Overview

  • Product name
    Anti-CD163 antibody [GHI/61]
    See all CD163 primary antibodies
  • Description
    Mouse monoclonal [GHI/61] to CD163
  • Host species
    Mouse
  • Tested applications
    Suitable for: WB, IHC-P, Flow Cytmore details
  • Species reactivity
    Reacts with: Rabbit, Human
  • Immunogen

    CD163 antibody was raised against spleen cells from a case of hairy cell leukemia.

  • Positive control
    • Human peripheral blood cells, Human Placenta, Human Spleen, and Human Liver tissues

Properties

Applications

Our Abpromise guarantee covers the use of ab111250 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes
WB Use at an assay dependent concentration. Predicted molecular weight: 125 kDa.
IHC-P Use a concentration of 20 µg/ml. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
Flow Cyt Use 0.5µg for 105-8 cells.

ab170190 - Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody.

 

Target

  • Function
    Acute phase-regulated receptor involved in clearance and endocytosis of hemoglobin/haptoglobin complexes by macrophages and may thereby protect tissues from free hemoglobin-mediated oxidative damage. May play a role in the uptake and recycling of iron, via endocytosis of hemoglobin/haptoglobin and subsequent breakdown of heme. Binds hemoglobin/haptoglobin complexes in a calcium-dependent and pH-dependent manner. Exhibits a higher affinity for complexes of hemoglobin and multimeric haptoglobin of HP*1F phenotype than for complexes of hemoglobin and dimeric haptoglobin of HP*1S phenotype. Induces a cascade of intracellular signals that involves tyrosine kinase-dependent calcium mobilization, inositol triphosphate production and secretion of IL6 and CSF1. Isoform 3 exhibits the higher capacity for ligand endocytosis and the more pronounced surface expression when expressed in cells.
    After shedding, the soluble form (sCD163) may play an anti-inflammatory role, and may be a valuable diagnostic parameter for monitoring macrophage activation in inflammatory conditions.
  • Tissue specificity
    Expressed in monocytes and mature macrophages such as Kupffer cells in the liver, red pulp macrophages in the spleen, cortical macrophages in the thymus, resident bone marrow macrophages and meningeal macrophages of the central nervous system. Expressed also in blood. Isoform 1 is the lowest abundant in the blood. Isoform 2 is the lowest abundant in the liver and the spleen. Isoform 3 is the predominant isoform detected in the blood.
  • Sequence similarities
    Contains 9 SRCR domains.
  • Domain
    The SRCR domain 3 mediates calcium-sensitive interaction with hemoglobin/haptoglobin complexes.
  • Post-translational
    modifications
    A soluble form (sCD163) is produced by proteolytic shedding which can be induced by lipopolysaccharide, phorbol ester and Fc region of immunoglobulin gamma. This cleavage is dependent on protein kinase C and tyrosine kinases and can be blocked by protease inhibitors. The shedding is inhibited by the tissue inhibitor of metalloproteinase TIMP3, and thus probably induced by membrane-bound metalloproteinases ADAMs.
    Phosphorylated.
  • Cellular localization
    Secreted and Cell membrane. Isoform 1 and isoform 2 show a lower surface expression when expressed in cells.
  • Information by UniProt
  • Database links
  • Alternative names
    • C163A_HUMAN antibody
    • CD 163 antibody
    • CD163 antibody
    • CD163 antigen antibody
    • CD163 molecule antibody
    • Hemoglobin scavenger receptor antibody
    • M130 antibody
    • M130 antigen precursor antibody
    • Macrophage associated antigen antibody
    • MM130 antibody
    • OTTHUMP00000238617 antibody
    • OTTHUMP00000238618 antibody
    • OTTHUMP00000238619 antibody
    • OTTHUMP00000238620 antibody
    • SCARI1 antibody
    • Scavenger receptor cysteine rich type 1 protein M130 antibody
    • sCD163 antibody
    • Soluble CD163 antibody
    see all

Images

  • Staining of normal Human peripheral blood cells with 0.25 ug of Purified Mouse IgG1, K isotype control (open histogram) or 0.25 ug of ab111250 (colored histogram) followed by FITC Anti-Mouse IgG. Cells in the monocyte gate were used for analysis.
  • Negative control - immunohistochemical staining of paraffin-embedded formalin fixed human cerebral cortex using ab111250 at 20 µg/mL.

  • Immunohistochemical staining of paraffin-embedded formalin fixed human liver using ab111250 at 20 µg/mL.

  • Formalin-Fixed, Paraffin-Embedded Human Placenta tissue stained using ab111250 at a concentration of 20 µg/ml.
  • Formalin-Fixed, Paraffin-Embedded Human Spleen tissue stained using ab111250 at a concentration of 20 µg/ml.
  • Formalin-Fixed, Paraffin-Embedded Human Liver tissue stained using ab111250 at a concentration of 20 µg/ml.

