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The details of the immunogen for this antibody are not available.
Our Abpromise guarantee covers the use of ab6326 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|IHC (PFA fixed)||1/40.|
|ICC||Use at an assay dependent concentration.|
|IHC-Fr||1/40 - 1/200. PubMed: 16670699In addition, found to work at 1/400. For PFA fixed tissue use at 1/40, from PMID 16373695.|
|Flow Cyt||1/40. Ab18450-Rat monoclonal IgG2a, is suitable for use as an isotype control with this antibody.|
|IHC-FrFl||Use at an assay dependent concentration.|
ICC/IF image of ab6326 stained HeLa cells, both BrdU treated (left image) and normal cells (right image). The cells were 100% methanol fixed (5 min) and then subjected to acid hydrolysis using 2M HCL in 0.1% PBS-Tween for 30 minutes at room temperature to denature the DNA. They were 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 (ab6326, 10µg/ml) overnight at +4°C. The secondary antibody (green) was ab98420, DyLight® 488 goat anti-rat IgG (H+L) used at a 1/250 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. Positive staining can be seen in the BrdU treated cells, but not in the normal cells, demonstrating specificity for BrdU.
ab6326 used in double labeling on mouse sciatic nerve tissue (5
ab6326 staining mouse adult epidermis tissue sections by IHC-P. Sections were PFA fixed and subjected to heat mediated antigen retrieval in citrate buffer prior to blocking in 10% serum for 1 hr at RT. The primary antibody was diluted 1/200 and incubated with the sample for 24 hrs at 4°C. A biotinylated goat anti-rat antibody was used as the secondary followed by a streptavidin-Cy3®.
ab6326 staining cultured cells of rat brain tissue by ICC. The sample was PFA fixed and permeabilized in 1M HCl prior to blocking with 5% serum for 1 hour at 25°C. The primary antibody was diluted 1/500 and incubated with the sample for 16 hours at 25°C. A biotinylated rabbit anti-rat IgG antibody, diluted 1/200, was used as the secondary.
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