CD163 Antibody (1113338) [Unconjugated]

CD163 was detected in immersion fixed paraffin-embedded sections of mouse spleen using Rat Anti-Mouse CD163, Monoclonal Antibody (Catalog # MAB11745) at 3 μg/mL at 37° Celsius for 4 minutes. Before incubation with the primary antibody, tissue underwent an all-in-one dewaxing and antigen retrieval preprocessing using PreTreatment Module (PT Module) and Dewax and HIER Buffer H (pH 9; Epredia Catalog # TA-999-DHBH).Tissue was stained using the Alexa Fluor™ 647 Goat anti-Rat IgG Secondary Antibody at 1:200 at 37 ° Celsius for 2 minutes. (Yellow; Lunaphore Catalog # DR647RT) and counterstained with DAPI (blue; Lunaphore Catalog # DR100). Specific staining was localized to the cytoplasm. Protocol available in COMET™ Panel Builder.

Western Blot shows lysates of mouse spleen tissue. PVDF membrane was probed with 2 µg/ml of Rat Anti-Mouse CD163 Monoclonal Antibody (Catalog # MAB11745) followed by HRP-conjugated Anti-Rat IgG Secondary Antibody (Catalog # HAF005). A specific band was detected for CD163 at approximately 160 kDa (as indicated). This experiment was conducted under reducing conditions and using Western Blot Buffer Group 1.

CD163 was detected in perfusion fixed paraffin-embedded sections of mouse colon using Rat Anti-Mouse CD163 Monoclonal Antibody (Catalog # MAB11745) at 5 µg/ml overnight at 4 °C. Before incubation with the primary antibody, tissue was subjected to heat-induced epitope retrieval using VisUCyte Antigen Retrieval Reagent-Basic (Catalog # VCTS021). Tissue was stained using the HRP-conjugated Anti-Rat IgG Secondary Antibody (Catalog # HAF005) and counterstained with hematoxylin (blue). Specific staining was localized to the cell surface. View our protocol for Chromogenic IHC Staining of Paraffin-embedded Tissue Sections.

CD163 was detected in perfusion fixed paraffin-embedded sections of mouse liver using Rat Anti-Mouse CD163 Monoclonal Antibody (Catalog # MAB11745) at 5 µg/ml overnight at 4 °C. Before incubation with the primary antibody, tissue was subjected to heat-induced epitope retrieval using VisUCyte Antigen Retrieval Reagent-Basic (Catalog # VCTS021). Tissue was stained using the HRP-conjugated Anti-Rat IgG Secondary Antibody (Catalog # HAF005) and counterstained with hematoxylin (blue). Specific staining was localized to the cell surface of Kupffer cells. View our protocol for Chromogenic IHC Staining of Paraffin-embedded Tissue Sections.

• Reactivity: Mu
• Applications: WB, IHC, mIF
• Clone: 1113338
• Clonality: Monoclonal
• Host: Rat
• Conjugate: Unconjugated

CD163 Antibody (1113338) [Unconjugated] Summary

• Immunogen: Synthetic Peptide
  Accession # Q2VLH6
• Specificity: Detects a synthetic peptide specific for mouse CD163 around amino acid 920 in Direct ELISA.
• Source: N/A
• Isotype: IgG2b
• Clonality: Monoclonal
• Host: Rat
• Purity Statement: Protein A or G purified from hybridoma culture supernatant

Applications/Dilutions

• Dilutions:
  • Immunohistochemistry 3-25 ug/mL
  • Multiplex Immunofluorescence 3 ug/mL
  • Western Blot 2 ug/mL

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.
• Reconstitution Instructions: Reconstitute lyophilized material at 0.2 mg/ml in sterile PBS. For liquid material, refer to CoA for concentration.

Notes

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

Alternate Names for CD163 Antibody (1113338) [Unconjugated]

• CD_antigen: CD163
• CD163 molecule
• CD163
• GHI/61
• HbSR
• Hemoglobin scavenger receptor
• M130
• macrophage-associated antigen
• MM130
• RM3/1
• SCARI1
• scavenger receptor cysteine-rich type 1 protein M130
• sCD163
• Soluble CD163

Background

CD163, also known as M130 and  p155, is a 130-160 kDa type I transmembrane protein belonging to group B of the cysteine-rich scavenger receptor family (1). Mature mouse CD163 consists of a 1007 amino acid (aa) extracellular domain (ECD) with 9 scavenger receptor cysteine-rich (SRCR) domains, a 21 aa transmembrane segment, and a 55 aa cytoplasmic domain (2). Alternative splicing of mouse CD163 generates an isoform with a substitution in the cytoplasmic region. Within the ECD, mouse CD163 shares 75% and 90% aa sequence identity with human and rat CD163, respectively. CD163 is expressed on monocytes and macrophages and is inducible by immunosuppressant glucocorticoids and IL-10 (3-5). A soluble form is shed from the cell surface by TACE or neutrophil elastase mediated cleavage (6, 7) in response to oxidative stress, Prostaglandin F2a stimulation, or the activation of Fc gamma receptors, TLR1, 2, 5, or 6 (8-10). CD163 mediates monocyte binding to bacteria, leading to the release of inflammatory cytokines (11). It is essential for the circulatory clearance of hemoglobin-haptoglobin (Hb-Hp) complexes as well as free hemoglobin (12, 13). It can also mediate monocyte-erythroblast adhesion and promote erythroblast expansion (14). CD163 binds and internalizes the cytokine TWEAK, and the ratio of soluble CD163 to TWEAK in the plasma is elevated during atherosclerosis (15, 16).

1. Kowal, K. et al. (2011) Folia Histochem. Cytobiol. 49:365.
2. Law, S.K.A. et al. (1993) Eur. J. Immunol. 23:2320.
3. Hogger, P. et al. (1998) J. Immunol. 161:1883.
4. Sulahian, T.H. et al. (2000) Cytokine 12:1312.
5. Buechler, C. et al. (2000) J. Leukoc. Biol. 67:97.
6. Etzerodt, A. et al. (2010) J. Leukoc. Biol. 88:1201.
7. Moreno, J.A. et al. (2012) Eur. Heart J. 33:252.
8. Weaver, L.K. et al. (2006) J. Leukoc. Biol. 80:26.
9. Timmermann, M. and P. Hogger (2005) Free Radic. Biol. Med. 39:98.
10. Sulahian, T.H. et al. (2004) J. Leukoc. Biol. 76:271.
11. Fabriek, B.O. et al. (2009) Blood 113:887.
12. Kristiansen, M. et al. (2001) Nature 409:198.
13. Schaer, D.J. et al. (2006) Blood 107:373.
14. Fabriek, B.O. et al. (2007) Blood 109:5223.
15. Bover, L.C. et al. (2007) J. Immunol. 178:8183.
16. Moreno, J.A. et al. (2009) Atherosclerosis 207:103.

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.