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Synthetic peptide (the amino acid sequence is considered to be commercially sensitive) (N terminal)
Produced using Abcam’s RabMAb® technology. RabMAb® technology is covered by the following U.S. Patents, No. 5,675,063 and/or 7,429,487.
A 40 μl trial size is available to purchase for this antibody.
The Alexa Fluor® 488 variant is available, see ab185031.
We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated ‘PUR’ on our product labels.
If you have any questions regarding this update, please contact our Scientific Support team.
Our Abpromise guarantee covers the use of ab52866 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|ICC/IF||1/250 - 1/500.|
|IHC - Wholemount||Use at an assay dependent concentration.|
|WB||1/1000 - 1/50000. Detects a band of approximately 50 kDa (predicted molecular weight: 50 kDa).|
|Flow Cyt||1/20 - 1/50.|
|IHC-P||Use at an assay dependent dilution.|
|IHC-Fr||Use at an assay dependent concentration. PubMed: 21933451|
alpha Tubulin was immunoprecipitated from 1mg of HeLa whole cell extract using ab52866 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab52866 at 1/1000 dilution. Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG, was used as secondary antibody at 1/1500 dilution.
Lane 1 Hela whole cell extract, Lane 2 PBS instead of whole cell extract.
Blocking and dilution buffer and concentration: 5% NFDM/TBST.
Flow cytometry analysis of 2% paraformaldehyde fixed HepG2 cells labeling alpha Tubulin with ab52866 at 1/130 dilution (red line). Secondary antibody used is a goat anti rabbit IgG (FITC) at 1/150 dilution. The isotype control is rabbit monoclonal IgG (black line). The unlabeled control is cells without incubation with primary and secondary antibodies (blue line).
Immunofluorescent analysis of 4% paraformaldehyde-fixed HeLa cells labeling alpha Tubulin with ab52866 at 1/500 dilution. The cells were permeabilised with 0.1% Triton X-100. Anti-rabbit Alexa Fluor® 488 (ab150077) at 1/400 dilution was used as the secondary antibody (green). The confocal image shows microtubules staining on HeLa cell line. The nuclear counter stain is DAPI (blue). Tubulin is detected with ab7291 (anti-Tubulin mouse mAb) at 1/500 and anti-mouse AlexaFluor® 594 (ab150120) at 1/500 dilution (red).
The negative controls are as follows:
1. ab52866 at 1/500 dilution followed by anti-mouse AlexaFluor® 594 (ab150120) at 1/500 dilution.
2. ab7291 (anti-Tubulin mouse mAb) at 1/500 dilution followed by anti-rabbit Alexa Fluor® 488 (ab150077) at 1/400 dilution.
Immunohistochemistry analysis of paraffin-embedded Pig kidney tissue labeling alpha Tubulin with ab52866 at a 1/1000 dilution. Cytoplasmic staining on Pig kidney tubule and weak on glomerulus shown. Anti-Rabbit HRP (ab97051) used at a 1/100 dilution. Counter stained with Hematoxylin.
Immunohistochemistry analysis of paraffin-embedded Rat kidney tissue labeling alpha Tubulin with ab52866 at a 1/1000 dilution. Cytoplasmic staining on Rat kidney tubule and weak on glomerulus shown. Secondary antibody Anti-Rabbit HRP (ab97051) used at a 1/500 dilution. Counter stained with Hematoxylin.
Immunohistochemistry analysis of paraffin-embedded Mouse kidney tissue labeling alpha Tubulin with ab52866 at a 1/1000 dilution. Cytoplasmic staining on Mouse kidney tubule shown. Secondary antibody Anti-Rabbit HRP (ab97051) used at a 1/500 dilution. Counter stained with Hematoxylin.
Immunohistochemistry analysis of paraffin-embedded Human breast cancer labeling alpha Tubulin with ab52866 at a 1/1000 dilution. Cytoplasmic staining on cancer cells shown. Secondary antibody ab97051 Goat Anti-Rabbit IgG H&L (HRP) used at a 1/500 dilution. Counter stained with Hematoxylin.
Inset image: negative control obtained using PBS instead of ab52866, secondary antibody is Anti-Rabbit HRP (ab97051) at 1/500 dilution.
This blot was produced using a 4-12% Bis-tris gel under the MOPS buffer system. The gel was run at 200V for 50 minutes before being transferred onto a Nitrocellulose membrane at 30V for 70 minutes. The membrane was then blocked for an hour using Licor blocking buffer before being incubated with ab52866 overnight at 4°C. Antibody binding was detected using Anti-Rabbit Alexa Fluor® 790 (ab175781) at a 1:10,000 dilution for 1hr at room temperature and then imaged using the Licor Odyssey CLx.
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