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Alexa Fluor® 647 anti-human CD163 Antibody

Pricing & Availability
Clone
GHI/61 (See other available formats)
Regulatory Status
RUO
Workshop
VI M38
Other Names
GHI/61, M130, RM3/1, p155, Hemoglobin/Haptoglobin Complex Receptor, macrophage-associated antigen, ED2(rat)
Isotype
Mouse IgG1, κ
Ave. Rating
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Product Citations
publications
1_GHIslash61_AF647_CD163_Antibody_FC_052314.jpg
Human peripheral blood monocytes were stained with CD163 (clone GHI/63) Alexa Fluor® 647 (filled histogram) or mouse IgG1, κ Alexa Fluor® 647 isotype control (open histogram).
  • 1_GHIslash61_AF647_CD163_Antibody_FC_052314.jpg
    Human peripheral blood monocytes were stained with CD163 (clone GHI/63) Alexa Fluor® 647 (filled histogram) or mouse IgG1, κ Alexa Fluor® 647 isotype control (open histogram).
  • 2_11_Human_LN_CD163_CD4
    Confocal image of human lymph node sample acquired using the IBEX method of highly multiplexed antibody-based imaging: CD163 (red) in Cycle 3 and CD4 (blue) in Cycle 5. Tissues were prepared using ~1% (vol/vol) formaldehyde and a detergent. Following fixation, samples are immersed in 30% (wt/vol) sucrose for cryoprotection. Images are courtesy of Drs. Andrea J. Radtke and Ronald N. Germain of the Center for Advanced Tissue Imaging (CAT-I) in the National Institute of Allergy and Infectious Diseases (NIAID, NIH).
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333619 25 tests £81
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333620 100 tests £213
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Description

CD163 is a member of the group B scavenger receptor cysteine-rich superfamily, also known as GHI/61, M130, RM3/1, p155, hemoglobin-haptoglobin complex receptor, or macrophage-associated antigen. It is a 134 kD (non-reduced)/155 kD (reduced) glycoprotein primarily expressed on macrophages, Kupffer cells, monocytes, a subset of dendritic cells, and a subset of hematopoietic stem/progenitor cells. CD163 binds to haptoglobin-hemoglobin complex and TWEAK, and plays a role in clearing hemoglobin and regulating cytokine production by macrophages. Membrane CD163 can be cleaved by metalloproteinases (MMP), resulting in a soluble form. Elevated serum level of sCD163 has been implicated in many kinds of inflammatory diseases.

Product Details
Technical Data Sheet (pdf)

Product Details

Verified Reactivity
Human, Cynomolgus, Rhesus
Reported Reactivity
African Green
Antibody Type
Monoclonal
Host Species
Mouse
Formulation
Phosphate-buffered solution, pH 7.2, containing 0.09% sodium azide and BSA (origin USA)
Preparation
The antibody was purified by affinity chromatography and conjugated with Alexa Fluor® 647 under optimal conditions.
Concentration
Lot-specific (to obtain lot-specific concentration and expiration, please enter the lot number in our Certificate of Analysis online tool.)
Storage & Handling
The antibody solution should be stored undiluted between 2°C and 8°C, and protected from prolonged exposure to light. Do not freeze.
Application

FC - Quality tested
SB - Reported in the literature, not verified in house
Recommended Usage

Each lot of this antibody is quality control tested by immunofluorescent staining with flow cytometric analysis. For flow cytometric staining, the suggested use of this reagent is 5 µl per million cells in 100 µl staining volume or 5 µl per 100 µl of whole blood.

* Alexa Fluor® 647 has a maximum emission of 668 nm when it is excited at 633 nm / 635 nm.


Alexa Fluor® and Pacific Blue™ are trademarks of Life Technologies Corporation.

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Excitation Laser
Red Laser (633 nm)
Application Notes

Clone GHI/61 binds to domain 7 of CD163. Additional reported applications (for the relevant formats) include: immunocytochemical staining, immunoprecipitation, western blot1, and spatial biology (IBEX)6,7.

Additional Product Notes

Iterative Bleaching Extended multi-pleXity (IBEX) is a fluorescent imaging technique capable of highly-multiplexed spatial analysis. The method relies on cyclical bleaching of panels of fluorescent antibodies in order to image and analyze many markers over multiple cycles of staining, imaging, and, bleaching. It is a community-developed open-access method developed by the Center for Advanced Tissue Imaging (CAT-I) in the National Institute of Allergy and Infectious Diseases (NIAID, NIH).

