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PMID: 7868861. The antigen is temperature sensitive; we may recommend following the conditions as recommended in this publication. As per publication low temperature paraffin, using 0.5 to 2% paraformaldehyde fixation instead of 4% or using periodate-lysine-paraformaldehyde (PLP) + 0.25% PFA and 0.1% glutaraldehyde might be helpful in detection. In case of any question please do not hesitate to contact technical team at firstname.lastname@example.org.
The principal use of this antibody is in the study of functional heterogeneity of macrophages; it may be used to follow macrophage differentiation and to investigate the function of the CD11b/c equivalent antigen.
Our Abpromise guarantee covers the use of ab1211 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|IHC-FoFr||Use at an assay dependent concentration.
The use of ABC will enable further dilution of the antibody, see Abreview.
|ELISA||1/100 - 1/3200.|
|Flow Cyt||1/25 - 1/200.
(10 µl per million cells).
|IHC-FrFl||Use at an assay dependent concentration.|
|ICC/IF||Use a concentration of 5 - 10 µg/ml.|
|IP||Use at an assay dependent concentration. PubMed: 3512425|
ab1211 staining CD11b/c in rat spleen tissue sections by Immunohistochemistry (PFA perfusion fixed frozen sections). Tissue samples were fixed with formaldehyde and blocked with BSA for 10 minutes at 21°C. The sample was incubated with primary antibody (1/2000 in TBS/BSA/azide) at 21°C for 16 hours. A Biotin-conjugated Goat anti mouse polyclonal (1/300) was used as the secondary antibody.
ab1211 stained NR8383 cells. The cells were 100% methanol fixed for 10 minutes and then incubated in 1%BSA / 10% normal Goat serum / 0.3M glycine in 0.1% PBS-Tween for 1hour at room temperature to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab1211 at 5µg/ml) overnight at +4°C. The secondary antibody (pseudo-colored green) was ab150117 Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) preadsorbed used at a 1/1000 dilution for 1hour at room temperature. Alexa Fluor® 594 WGA was used to label plasma membranes (pseudo-colored red) at a 1/200 dilution for 1hour 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.
ab1211 staining CD11 in Rat brain tissue sections by Immunohistochemistry (PFA perfusion fixed frozen sections). Tissue samples were fixed by perfusion with paraformaldehyde and blocked with NGS in 0.3% PBS-TritonX-100 for 60 minutes at 24°C. The sample was incubated with primary antibody (1/300 in NGS in 0.3% PBS-TritonX-100) at 4°C for 42 hours. Ab150111 (1/500) was used as the secondary antibody.
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