Clone:
GHI/61.1
Type of antibody:
Primary antibodies
Isotype:
mouse IgG1κ
Applications:
FC, MICS, IF, IHC
Alternative names:
GHI/61, M130, MM130, CD163

Extended validation for CD163 Antibody, anti-human

Specificity

Epitope competition
In order to compare the epitope specificity of an antibody, the clone being used is compared with other known clones recognizing the same antigen in a competition assay.
Other clonesOverlap in epitope recognition with GHI/61.1
GHI/61++
215927-
REA812++
Cells were incubated with an excess of purified unconjugated CD163 (GHI/61.1) antibody followed by staining with fluorochrome-conjugated antibodies of other known clones against the same marker. Based on the fluorescence signal obtained, the clones were identified as recognizing completely overlapping (++), partially overlapping (+), or completely different epitopes (-) of the marker.

Sensitivity

Performance comparison
Selected fluorochrome conjugated antibodies from Miltenyi Biotec were compared to commercially available hybridoma clones in flow cytometry analysis.
View details
Flow cytometric comparison of different clones for CD163. Human peripheral blood mononuclear cells (PBMCs) were stained with CD163 antibodies and with a suitable counterstaining. As a control, CD163 antibody staining was omitted and cells were measured in the same channels. Flow cytometry was performed with the MACSQuant® Analyzer. Cell debris, dead cells, and cell doublets were excluded from the analysis based on scatter signals and 4',6-diamidino-2-phenylindole (DAPI) fluorescence. No FcR Blocking Reagent was used. The recommended titers of respective antibodies from different suppliers were used.
View details
Flow cytometric comparison of different clones for CD163. Human peripheral blood mononuclear cells (PBMCs) were stained with CD163 antibodies and with a suitable counterstaining. As a control, CD163 antibody staining was omitted and cells were measured in the same channels. Flow cytometry was performed with the MACSQuant® Analyzer. Cell debris, dead cells, and cell doublets were excluded from the analysis based on scatter signals and 4',6-diamidino-2-phenylindole (DAPI) fluorescence. No FcR Blocking Reagent was used. The recommended titers of respective antibodies from different suppliers were used.
View details
Flow cytometric comparison of different clones for CD163. Human peripheral blood mononuclear cells (PBMCs) were stained with CD163 antibodies and with a suitable counterstaining. As a control, CD163 antibody staining was omitted and cells were measured in the same channels. Flow cytometry was performed with the MACSQuant® Analyzer. Cell debris, dead cells, and cell doublets were excluded from the analysis based on scatter signals and 4',6-diamidino-2-phenylindole (DAPI) fluorescence. No FcR Blocking Reagent was used. The recommended titers of respective antibodies from different suppliers were used.
View details
Flow cytometric comparison of different clones for CD163. Human peripheral blood mononuclear cells (PBMCs) were stained with CD163 antibodies and with a suitable counterstaining. As a control, CD163 antibody staining was omitted and cells were measured in the same channels. Flow cytometry was performed with the MACSQuant® Analyzer. Cell debris, dead cells, and cell doublets were excluded from the analysis based on scatter signals and 4',6-diamidino-2-phenylindole (DAPI) fluorescence. No FcR Blocking Reagent was used. The recommended titers of respective antibodies from different suppliers were used.
Flow cytometric comparison of different clones for CD163. Human peripheral blood mononuclear cells (PBMCs) were stained with CD163 antibodies and with a suitable counterstaining. As a control, CD163 antibody staining was omitted and cells were measured in the same channels. Flow cytometry was performed with the MACSQuant® Analyzer. Cell debris, dead cells, and cell doublets were excluded from the analysis based on scatter signals and 4',6-diamidino-2-phenylindole (DAPI) fluorescence. No FcR Blocking Reagent was used. The recommended titers of respective antibodies from different suppliers were used.
Fixation data
To provide an indication on how an antibody performs after fixation of cells, in-house data on staining results before and after fixation with 3.7% formaldehyde using Miltenyi Biotec antibodies are provided. Different experimental settings may lead to different results.
No fixation
Post fixation
View details
Comparison of staining pattern on non-fixed and fixed cells using CD163 (GHI/61.1). The performance of the antibody after fixation was tested by comparing the staining pattern on fresh (no fixation) versus fixed (post fixation) cells.
View details
Comparison of staining pattern on non-fixed and fixed cells using CD163 (GHI/61.1). The performance of the antibody after fixation was tested by comparing the staining pattern on fresh (no fixation) versus fixed (post fixation) cells.
Comparison of staining pattern on non-fixed and fixed cells using CD163 (GHI/61.1). The performance of the antibody after fixation was tested by comparing the staining pattern on fresh (no fixation) versus fixed (post fixation) cells.

Specifications for CD163 Antibody, anti-human

Overview

The monoclonal antibody GHI/61.1 reacts with human CD163, a single-chain transmembrane protein also known as hemoglobin scavenger receptor or M130. It is expressed by mature tissue macrophages and peripheral blood monocytes. The expression of CD163 is up-regulated
in vitro
and
in vivo
by anti-inflammatory mediators such as IL-10 and (gluco)corticosteroid and is shed to blood upon inflammatory activation of macrophages. CD163 functions as a high affinity scavenger receptor for the complex of haemoglobin and haptoglobin. Depending on the ligand, crosslinking of M130 initiates signal transduction leading to the production of proinflammatory cytokines Il-1ß, IL-6, and GM-CSF or the antiinflammatory cytokine IL-10.

Alternative names

GHI/61, M130, MM130, CD163

Detailed product information

Technical specifications

CloneGHI/61.1
Clonalitymonoclonal
Isotypemouse IgG1κ
Isotype controlIsotype Control Antibody, mouse IgG1
Hostmouse
Type of antibodyPrimary antibodies
Specieshuman, non-human primate
Cross-reactivity
cynomolgus monkey (
Macaca fascicularis
)
AntigenCD163
Alternative names of antigenGHI/61, M130, MM130, CD163
Molecular mass of antigen [kDa]121
Distribution of antigenmacrophages, monocytes, CNS cells, thymocytes, bone marrow, liver, spleen
Entrez Gene ID9332
RRIDAB_2819455, AB_2655492, AB_2655493, AB_2655495, AB_2655496, AB_2655497, AB_2655498, AB_2655499, AB_2655500, AB_2655501, AB_2819470

References for CD163 Antibody, anti-human

Publications

  1. Kristiansen, M. et al. (2001) Identification of the haemoglobin scavenger receptor. Nature 409(6817): 198-201
  2. Law, S. K. et al. (1993) A new macrophage differentiation antigen which is a member of the scavenger receptor superfamily. Eur. J. Immunol. 23(9): 2320-2325
  3. Ritter, M. et al. (2001) Interaction of CD163 with the regulatory subunit of casein kinase II (CKII) and dependence of CD163 signaling on CKII and protein kinase C. Eur. J. Immunol. 31(4): 999-1009
  4. Pulford, K. et al. (1992) A monocyte/macrophage antigen recognized by the four antibodies GHI/61, Ber-MAC3, Ki-M8 and SM4. Immunology 75(4): 588-595
  5. Burdo, T. H. et al. (2011) Soluble CD163 made by monocyte/macrophages is a novel marker of HIV activity in early and chronic infection prior to and after anti-retroviral therapy. J. Infect. Dis. 204: 154-163

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