Anti-CD68 antibody is a rabbit polyclonal antibody that is used in CD68 western blot (WB) and immunohistochemistry (IHC). Suitable for mouse and rat samples.
-Cited in >720 publications.
- Affinity purified
Same trusted quality, new lower price
IgG
Rabbit
Constituents: 5.61% Trehalose, 0.45% Sodium chloride, 0.1% Disodium hydrogenorthophosphate
Liquid
Polyclonal
IHC-P | WB | IHC-Fr | |
---|---|---|---|
Mouse | Tested | Tested | Expected |
Rat | Tested | Tested | Tested |
Species | Dilution info | Notes |
---|---|---|
Species Mouse | Dilution info 0.5-1 µg/mL | Notes Perform heat-mediated antigen retrieval before commencing with IHC staining protocol. |
Species Rat | Dilution info 0.5-1 µg/mL | Notes Perform heat-mediated antigen retrieval before commencing with IHC staining protocol. |
Species | Dilution info | Notes |
---|---|---|
Species Mouse | Dilution info 0.1-0.5 µg/mL | Notes The detection limit for ab125212 is approximately 0.1ng/lane under non-reducing and reducing conditions. |
Species Rat | Dilution info 0.1-0.5 µg/mL | Notes The detection limit for ab125212 is approximately 0.1ng/lane under non-reducing and reducing conditions. |
Species | Dilution info | Notes |
---|---|---|
Species Rat | Dilution info 0.5-1 µg/mL | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Mouse | Dilution info Use at an assay dependent concentration. | Notes - |
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Could play a role in phagocytic activities of tissue macrophages, both in intracellular lysosomal metabolism and extracellular cell-cell and cell-pathogen interactions. Binds to tissue- and organ-specific lectins or selectins, allowing homing of macrophage subsets to particular sites. Rapid recirculation of CD68 from endosomes and lysosomes to the plasma membrane may allow macrophages to crawl over selectin-bearing substrates or other cells.
Macrosialin, Cd68
Anti-CD68 antibody is a rabbit polyclonal antibody that is used in CD68 western blot (WB) and immunohistochemistry (IHC). Suitable for mouse and rat samples.
-Cited in >720 publications.
- Affinity purified
Same trusted quality, new lower price
IgG
Rabbit
Constituents: 5.61% Trehalose, 0.45% Sodium chloride, 0.1% Disodium hydrogenorthophosphate
Liquid
Polyclonal
Affinity purification Immunogen
Blue Ice
+4°C
-20°C
Upon delivery aliquot
Avoid freeze / thaw cycle
For WB, as CD68 is highly glycosylated, it typically runs between 75-110 kDa depending on the amount of glycosylation in the sample.
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This supplementary information is collated from multiple sources and compiled automatically.
CD68 is a glycoprotein that functions as a scavenger receptor. It plays an important role in the regulation of cellular debris and apoptotic cell clearance. Researchers often refer to it as KP1 or macrosialin in the literature. CD68 with a molecular weight of approximately 110 kDa is highly expressed in macrophages and monocytes. The mucin-like structure of CD68 allows it to be heavily glycosylated which aids in its function. Researchers detect CD68 in tissues through methods like immunohistochemistry (IHC) immunofluorescence and CD68 staining and it is widely used as a macrophage marker in research.
CD68 facilitates functions associated with the innate immune response. It serves as an important part of macrophages which contribute to immune surveillance and tissue homeostasis. Though it primarily operates on its own in some contexts CD68 might assist in forming complexes with other receptors to modulate phagocytic activity. This target is significant in atherosclerotic plaque stability as macrophages engulf lipids and cell debris through processes facilitated by CD68.
CD68 is involved in pathways connected to inflammation and phagocytosis. The CD68 protein works closely with other scavenger receptors like CD36 to mediate uptake of oxidized low-density lipoproteins (oxLDL) in the lipid metabolism pathway. Additionally CD68 engagement can influence the toll-like receptor (TLR) signaling pathways thereby linking innate immunity and inflammatory responses critical for host defense and disease progression.
