibody (1/50) overnight at 4°C. An AlexFluor®-conjugated anti-mouse IgG (1/500) was used as a secondary antibody.
Sec8 was immunoprecipitated using 0.5mg Mouse Brain tissue lysate, 5µg of Mouse monoclonal to Sec8 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, Mouse Brain tissue lysate 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 ab13254.Secondary: Goat polyclonal to mouse IgG light chain specific (HRP) at 1/20,000 dilution.Band: 117kDa, non specific bands - 45kDa: We are unsure as to the identity of this extra band; Sec8
Product References
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Exocyst complex component 3-like 2 (EXOC3L2) associates with the exocyst complex - Exocyst complex component 3-like 2 (EXOC3L2) associates with the exocyst complex
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Dishevelled controls apical docking and planar polarization of basal bodies in - Dishevelled controls apical docking and planar polarization of basal bodies in
Park TJ, Mitchell BJ, Abitua PB, Kintner C, Wallingford JB. Nat Genet. 2008 Jul;40(7):871-9.