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The hybridoma was formed by the fusion of mouse myeloma NS1 cellswith spleen cells from rats immunized with B10 mouse spleen cells enrichedfor T lymphocytes.
CD11b is also known as MAC-1alpha, CR3 and integrin alpha M. We do not batch test ab8878 in IHC, however, some customers have successfully used this antibody in this application. We cannot recommend this antibody for IHC.
Our Abpromise guarantee covers the use of ab8878 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|ICC/IF||Use a concentration of 5 µg/ml.|
|IF||Use at an assay dependent concentration. PubMed: 28138565|
|Flow Cyt||Use at an assay dependent concentration.
Each labeling step was carried out for 30 min at 0-4°C in mediumcontaining 0.01 M NaN3. Cells (5 x 107/ml) were incubated with 50 µl M1 monoclonal antibody or irrelevant monoclonal antibody, R4/18.2, as control in the first step, washed, suspended in 50 µl of FITC-F (ab')2 anti-rat IgGin the second step, and washed through a layer of fetal calf serum. Can also be used in cytotoxicity and binding assays.
ab18536 - Rat monoclonal IgG2b, is suitable for use as an isotype control with this antibody.
ICC/IF image of ab8878 stained Raw 246.7 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 ab8878 at 5µg/ml overnight at +4°C. The secondary antibody (green) was DyLight® 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) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.
Overlay histogram showing Human macrophages stained with ab8878 (blue line). Cells were incubated with human immunoglobulin for 30 min at 4°C to block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab8878, 20 µg/mL) for 30 min at 4ºC. The secondary antibody used was a mouse anti-rat FITC (IgG2b gamma chain) (ab99671) at 1 µg/mL for 30 min at 4ºC. Isotype control antibody (black line) was Rat IgG2b [RTK4530] (ab18541) used under the same conditions.
Overlay histogram showing Human macrophages stained with ab8878 (red line). Cells were pre-incubated in Human AB serum (10%) for 20 mins at 4ºC. Cells were then incubated with the antibody (ab8878, 0.1µg/1x10^6 cells) for 30 min at 4ºC. The secondary antibody used was a goat anti-rat Alexa Fluor® 488 (IgG H+L) (ab150165) at 1/2000 dilution for 30 min at 4ºC. Isotype control antibody (black line) was rat IgG2b [RTK4530] (ab18541, 1µg/1x10^6 cells) used under the same conditions. Unlabelled sample (blue line) was also used as a control. Acquisition of >5,000 events were collected using a 20mW Argon ion laser (488nm) and 525/30 bandpass filter.
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