CD68 is associated with atherosclerosis and multiple sclerosis. In atherosclerosis its role becomes evident as it accumulates in macrophages within the plaques making it a marker of disease severity. In multiple sclerosis CD68 expression in macrophages and microglia indicates active demyelination and inflammation. It also relates to proteins such as CD36 in atherosclerosis where both mediate the uptake of modified LDL contributing to foam cell formation in plaques.
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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.)
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.
False colour image of Western blot: Anti-CD68 antibody staining at 0.2 μg/ml, shown in green; Mouse anti-Alpha Tubulin [DM1A] (Anti-alpha Tubulin antibody [DM1A] - Loading Control 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 Mouse CD68 knockout RAW 264.7 cell line ab280047 (knockout cell lysate Mouse CD68 knockout RAW 264.7 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 (Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed ab216776) at 1/20000 dilution.
All lanes: Western blot - Anti-CD68 antibody (ab125212) at 0.2 µg/mL
Lane 1: Wild-type RAW 264.7 cell lysate at 20 µg
Lane 2: CD68 knockout RAW 264.7 cell lysate at 20 µg
Lane 3: Mouse spleen cell lysate at 20 µg
Lane 4: Neuro-2a cell lysate at 20 µg
Performed under reducing conditions.
Predicted band size: 37 kDa
Observed band size: 95-102 kDa
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.
Immunohistochemical analysis of murine spleen tissue, staining CD68 with ab125212.
Tissue was fixed with paraformaldehyde and blocked with 5% serum for 1 hour at room temperature; antigen retrieval was by heat mediation in citrate buffer (pH 6). Samples were incubated with primary antibody (undiluted) for 16 hours at 4°C. An undiluted HRP-conjugated horse anti-rat polyclonal IgG was used as the secondary antibody.
Immunohistochemical analysis of paraffin-embedded Rat lung tissue labeling CD68 with ab125212 at 2 ?g/ml. Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with ab125212 overnight at 4°C. Peroxidase Conjugated Goat Anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using a HRP conjugated Rabbit IgG with DAB as the chromogen.
Immunohistochemical analysis of paraffin-embedded Rat spleen tissue labeling CD68 with ab125212 at 2 ?g/ml. Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with ab125212 overnight at 4°C. Peroxidase Conjugated Goat Anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using a HRP conjugated Rabbit IgG with DAB as the chromogen.
Immunohistochemical analysis of paraffin-embedded Mouse spleen tissue labeling CD68 with ab125212 at 2 ?g/ml. Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with ab125212 overnight at 4°C. Peroxidase Conjugated Goat Anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using a HRP conjugated Rabbit IgG with DAB as the chromogen.
Electrophoresis was performed on a 5-20% SDS-PAGE gel at 70V (Stacking gel) / 90V (Resolving gel) for 2-3 hours. The sample well of each lane was loaded with 30 ug of sample under reducing conditions. After electrophoresis, proteins were transferred to a nitrocellulose membrane at 150 mA for 50-90 minutes. Blocked the membrane with 5% non-fat milk/TBS for 1.5 hour at RT. The membrane was incubated with ab125212 at 0.5 ?g/mL overnight at 4°C, then washed with TBS-0.1%Tween 3 times with 5 minutes each and probed with a goat anti-rabbit IgG-HRP secondary antibody at 1/5000 dilution for 1.5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit with Tanon 5200 system. A specific band was detected for CD68 at approximately 90-100 kDa.
All lanes: Western blot - Anti-CD68 antibody (ab125212) at 0.5 µg/mL
Lane 1: Rat spleen tissue lysates
Lane 2: Mouse spleen tissue lysates
Lane 3: RAW264.7 whole cell lysates
All lanes: Goat anti-rabbit IgG-HRP at 1/5000 dilution
Immunohistochemical analysis of paraffin-embedded Mouse liver tissue labeling CD68 with ab125212 at 2 ?g/ml. Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with ab125212 overnight at 4°C. Peroxidase Conjugated Goat Anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using a HRP conjugated Rabbit IgG with DAB as the chromogen.
Immunohistochemical analysis of paraffin-embedded Rat liver tissue labeling CD68 with ab125212 at 2 ?g/ml. Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with ab125212 overnight at 4°C. Peroxidase Conjugated Goat Anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using a HRP conjugated Rabbit IgG with DAB as the chromogen.
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