CD68/SR-D1 Antibody (2449C) [Unconjugated]

CD68/SR-D1 was detected in immersion fixed RAW 264.7 mouse monocyte/macrophage cell line (positive stain) and NIH-3T3 mouse embryonic fibroblast cell line (negative stain) using Rabbit Anti-Mouse CD68/SR-D1 Monoclonal Antibody (Catalog # MAB10114) at 3 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Rabbit IgG Secondary Antibody (red; Catalog # NL004) and counterstained with DAPI (blue). Specific staining was localized to cytoplasm. View our protocol for Fluorescent ICC Staining of Non-adherent Cells.

Raw264.7 mouse cell line was stained with Rabbit Anti-Mouse CD68 Monoclonal Antibody (Catalog # MAB10114, filled histogram) or Normal Rabbit IgG Control (Catalog # MAB1050, open histogram) followed by Phycoerythrin-conjugated Anti-Rabbit IgG Secondary Antibody (Catalog # F0110). To facilitate intracellular staining, cells were fixed with Flow Cytometry Fixation Buffer (1x) (Catalog # FC004) and permeabilized with ice-cold methanol. View our protocol for Staining Intracellular Molecules.

Mechanical compression stimulated piezo‐type mechanosensitive ion channel component 1 (PIEZO1) signaling for CD68+F4/80+ macrophages differentiation in whole periosteal cells (WPCs) and bone marrow‐derived macrophages (BMDMs). A–I) Primary WPCs or BMDMs were harvested from WT mice and subjected to 10 or 0 kPa of mechanical compression. A) mRNA expression of Cd68 and adhesion G protein‐coupled receptor E1 (Adgre1) by RT‐qPCR in WPCs after compression. B) mRNA expression of Adgre1 in BMDMs after compression. C) Immunofluorescent staining of CD68+ (green) and F4/80+ (red) cells in BMDMs after compression. Blue indicates DAPI staining of nuclei. Scale bar, 5 µm. D,E) mRNA expression of colony‐stimulating factor 1 (Csf1) in WPCs and BMDMs after compression, respectively. F,G) mRNA expression of Piezo1 in WPCs and BMDMs after compression, respectively. H,I) BMDMs were pretreated with the PIEZO1 inhibitor GsMTx4 or vehicle for 24 h before compression. H) mRNA expression of Csf1 and Adgre1 in BMDMs after compression. I) Immunofluorescent staining of CD68+ (green) and F4/80+ (red) cells after compression. Blue indicates DAPI staining of nuclei. Scale bar, 5 µm. Data are presented as mean ± SEM. n = 3 per group. A,B,D–G) * p < 0.05 compared with 0 kPa. Statistical significance was determined by unpaired, two‐tailed Student's t‐test. H) *p < 0.05 compared with vehicle 10‐kPa compression group; †p < 0.05. Statistical significance was determined by two‐way ANOVA with Bonferroni post hoc test. Image collected and cropped by CiteAb from the following open publication (//pubmed.ncbi.nlm.nih.gov/34854257), licensed under a CC-BY license. Not internally tested by R&D Systems.

• Reactivity: Mu
• Applications: CyTOF-ready, Flow, ICC/IF
• Clone: 2449C
• Clonality: Monoclonal
• Host: Rabbit
• Conjugate: Unconjugated

CD68/SR-D1 Antibody (2449C) [Unconjugated] Summary

• Additional Information: Recombinant Monoclonal Antibody.
• Immunogen: Chinese Hamster Ovary cell line, CHO-derived mouse CD68
  Asp21-Pro282
  Accession # P31996
• Specificity: Detects mouse CD68 in direct ELISAs.
• Source: N/A
• Isotype: IgG
• Clonality: Monoclonal
• Host: Rabbit
• Purity Statement: Protein A or G purified from cell culture supernatant

Applications/Dilutions

• Dilutions:
  • CyTOF-ready
  • Immunocytochemistry 3-25 ug/mL
  • Intracellular Staining by Flow Cytometry 0.25 ug/10^6 cells

Packaging, Storage & Formulations

• Storage: Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
  • 12 months from date of receipt, -20 to -70 °C as supplied.
  • 1 month, 2 to 8 °C under sterile conditions after reconstitution.
  • 6 months, -20 to -70 °C under sterile conditions after reconstitution.
• Buffer: Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied either lyophilized or as a 0.2 µm filtered solution in PBS.
• Reconstitution Instructions: Reconstitute at 0.5 mg/mL in sterile PBS.

Notes

This product is produced by and ships from R&D Systems, Inc., a Bio-Techne brand.

Alternate Names for CD68/SR-D1 Antibody (2449C) [Unconjugated]

• CD_antigen: CD68
• CD68 antigen
• CD68 molecule
• CD68
• DKFZp686M18236
• gp110
• LAMP4
• macrophage antigen CD68
• Macrosialin
• SCARD1
• scavenger receptor class D, member 1
• SRD1
• SR-D1

Background

Macrosialin (CD68) is a macrophage-restricted glycosylated transmembrane protein.  It belongs to the lamp family and carries out specialized functions in dedicated phagocytic cells.  Macrosialin binds to tissue- and organ-specific lectins to allow homing of macrophages to specific targets.  Its presence in macrophages is useful in diagnosing conditions related to proliferation or abnormality of these cells.

Limitations

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

Bioinformatics

• Uniprot:
  • P31996()