Anti-CD68 antibody (ab125212)

False colour image of Western blot: Anti-CD68 antibody staining at 0.2 µg/ml, shown in green; Mouse anti-Alpha Tubulin [DM1A] (ab7291) loading control staining at 1/20000 dilution, shown in red. In Western blot, ab125212 was shown to bind specifically to CD68. A band was observed at 95-102 kDa in wild-type RAW 264.7 cell lysates with no signal observed at this size in CD68 knockout cell line ab280047 (knockout cell lysate ab280106). To generate this image, wild-type and CD68 knockout RAW 264.7 cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in fluorescent western blot (TBS-based) blocking solution before incubation with primary antibodies overnight at 4°C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit IgG H&L (IRDye^® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preabsorbed (ab216776) at 1/20000 dilution.

Paraffin-embedded rat liver tissue stained for CD68 using ab125212 at 1 μg/ml in immunohistochemical analysis.

ab125212 staining CD68 in Mouse spleen tissue sections by Immunohistochemistry (IHC-P - paraformaldehyde-fixed, paraffin-embedded sections). Tissue was fixed with paraformaldehyde and blocked with 5% serum for 30 minutes at 20°C; antigen retrieval was by heat mediation in a citrate buffer. Samples were incubated with primary antibody (1/100 in PBS + 2% BSA + 10% FCS) for 45 minutes at 20°C. A HRP-conjugated goat anti-rabbit IgG polyclonal (1/200) was used as the secondary antibody.

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CD68 was detected in paraffin-embedded section of mouse liver tissues. Heat mediated antigen retrieval was performed in citrate buffer (pH6, epitope retrieval solution ) for 20 mins. The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1μg/mL ab125212 overnight at 4°C. Cy3 Conjugated Goat Anti-Rabbit IgG was used as secondary antibody at 1:100 dilution and incubated for 30 minutes at 37°C. The section was counterstained with DAPI. Visualize using a fluorescence microscope and filter sets appropriate for the label used.

Western blot analysis of rat and mouse tissue lysates, labelling CD68 with ab125212

Paraffin-embedded mouse spleen tissue stained for CD68 using ab125212 at 1 μg/ml in immunohistochemical analysis.

Immunohistochemistry of microglial marker in mouse brain

Cortical staining for the microglial marker, CD68 increased as a function of age in rTg4510 animals, but remained unchanged in tTA animals. Scale bar, 200 µm, inset scale bar, 50 µm.

(After Figure 10 A of Wes et al.)

Frozen sectioned rat liver tissue stained for CD68 using ab125212 at 1 μg/ml in immunohistochemical analysis.

ab125212 staining CD68 in Mouse skin tissue sections by Immunohistochemistry (IHC-P - paraformaldehyde-fixed, paraffin-embedded sections). Tissue was fixed with formaldehyde and blocked with 10% serum for 30 minutes at 24°C; antigen retrieval was by heat mediation in a citrate buffer. Samples were incubated with primary antibody (1/2000 in 10% goat serum) for 16 hours at 4°C. A Biotin-conjugated goat anti-rabbit IgG polyclonal (1/500) was used as the secondary antibody.

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