CD68/SR-D1 Antibody (A3C4-R)

Western Blot: CD68/SR-D1 Antibody (A3C4-R) [NBP3-32165] - Western blot analysis of CD68/SR-D1 on different lysates with Mouse anti-CD68/SR-D1 antibody (NBP3-32165) at 1/1,000 dilution. Lane 1: A431 cell lysate Lane 2: THP-1 cell lysate Lane 3: U-937 cell lysate Lane 4: Jurkat cell lysate (negative) Lysates/proteins at 20 ug/Lane. Predicted band size: 37 kDa Observed band size: 100-150 kDa Exposure time: 3 minutes; 4-20% SDS-PAGE gel. Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (NBP3-32165) at 1/1,000 dilution was used in 5% NFDM/TBST at 4C overnight. Goat Anti-Mouse IgG - HRP Secondary Antibody at 1/50,000 dilution was used for 1 hour at room temperature.

Immunohistochemistry: CD68/SR-D1 Antibody (A3C4-R) [NBP3-32165] - Immunohistochemical analysis of paraffin-embedded human tonsil tissue with Mouse anti-CD68/SR-D1 antibody (NBP3-32165) at 1/1,000 dilution. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (NBP3-32165) at 1/1,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

Immunocytochemistry/ Immunofluorescence: CD68/SR-D1 Antibody (A3C4-R) [NBP3-32165] - Immunocytochemistry analysis of THP-1 (positive) and Jurkat (negative) labeling CD68/SR-D1 with Mouse anti-CD68/SR-D1/SR-D1 antibody (NBP3-32165) at 1/100 dilution. Cells were fixed in 100% precooled methanol for 5 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Mouse anti-CD68/SR-D1 antibody (NBP3-32165) at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Mouse IgG H&L (iFluor™ 488) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI. beta Tubulin (red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Rabbit IgG H&L (iFluor™ 594) were used as the secondary antibody at 1/1,000 dilution.

• Reactivity: Hu
• Applications: WB, ICC/IF, IHC
• Clone: A3C4-R
• Clonality: Monoclonal
• Host: Mouse
• Conjugate: Unconjugated
• Concentration: 1 mg/ml

CD68/SR-D1 Antibody (A3C4-R) Summary

• Additional Information: Recombinant Monoclonal Antibody
• Immunogen: Synthetic peptide within human CD68/SR-D1 aa 320-354. (Uniprot: P34810)
• Localization: Cell membrane. Endosome membrane, lysosome membrane.
• Isotype: IgG1
• Clonality: Monoclonal
• Host: Mouse
• Gene: CD68
• Purity: Protein A purified

Applications/Dilutions

• Dilutions:
  • Immunocytochemistry/ Immunofluorescence 1:100
  • Immunohistochemistry
  • Immunohistochemistry-Paraffin 1:1000
  • Western Blot 1:1000
• Theoretical MW: 37 kDa.
  Disclaimer note: The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors.

Packaging, Storage & Formulations

• Storage: Store at 4C short term. Aliquot and store at -20C long term. Avoid freeze-thaw cycles.
• Buffer: PBS (pH7.4), 0.1% BSA and 40% Glycerol
• Preservative: 0.05% Sodium Azide
• Concentration: 1 mg/ml
• Purity: Protein A purified

Alternate Names for CD68/SR-D1 Antibody (A3C4-R)

• 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

Human CD68, also known as GP110, LAMP4, Scavenger Receptor D1 (SR-D1) or macrosialin in mouse, encodes a 110-kD transmembrane glycoprotein that belongs to the lysosomal/endosomal-associated membrane glycoprotein (LAMP) family. Members of the LAMP family include LAMP-1, LAMP-2, dendritic cell (DC)-LAMP (aka CD208), and brain and dendritic cell-associated (BAD)-LAMP (aka LAMP-5). Unlike the two LAMP domains facing the lysosomal lumen in LAMP-1 and LAMP-2, CD68 has a single LAMP domain containing four cystines spaced 36-37 residues apart along with an N-terminal Mucin-like domain. The 354 amino acid (a.a.) human CD68 and 326 a.a. murine ortholog share 80.6% a.a. sequence identity (1).

CD68 is highly expressed in cells of the mononuclear phagocyte system such as macrophages, microglia, osteoclasts, and myeloid dendritic cells (DCs); and is expressed to a lesser extent in lymphoid cells (CD19+ B lymphocytes and CD4+ T lymphocytes), human umbilical cord mesenchymal stem cells (MSCs), fibroblasts, endothelial cells, multiple non-hematopoietic cancer cell lines, and human arterial intimal smooth muscle cells (SMCs). Expression has been also observed in diseased states for granulocytes and neutrophils, in particular basophils from myeloproliferative disorders and intestinal neutrophils from inflammatory bowel disease (IBD), respectively (1).

Although the function of CD68 has yet to be established, it has often been used as an immunohistochemistry (IHC) marker of inflammation and for granular cell tumors (GCTs). CD68+ tumor associated macrophages (TAMs) has been suggested to be a predictive marker for poor cancer prognosis, but a meta-analysis showed the presence of CD68 is not correlated with survival (2). In addition, a role in hepatic malaria infection has been reported based on the finding that peptide P39 binds CD68, considered a receptor for malaria sporozoite, and inhibits parasite entry into Kupffer cells. CD68 was deemed a member of the Scavenger receptor family due to its upregulation in macrophages following inflammatory stimuli, ability to bind modified LDL, phosphatidylserine, and apoptotic cells, as well as shuttling between the plasma membrane and endosomes. CD68 has been linked to atherogenesis based on binding and internalization of its ligand, oxLDL (1).

References

1. Chistiakov, DA, Killingsworth, MC, Myasoedova, VA. Orekhov AN, Bobryshev YV. (2017) CD68/macrosialin: not just a histochemical marker. Lab Invest. 97:4-13. PMID: 27869795

2. Troiano G, Caponio VCA, Adipietro I, Tepedino M, Santoro R, Laino L, Lo Russo L, Cirillo N, Lo Muzio L. (2019) Prognostic significance of CD68+ and CD163+ tumor associated macrophages in head and neck squamous cell carcinoma: A systematic review and meta-analysis. Oral Oncol. 93:66-75. PMID: 31109698.

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

• Gene Symbol: CD68