Mouse, Human 交差が予測される動物種:
Rat, Sheep, Chicken, Cow, Dog, Xenopus laevis, Rainbow trout
Fusion protein (Recombinant rat rSec8 (his-tagged) expressed in E. coli).
In Western Blot, this antibody gave a positive signal in the following tissue lysates: human brain; human kidney; human placenta; mouse brain.
Abcam is committed to meeting high standards of ethical manufacturing and has decided to discontinue this product by July 2018 as it has been generated by the ascites method. We are sorry for any inconvenience this may cause.
Shipped at 4°C. Upon delivery aliquot and store at -20°C. Avoid freeze / thaw cycles.
All lanes : Anti-Sec8 antibody [14G1] (ab13254) at 5 µg/ml
Lane 1 : Human brain tissue lysate - total protein (ab29466) Lane 2 : Human kidney tissue lysate - total protein (ab30203) Lane 3 : Human placenta tissue lysate - total protein (ab29745) Lane 4 : Brain (Mouse) Tissue Lysate
Lysates/proteins at 20 µg per lane.
Secondary All lanes : Goat polyclonal Secondary Antibody to Mouse IgG - H&L (HRP), pre-adsorbed at 1/5000 dilution
Developed using the ECL technique.
Performed under reducing conditions.
Predicted band size: 117 kDa Observed band size: 117 kDa Additional bands at: 45 kDa, 52 kDa. We are unsure as to the identity of these extra bands.
Exposure time: 8 minutes
Flow Cytometry-Anti-Sec8 antibody [14G1](ab13254)
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.
Immunocytochemistry/ Immunofluorescence - Anti-Sec8 antibody [14G1] (ab13254)Image from Barkefors I et al., J Biol Chem. 2011 Jul 8;286(27):24189-99. Epub 2011 May 12. Fig 4.; doi: 10.1074/jbc.M110.212209; July 8, 2011 The Journal of Biological Chemistry, 286, 24189-24199.
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
Winkelmann A et al. Changes in neural network homeostasis trigger neuropsychiatric symptoms. J Clin Invest124:696-711 (2014).
Read more (PubMed: 24430185) »
Barkefors I et al. Exocyst Complex Component 3-like 2 (EXOC3L2) Associates with the Exocyst Complex and Mediates Directional Migration of Endothelial Cells. J Biol Chem286:24189-99 (2011).
Read more (PubMed: 21566143) »