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Full length native protein (purified) (S. cerevisiae).
This antibody can be used as a loading control on Western blots (Allen et al.) and is not detected by anti-mouse Ig secondaries. It has been used in epitope tagging procedures to detect proteins tagged with a C-terminal Gly-Gly-Phe(OH) epitope. Under some circumstances this antibody may cross-react with other protein including E. coli rec A and oxidized actin.
Our Abpromise guarantee covers the use of ab6160 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|ELISA||Use at an assay dependent dilution.|
|IHC-Fr||Use at an assay dependent dilution.|
|IP||Use at an assay dependent dilution.|
|RIA||Use at an assay dependent dilution.|
|WB||1/5000 - 1/10000.|
|Flow Cyt||Use 1µg for 106 cells.|
|ICC/IF||1/1000. (see PMID: 16230461)|
|IHC (PFA fixed)||Use a concentration of 5 µg/ml. (from PubMed:16966421)|
|IHC-P||Use at an assay dependent dilution.|
|IHC (Methanol fixed)||1/100. (from PubMed:16943269).|
ICC/IF image of ab6160 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 ab6160 at 5µg/ml overnight at +4°C. The secondary antibody (pseudo-colored green) was Alexa Fluor® 488 goat anti- rat (ab150165) IgG (H+L) preadsorbed, used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (pseudo-colored red) at a 1/200 dilution for 1h at room temperature. DAPI was used to stain the cell nuclei (pseudo-colored blue) at a concentration of 1.43µM for 1hour at room temperature.
Western blot against tubulin with ab6160. Secondary Rabbit polyclonal to Rat IgG H&L (HRP) (ab6734) was used at 1/2000.
Exposure time: 10 seconds.
Lane 1-2: 20
Lane 3-4: 20
Lane 1,3: ab6160 at 1/5000.
Lane 2,4: ab6160 at 1/10000.
ab6160 staining Tubulin in mouse MEF cells by Immunocytochemistry/ Immunofluorescence. Cells were fixed with 2% PFA and 96% Ethanol. Samples were incubated with primary antibody (1/2000 in 0.1% Saponin/1% BSA/PBS) for 1 hour. A Cyt3®-conjugated goat polyclonal to rat IgG (H&L) was used at dilution at 1/500 as secondary antibody. Red staining in the image represents Tubulin, whereas the green one resembles gamma-tubulin.
ab6160 staining mouse prostate tissue sections by IHC-P. The tissue was serial sectioned at 6 microns, formaldehyde fixed and subjected to heat mediated antigen retrieval prior to blocking in 3% peroxidase for 5 minutes at 27°C. The primary antibody was diluted 1/500 and incubated with the sample for 16 hours at 4°C. A Cy5® conjugated goat anti-rat antibody was used as the secondary.
ab6160 at 1/1000 dilution staining Tubulin in human WBC cells by Immunocytochemistry/ Immunofluorescence. Cells were fixed in acetone and then blocked in 5% serum for 1 hour at 25°C. No permeabilization was done. The primary antibody was used at 1/1000 dilution in PBS-Tween and incubated with sample at 4°C for 16 hours. An Alexa Fluor® 594 conjugated goat polyclonal to rat IgG was used as secondary at 1/500 dilution.
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