CD68 KO cell line available to order. KO validated by Western blot. Free of charge wild type control provided.
Images
Key facts
- Cell type
- RAW 264.7
- Species or organism
- Mouse
- Tissue
- Lymphatic
- Form
- Liquid
- Knockout validation
- Sanger Sequencing, Western blot
Alternative names
CD68 antigen, CD68 molecule, CD68_HUMAN, DKFZp686M18236, GP11, Gp110, LAMP4, MACROPHAGE ANTIGEN CD68, Macrophage antigen CD68 (microsialin), Macrosialin, SCARD1, Scavenger receptor class D member 1
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CD68 KO cell line available to order. KO validated by Western blot. Free of charge wild type control provided.
Key facts
- Cell type
- RAW 264.7
- Species or organism
- Mouse
- Tissue
- Lymphatic
- Form
- Liquid
- Knockout validation
- Sanger Sequencing, Western blot
- Disease
- Carcinoma
- Concentration
Properties
- Gene name
- CD68
- Gene editing type
- Knockout
- Gene editing method
- CRISPR technology
- Knockout validation
- Sanger Sequencing, Western blot
Cell culture
- Biosafety level
- EU: 2 US: 2
- Adherent/suspension
- Adherent
- Gender
- Male
Handling procedures
- Initial handling guidelines
Upon arrival, the vial should be stored in liquid nitrogen vapor phase and not at -80°C. Storage at -80°C may result in loss of viability.
1. Thaw the vial in 37°C water for bath approximately 1-2 minutes.
2. Transfer the cell suspension (0.8 mL) to a 15 mL/50 mL conical sterile polypropylene centrifuge tube containing 8.4 mL pre-warmed culture medium, wash vial with an additional 0.8 mL culture medium (total volume 10 mL) to collect remaining cells, and centrifuge at 201 x g (rcf) for 5 minutes at room temperature. 10 mL represents minimum recommended dilution. 20 mL represents maximum recommended dilution.
3. Resuspend the cell pellet in 5 mL pre-warmed culture medium and count using a haemocytometer or alternative cell counting method all remaining cells into an ultra-low attachment T25.
4. Incubate the culture at 37°C incubator with 5% CO2. Check the culture one day after revival and continue to check until 80% confluent.
5. Once confluent passage into an appropriate flask at a density of 2x105 cells/cm2. Seeding density is given as a guide only and should be scaled to align with individual lab schedules. Cultures should be monitored daily.- Subculture guidelines
- Grow in ultra-low attachment flasks.
- Do not wash or use dissociation reagent. Cells are semi-adherent. Tap flask sharply to remove adhered cells.
- All seeding densities should be based on cell counts gained by established methods.
- A guide seeding density of 2x105 cells/mL is recommended.
- Cells should be passaged when they have achieved 80-90% confluence.
- Culture medium
- DMEM + 10% FBS
- Cryopreservation medium
- Cell Freezing Medium-DMSO Serum free media, contains 8.7% DMSO in MEM supplemented with methyl cellulose.
Storage
- Shipped at conditions
- Dry Ice
- Appropriate short-term storage conditions
- -196°C
- Appropriate long-term storage conditions
- -196°C
Notes
We will provide viable cells that proliferate on revival.
This product is subject to limited use licenses from The Broad Institute and ERS Genomics Limited, and is developed with patented technology. For full details of the limited use licenses and relevant patents please refer to our limited use license and patent pages.
Supplementary info
- This supplementary information is collated from multiple sources and compiled automatically.
- Activity summary
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.
- Biological function summary
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.
- Pathways
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.
- Associated diseases and disorders
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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3 product images
Western blot - Mouse CD68 knockout RAW 264.7 cell line (ab280047)
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 Anti-CD68 antibody 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 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°. 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 (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 2: Western blot - Mouse CD68 knockout RAW 264.7 cell line (ab280047)
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
Western blot - Mouse CD68 knockout RAW 264.7 cell line (ab280047)
Western blot: Anti-CD68 antibody [RM1031] (Anti-CD68 antibody [RM1031] ab303565) staining at 1/1000 dilution, shown in green; Mouse anti-GAPDH antibody [6C5] (Anti-GAPDH antibody [6C5] - Loading Control ab8245) loading control staining at 1/20000 dilution, shown in magenta. In Western blot, Anti-CD68 antibody [RM1031] ab303565 was shown to bind specifically to CD68. A band was observed at 100 kDa in wild-type RAW 264.7 cell lysates with no signal observed at this size in CD68 knockout cell line. 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 5% milk in TBS-0.1% Tween® 20 (TBS-T) 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 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution.
All lanes: Western blot - Anti-CD68 antibody [RM1031] (Anti-CD68 antibody [RM1031] ab303565)
Lane 1: Wild-type RAW 264.7 cell lysate at 20 µg
Lane 2: CD68 knockout RAW 264.7 cell lysate at 20 µg
Developed using the ECL technique.
Performed under reducing conditions.
Observed band size: 100 kDa
Sanger Sequencing - Mouse CD68 knockout RAW 264.7 cell line (ab280047)
Mouse Cd68 KO in Raw264.7 Cells with 28 bp insertion-13 bp deletion-135 bp insertion-72 bp deletion in Exon 2.
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