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Recombinant full length protein (Human) from insect cells.
Please note the isotype for this product is IgM and requires the use of an anti-IgM secondary.
Our Abpromise guarantee covers the use of ab5181 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|Flow Cyt||Use 1-2µg for 106 cells. (Also see Abreview)|
|IHC-P||Use at an assay dependent concentration.|
|ICC/IF||Use at an assay dependent concentration. Fix with 1:1 ice cold MEOH:acetone and place at -20°C for 10 minutes.|
|WB||1/500 - 1/1000. Detects a band of approximately 127 kDa (predicted molecular weight: 127 kDa).
Please note the isotype for this product is IgM and requires the use of an anti-IgM secondary. Even though we have a positive Abreview on IHC, this is not a batch tested application and we cannot guarantee that it will work.
|IP||Use at an assay dependent concentration.|
Overlay histogram showing Jurkat cells stained with ab5181 (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 followed by the antibody (ab5181, 2µg/1x106 cells) for 30 min at 22ºC. The secondary antibody used was DyLight® 488 goat anti-mouse IgM (mu chain) (ab97007) at 1/500 dilution for 30 min at 22ºC. Isotype control antibody (black line) was mouse IgM [ICIGM] (ab91545, 2µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed.
ICC/IF image of ab5181 stained MCF-7 cells. The cells were 4% formaldehyde fixed (10 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 ab5181 at 1/200 dilution overnight at +4°C. The secondary antibody (pseudo-colored green) was Alexa Fluor® 488 goat anti-mouse IgM (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.
Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"