References

This product has been referenced in:
  • Han L  et al. Effects of silibinin-loaded thermosensitive liposome-microbubble complex on inhibiting rabbit liver VX2 tumors in sub-hyperthermia fields. Exp Ther Med 15:1233-1240 (2018). Read more (PubMed: 29434709) »
  • Gregory KJ  et al. Regulation of early growth response 2 expression by secreted frizzled related protein 1. BMC Cancer 17:473 (2017). Read more (PubMed: 28687085) »
See all 5 Publications for this product

Customer reviews and Q&As

1-4 of 4 Abreviews or Q&A

Application
Immunohistochemistry (Frozen sections)
Sample
Rat Tissue sections (Brain)
Permeabilization
Yes - 0.1 M PBS with 3% Triton X
Specification
Brain
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 24°C
Fixative
Paraformaldehyde

Dr. Ruma Raha-Chowdhury

Verified customer

Submitted Aug 20 2015

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Blocking step
Protein Block serum free from Dako as blocking agent for 30 minute(s) · Concentration: 100% · Temperature: 37°C
Antigen retrieval step
Enzymatic
Sample
Rabbit Tissue sections (Ear wound exudates)
Specification
Ear wound exudates
Permeabilization
No
Fixative
Formaldehyde

Abcam user community

Verified customer

Submitted May 29 2013

Answer


IF by definition refers to any antibody application that utilizes fluorescence as the detection method. Thus, technically ICC and IF are one in the same, and you could argue that IHC is also a form of IF.

For the purposes of searching for products on our website, we distinguish those antibodies that have been tested in IHC with IHC followed by a letter designation (P-paraffin embedded sections, Fr-frozen sections, FoFr-formaldehyde fixed frozen sections). If an antibody has ICC or ICC/IF listed under tested applications they are the same thing and guaranteed to work in immunocytochemistry.

It sounds like you are performing IHC-P, immunohistochemistry on paraffin embedded sections; thus, all three antibodies that you identified, ab53003, ab8216, and ab111250 are tested and guaranteed to detect their respective targets via IHC-P in human samples. As stated in our AbPromise guarantee, we are happy to provide scientific support, replacement or refund should these products if they do not perform as indicated on the datasheet. More information on our Abpromise may be found at the following link:

https://www.abcam.com/index.html?pageconfig=abpromise

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Answer

Muchas gracias por tu respuesta.

Es importante que todos los anticuerpos que escojas para llevar a cabo IHC-P en tejido humano estén validados (y por tanto garantizados por Abcam) para funcionar en esta especie y aplicación. Esta información aparece en la hoja técnica de los productos.

También es importante echar un vistazo a las hojas de datos de cada uno de los productos para tener en cuenta recomendaciones especificas, tales como diluciones, métodos de recuperación antigénica, y demás que pueden aparecer en ellas. A las fichas técnicas se puede acceder desde la página web, escribiendo el código del anticuerpo (ab955, por ejemplo) en la casilla azul en la parte de arriba de nuestra página web (https://www.abcam.com/).

Desconozco las combinaciones que queréis llevar a cabo mediante inmunofluorescencia doble con los anticuerpos mencionados. En general, a la hora de hacer inmunoensayos múltiples hay que tener mucha precaución para evitar reactividad cruzada. Es importante usar anticuerpos procedentes de distintas especies si se puede, o al menos con diferentes isotipos. Una buena forma de evitar reactividad cruzada es usando secundarios de la misma especie. Además esto permite usar suero de dicha especie para bloquear.

Te copio los links a los protocolos que tenemos en nuestra web para llevar a cabo doble inmunofluorescencia por s pudieran resultar de ayuda:

https://www.abcam.com/index.html?pageconfig=resource&rid=11459

https://www.abcam.com/index.html?pageconfig=resource&rid=11458

De todas maneras si quieres que comentemos distintas posibilidades y escenarios posibles, no dudes en contactarme otra vez.

Respecto a los secundarios, tenemos muchos anticuerpos que podrían serviros. Te aconsejo que en función de los primarios y combinaciones que vayáis a llevar a cabo, optéis por uno u otro. Para llevar a cabo la búsqueda de los secundarios desde la pagina web (https://www.abcam.com/) pincha en la pestaña “Secondary Antibodies”. Se abre una nueva ventana de “Advance Search”, en la cual se pueden elegir las categorías requeridas. La única categoría obligatoria es el isotipo del anticuerpo, pues tiene que coincidir con el del primario. El resto de categorías son opcionales, pero son muy útiles para afinar la búsqueda y elegir el anticuerpo más apropiado según la especie del primario, la conjugación deseada, la clonalidad, la aplicación en la que se va a testar, etc…

Los precios y la disponibilidad pueden consultarse igualmente en la parte superior de la datasheet de los productos. Si necesitas un presupuesto no dudes en hacérmelo saber para que te lo envíe. Además tenemos algún descuento para pedidos superiores a 5 productos, por lo que si vais a realizar un pedido de estas características te animaría a que nos contactaras primero.

Espero que esta información sea de ayuda. Como te digo, para cualquier aclaración o más sugerencias, estaré encantada de echar una mano.

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