Application References

(PubMed link indicates BioLegend citation)
  1. Pulford K, et al. 1992. Immunology 75:588. (ICC, IP, WB)
  2. Law SK, et al. 1993. Eur. J. Immunol. 23:2320.
  3. Madsen M, et al. 2004. J. Biol. Chem. 279:51561.
  4. Kim WK, et al. 2006. Am. J. Pathol. 168:822. (FC)
  5. Buttari B, et al. 2011. Atherosclerosis. 215:316. PubMed
  6. Radtke AJ, et al. 2020. Proc Natl Acad Sci USA. 117:33455-33465. (SB) PubMed
  7. Radtke AJ, et al. 2022. Nat Protoc. 17:378-401. (SB) PubMed
Product Citations
  1. Takiguchi H, et al. 2021. Scientific Reports. 11(1):8282. PubMed
  2. Smith MH, et al. 2023. Nat Immunol. . PubMed
  3. Kubala MH, et al. 2018. Cell Rep. 25:2177. PubMed
RRID
AB_2563475 (BioLegend Cat. No. 333619)
AB_2563475 (BioLegend Cat. No. 333620)

Antigen Details

Structure
134 kD (non-reduced)/155 kD (reduced) glycoprotein, scavenger receptor superfamily
Distribution

Monocytes, macrophages, Kuffer cells, subset of dendritic cells, subset of hematopoietic stem/progenitor cells

Function
Clearance of haptoglobin-hemoglobin complex, regulation of cytokine production by macrophages
Ligand/Receptor
Haptoglobin-hemoglobin complex, TWEAK
Cell Type
Dendritic cells, Hematopoietic stem and progenitors, Macrophages, Monocytes
Biology Area
Cell Biology, Immunology, Innate Immunity, Neuroscience, Neuroscience Cell Markers
Molecular Family
CD Molecules
Antigen References

1. Roth J, et al. 1994 Transolantation. 57:127
2. Van den Heuvel MM, et al.1999 J. Leukoc. Biol. 66:858
3. Sulahian TH, et al. 2000 Cytokines 12:1312
4. Fabriek BO, et al. 2007 J. Neuroimmunol. 187:179

Gene ID
9332 View all products for this Gene ID
UniProt
View information about CD163 on UniProt.org

Related FAQs

If an antibody clone has been previously successfully used in IBEX in one fluorescent format, will other antibody formats work as well?

It’s likely that other fluorophore conjugates to the same antibody clone will also be compatible with IBEX using the same sample fixation procedure. Ultimately a directly conjugated antibody’s utility in fluorescent imaging and IBEX may be specific to the sample and microscope being used in the experiment. Some antibody clone conjugates may perform better than others due to performance differences in non-specific binding, fluorophore brightness, and other biochemical properties unique to that conjugate.

Will antibodies my lab is already using for fluorescent or chromogenic IHC work in IBEX?

Fundamentally, IBEX as a technique that works much in the same way as single antibody panels or single marker IF/IHC. If you’re already successfully using an antibody clone on a sample of interest, it is likely that clone will have utility in IBEX. It is expected some optimization and testing of different antibody fluorophore conjugates will be required to find a suitable format; however, legacy microscopy techniques like chromogenic IHC on fixed or frozen tissue is an excellent place to start looking for useful antibodies.

Are other fluorophores compatible with IBEX?

Over 18 fluorescent formats have been screened for use in IBEX, however, it is likely that other fluorophores are able to be rapidly bleached in IBEX. If a fluorophore format is already suitable for your imaging platform it can be tested for compatibility in IBEX.

The same antibody works in one tissue type but not another. What is happening?

Differences in tissue properties may impact both the ability of an antibody to bind its target specifically and impact the ability of a specific fluorophore conjugate to overcome the background fluorescent signal in a given tissue. Secondary stains, as well as testing multiple fluorescent conjugates of the same clone, may help to troubleshoot challenging targets or tissues. Using a reference control tissue may also give confidence in the specificity of your staining.

How can I be sure the staining I’m seeing in my tissue is real?

In general, best practices for validating an antibody in traditional chromogenic or fluorescent IHC are applicable to IBEX. Please reference the Nature Methods review on antibody based multiplexed imaging for resources on validating antibodies for IBEX.

Go To Top Version: 4    Revision Date: 04/19/2022

For Research Use Only. Not for diagnostic or therapeutic use.

 

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This data display is provided for general comparisons between formats.
Your actual data may vary due to variations in samples, target cells, instruments and their settings, staining conditions, and other factors.
If you need assistance with selecting the best format contact our expert technical support team.

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