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Full length native protein (purified) (S. cerevisiae).
We can conjugate this antibody to FITC for you (please see ab150252 for details). This antibody can be used as a Western blotting loading control (Kops et al.) and as a Microtubule Marker.
Has been used for the selection of specific recombinant antibodies engineered to incorporate its epitope. It is also useful for studying the function of microtubules.
Our Abpromise guarantee covers the use of ab6161 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|WB||Use a concentration of 1 µg/ml.|
|IHC-FoFr||1/600. PubMed: 15831501Suggested working dilution of 1/600.|
|RIA||Use at an assay dependent concentration.|
|Flow Cyt||Use 1µg for 106 cells.
ab18450 - Rat monoclonal IgG2a, is suitable for use as an isotype control with this antibody.
|ICC/IF||Use a concentration of 5 µg/ml.|
Western blot against tubulin with ab6161 at 1/3000. Secondary Rabbit anti-Rat IgG HRP (ab6734)was used at 1/2000. Exposure time: 2mins.
Lane 1: 20
Lane 2: 20
Lane 3: 20
Confocal image of 21 day in vitro rat hippocampal neurons, stained with rat monoclonal antibody to Tubulin - Microtubule Marker (ab6161) in green at 1/500 and Microtubule Associated protein 2 in blue.
This picture was kindly supplied as part of the review submitted by Dr Jonathon Burman.
Cultured human macrophages were used with ab6161 at 1/1000 for immunofluorescence. Cells were fixed with cold 2% formaldehyde for 20mins.
Green staining is Alexa 568, Blue staining is DAPI stain.
This cell represents a young macrophage, the staining patterns varied as the cells aged in culture.
This image is courtesy of an anonymous Abreview
ICC/IF image of ab6161 stained human HepG2 cells. The cells were 4% PFA fixed (10 min), permabilised in 0.1% PBS-Tween (20 min) and incubated with the antibody (ab6161, 1µg/ml) for 1h at room temperature. 1%BSA / 10% normal goat serum / 0.3M glycine was used to block non-specific protein-protein interactions. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rat IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red). DAPI was used to stain the cell nuclei (blue). This antibody also gave a positive IF result in HeLa, HEK 293 and MCF7 cells.
ab6161 staining mouse NIH 3T3 fibroblast cells by ICC/IF. Cells were PFA fixed and permeabilized in 0.2% Triton X-100 prior to blocking in 5% BSA for 45 minutes at RT. The primary antibody was diluted 1/1000 and incubated with the sample for 1 hour. An Alexa Fluor® 568 conjugated goat anti-rat antibody, diluted 1/3000, was used as the secondary.
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