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Recombinant fragment corresponding to Human ATF6 aa 1-273. This monoclonal antibody was made against a partial protein containing amino acids 1-273 of human ATF6.
MGEPAGVAGTMESPFSPGLFHRLDEDWDSALFAELGYFTDTDELQLEAAN ETYENNFDNLDFDLDLMPWESDIWDINNQICTVKDIKAEPQPLSPASSSY SVSSPRSVDSYSSTQHVPEELDLSSSSQMSPLSLYGENSNSLSSAEPLKE DKPVTGPRNKTENGLTPKKKIQVNSKPSIQPKPLLLPAAPKTQTNSSVPA KTIIIQTVPTLMPLAKQQPIISLQPAPTKGQTVLLSQPTVVQLQAPGVLP SAQPVLAVAGGVTQLPNHVVNVVPAPSANSPVNGKLSVTKPVLQSTMRNV GSDIAVLRRQQRMIKNRESACQSRKKKKEYMLG
Our Abpromise guarantee covers the use of ab11909 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|ICC/IF||Use at an assay dependent concentration.|
|IHC-P||Use at an assay dependent concentration.|
|IHC-Fr||Use at an assay dependent concentration.|
|IP||Use at an assay dependent concentration.|
|ChIP||Use at an assay dependent concentration.|
|Flow Cyt||Use 1µg for 106 cells.
ab170190 - Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody.
|WB||Use a concentration of 1 - 5 µg/ml. Detects a band of approximately 90 kDa (predicted molecular weight: 75 kDa). Expected band sizes of endogenous protein between 66-97 kDa, transfected full length found at 97 kDa. Additional bands at 35/55 kDa have been observed which may correspond to cleavage products.|
Lane 1: Wild type HAP1 whole cell lysate (50 µg)
Lane 2: ATF6 knockout HAP1 whole cell lysate (50 µg)
Lanes 1 - 2: Merged signal (red and green). Green - ab11909 observed at 95 kDa. Red - loading control, ab181602, observed at 37 kDa.
ab11909 was shown not to recognize ATF6 when ATF6 knockout samples were used, along with additional cross-reactive bands. Wild-type and ATF6 knockout samples were subjected to SDS-PAGE. ab11909 and ab181602 (Rabbit anti GAPDH loading control) were incubated overnight at 4°C at 1/1000 dilution and 1/10000 dilution respectively. Blots were developed with Goat anti-Mouse IgG H&L (IRDye® 800CW) preabsorbed ab216772 and Goat anti-Rabbit IgG H&L (IRDye® 680RD) preabsorbed ab216777 secondary antibodies at 1/10000 dilution for 1 hour at room temperature before imaging.
ICC/IF image of ab11909 stained MCF7 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 (ab11909, 10µg/ml) overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 goat anti-mouse IgG (H+L) ab150113) 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.
ab11909 (3ug/ml) staining of mouse ATF6 in NIH3T3 cell lysate by Western Blot. The ~36 kDa observed band has not been characterized; it may be an ATF6 breakdown/cleavage product.
Overlay histogram showing HeLa cells stained with ab11909 (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. The cells were then incubated with the antibody (ab11909, 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 in 0.1% PBS-Tween used under the same conditions.
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