Anti-CD11b + CD11c antibody [OX42] (ab1211)

Immunofluorescence staining of CD11b + CD11c staining in a section of 10% formalin fixed (10 mins, RT) frozen rat spleen.

Non-specific protein-protein interactions were then blocked in TBS containing 0.025% (v/v) Triton X-100, 0.3M (w/v) glycine and 1% (w/v) BSA for 1h at room temperature. The section was then incubated overnight at +4°C in TBS containing 0.025% (v/v) Triton X-100 and 1% (w/v) BSA with ab1211 at 1µg/ml. The section was then incubated with ab150117 (Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) preabsorbed, (Shown in green) 1.5µg/ml) for 1 hour at room temperature. Nuclear DNA was labelled with DAPI (shown in blue). The section was then mounted using Fluoromount®.

Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

For other IHC staining systems (automated and non-automated), customers should optimize variable parameters such as antigen retrieval conditions, antibody concentrations and incubation times.

ab1211 staining CD11b + CD11c (shown in red) in rat spleen tissue sections by IHC (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 staining CD11b + CD11c in NR8383 cells. The cells were fixed with 4% paraformaldehyde (10 min), permeabilized with 0.1% PBS-Tween for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h. The cells were then incubated overnight at 4°C with ab1211 at 1 µg/ml and ab6046, Rabbit polyclonal to beta Tubulin - Loading Control. Cells were then incubated with ab150117, Goat polyclonal Secondary Antibody to Mouse IgG H&L (Alexa Fluor^® 488) preadsorbed at 1/1000 dilution (shown in green) and ab150080, Goat polyclonal Secondary Antibody to Rabbit IgG - H&L (Alexa Fluor^® 594) at 1/1000 dilution (shown in pseudocolour red). Nuclear DNA was labelled with DAPI (shown in blue).

Also suitable in cells fixed with 100% methanol (5 min).

Image was acquired with a high-content analyser (Operetta CLS, Perkin Elmer) and a maximum intensity projection of confocal sections is shown.

ab1211 staining CD11b + CD11c (shown in green) in rat spleen tissue by IHC (Frozen sections). Tissue was fixed in formaldehyde, blocked with 10% serum for 30 minutes at 24°C, then incubated with ab1211 at a 1/100 dilution. The secondary used was an Alexa-Fluor 488 conjugated goat anti-mouse polyclonal, used at a 1/1000 dilution.

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