Anti-CD68 antibody [FA-11] (ab53444)
Key features and details
- Rat monoclonal [FA-11] to CD68
- Suitable for: ICC, IHC-Fr, Flow Cyt (Intra)
- Reacts with: Mouse
- Isotype: IgG2a
Related conjugates and formulations
Overview
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Product name
Anti-CD68 antibody [FA-11]
See all CD68 primary antibodies -
Description
Rat monoclonal [FA-11] to CD68 -
Host species
Rat -
Tested applications
Suitable for: ICC, IHC-Fr, Flow Cyt (Intra)more details -
Species reactivity
Reacts with: Mouse -
Immunogen
Full length native protein (purified). This information is proprietary to Abcam and/or its suppliers.
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Positive control
- IHC-Fr: Mouse lung, spleen and heart tissue sections; Flow Cyt: RAW 264.7 cells
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General notes
Although some customers have had success with this antibody in IHC-P, we are unable to obtain positive results in this application and so cannot recommend it for IHC-P. We batch test the antibody in IHC-Fr.
This antibody clone is manufactured by Abcam. If you require a custom buffer formulation or conjugation for your experiments, please contact orders@abcam.com.
The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing this with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation. Please check that this product meets your needs before purchasing.
If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, along with publications, customer reviews and Q&As
Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. -
Storage buffer
pH: 7.40
Preservative: 0.02% Sodium azide
Constituents: PBS, 6.97% L-Arginine -
Concentration information loading...
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Purity
Protein G purified -
Purification notes
Purified IgG prepared by affinity chromatography on Protein G from tissue culture supernatant. -
Clonality
Monoclonal -
Clone number
FA-11 -
Isotype
IgG2a -
Research areas
Associated products
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Alternative Versions
- Alexa Fluor® 405 Anti-CD68 antibody [FA-11] (ab199571)
- Alexa Fluor® 488 Anti-CD68 antibody [FA-11] (ab201844)
- Alexa Fluor® 647 Anti-CD68 antibody [FA-11] (ab201845)
- HRP Anti-CD68 antibody [FA-11] (ab216526)
- PE Anti-CD68 antibody [FA-11] (ab216701)
- PerCP Anti-CD68 antibody [FA-11] (ab220509)
- APC Anti-CD68 antibody [FA-11] (ab221251)
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Compatible Secondaries
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Isotype control
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Recombinant Protein
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Related Products
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab53444 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Application | Abreviews | Notes |
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ICC |
Use a concentration of 1 µg/ml.
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IHC-Fr | (11) |
Use a concentration of 0.1 - 5 µg/ml.
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Flow Cyt (Intra) |
Use 0.2µg for 106 cells.
Membrane permeabilisation is required for this application. ab18450 - Rat monoclonal IgG2a, is suitable for use as an isotype control with this antibody. |
Notes |
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ICC
Use a concentration of 1 µg/ml. |
IHC-Fr
Use a concentration of 0.1 - 5 µg/ml. |
Flow Cyt (Intra)
Use 0.2µg for 106 cells. Membrane permeabilisation is required for this application. ab18450 - Rat monoclonal IgG2a, is suitable for use as an isotype control with this antibody. |
Target
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Function
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. -
Tissue specificity
Highly expressed by blood monocytes and tissue macrophages. Also expressed in lymphocytes, fibroblasts and endothelial cells. Expressed in many tumor cell lines which could allow them to attach to selectins on vascular endothelium, facilitating their dissemination to secondary sites. -
Sequence similarities
Belongs to the LAMP family. -
Post-translational
modificationsN- and O-glycosylated. -
Cellular localization
Cell membrane and Endosome membrane. Lysosome membrane. - Information by UniProt
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Database links
- Entrez Gene: 12514 Mouse
- SwissProt: P31996 Mouse
- Unigene: 15819 Mouse
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Alternative names
- CD 68 antibody
- CD68 antibody
- CD68 antigen antibody
see all
Images
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IHC image of CD68 staining in a section of frozen mouse lung performed on a Leica BOND™ system using the standard protocol. The section was fixed in 10% paraformaldehyde (10 min) prior to staining. The section was incubated with ab53444, 0.1ugml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX. The inset secondary-only control image is taken from an identical assay without primary antibody.
For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.
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ab53444 staining CD68 in Raw264.7 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% n
normal goat serum/0.3M glycine in 0.1%PBS-Tween for 1h. The cells were then incubated overnight at 4°C with ab53444 at 1µg/ml and ab6046, Rabbit polyclonal to beta Tubulin - Loading Control. Cells were then incubated with ab150165, Goat polyclonal Secondary Antibody to Rat IgG - H&L (Alexa Fluor® 488), pre-adsorbed 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.
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Immunohistochemistry (Frozen sections) - Anti-CD68 antibody [FA-11] (ab53444)Lee J. et al PLoS One. 2017 Nov 2;12(11):e0187189. doi: 10.1371/journal.pone.0187189. eCollection 2017. Reproduced under the Creative Commons license http://creativecommons.org/licenses/by/4.0/
Dual fluorescence combining IL-6 with markers for macrophages (CD68).
O, P, and Q, dual labeling of IL-6 (red) and marker of macrophage (green) in db/db mouse heart tissues. Arrows in Q show the specific CD68 staining with absence of IL-6 staining. (R and S) Negative control: arrows show the absence of staining in vessels with control IgG and without primary antibodies. (T) Staining of nuclei with DAPI (blue) in heart tissues of the db/db mice.
To identify and localize IL-6 protein in coronary arterioles, transverse sections of the mouse heart were stained using markers of endothelial cells, vascular smooth muscle cells, and macrophages. Freshly isolated hearts were embedded and frozen in OCT and sectioned at 5 μm. Slides were incubated with blocking solution (10% donkey serum in PBS) and permeabilized (0.1% Triton X-100 in PBS). Primary antibodies to IL-6 (goat polyclonal 15 micro g/ml, AF-406-NA; R&D) or macrophage marker CD68 (rat monoclonal, 1:1000, ab53444; Abcam) were used for sequential double immunofluorescence staining. Secondary antibodies were conjugated with the fluorophores FITC or Texas red. Sections were mounted in an anti-fading agent (Slowfade gold with DAPI; Invitrogen), and then the slides were observed and analyzed with a fluorescence microscope (IX81; Olympus) with a x40 objective (0.90 numerical aperture). For negative controls, primary antibodies were replaced with goat polyclonal IgG (Abcam), rabbit polyclonal IgG (GeneTex), and rat monoclonal IgG (Abcam) isotype controls at the same concentration. The specificity of the primary antibody was confirmed as the absence of immunofluorescence staining signals in the IL-6-/- mice.
Diabetic mice (db/db).
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ab53444 staining CD68 in mouse spleen tissue by Immunohistochemistry (Frozen sections). Antibody was detected with HRP-conjugated Goat anti-Rat IgG, showing staining in the red pulp.
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Immunohistochemistry (Frozen sections) - Anti-CD68 antibody [FA-11] (ab53444)This image is courtesy of an Abreview submitted by Miss Silke Vorwald
ab53444 staining mouse heart tissue by Immunohistochemistry (IHC-frozen sections). Tissue underwent fixation in paraformaldehyde, no permeabilization and blocked in 1% serum for 10 minutes at 20°C. The primary antibody used undiluted and incubated with sample for 16 hours at 20°C. A Biotin conjugated goat polyclonal to rat Ig, diluted 1/100 was used as the secondary.
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Immunohistochemistry (Frozen sections) - Anti-CD68 antibody [FA-11] (ab53444)This image is courtesy of an anonymous Abreviewab53444 at 1/100 dilution staining CD68 in mouse spleen tissue by immunohistochemistry (frozen sections). Sections were acetone fixed prior to blocking in 8% milk for 40 minutes at 36°C and then incubated with ab53444 for 20 hours at 4°C. A biotin conjugated goat polyclonal to rat Ig, diluted 1/400, was used as the secondary antibody.
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Immunohistochemistry (Frozen sections) - Anti-CD68 antibody [FA-11] (ab53444)This image is courtesy of an anonymous Abreview.ab53444 staining CD68 in murine heart tissue by Immunohistochemistry (Frozen sections).
Tissue was fixed in acetone, permeabilized using 0.3% Triton, blocked with 10% serum for 30 minutes at 20°C, then incubated with ab53444 at a 1/1000 dilution for 16 hours at 4°C. The secondary used was a FITC conjugated goat anti-rat polyclonal, used at a 1/1000 dilution. -
Flow cytometry overlay histogram showing RAW 264.7 cells stained with ab53444 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Triton X-100 for 15 min. The cells were then incubated in 1x PBS containing 10% normal goat serum to block non-specific protein-protein interaction followed by the antibody (ab53444) (1x106 in 100µl at 0.2µg/ml) for 30 min at 22°C. The secondary antibody Goat anti-mouse IgG H&L (Alexa Fluor 488®, pre-adsorbed) (ab150117) was used at 1/2000 for 30 min at 22°C.
Isotype control antibody (black line) was Rat IgG2a k (ab18450) used at the same concentration and conditions as the primary antibody. Unlabelled sample (blue line) was also used as a control.
Acquisition of >5000 events were collected using a 50 mW Blue laser (488nm) and 525/40 bandpass filter.
This antibody gave a positive signal in RAW 264.7 cells fixed with 4% formaldehyde (10 min) / permeabilized with 0.1% PBS-Triton X-100 for 15 min used under the same conditions.
Protocols
Datasheets and documents
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SDS download
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Datasheet download
References (205)
ab53444 has been referenced in 205 publications.
- Pendharkar AV et al. Optogenetic Stimulation Reduces Neuronal Nitric Oxide Synthase Expression After Stroke. Transl Stroke Res 12:347-356 (2021). PubMed: 32661768
- Yu Q et al. Dietary Cholesterol Exacerbates Statin-Induced Hepatic Toxicity in Syrian Golden Hamsters and in Patients in an Observational Cohort Study. Cardiovasc Drugs Ther 35:367-380 (2021). PubMed: 32860619
- Gula G et al. Potential functions of embryonic cardiac macrophages in angiogenesis, lymphangiogenesis and extracellular matrix remodeling. Histochem Cell Biol 155:117-132 (2021). PubMed: 33130914
- Klemke L et al. Hsp90-stabilized MIF supports tumor progression via macrophage recruitment and angiogenesis in colorectal cancer. Cell Death Dis 12:155 (2021). PubMed: 33542244
- Flores-Pliego A et al. Molecular Insights into the Thrombotic and Microvascular Injury in Placental Endothelium of Women with Mild or Severe COVID-19. Cells 10:N/A (2021). PubMed: 33578631