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PE anti-human CD123 Antibody

Pricing & Availability
Clone
6H6 (See other available formats)
Regulatory Status
RUO
Other Names
IL-3Rα, IL-3 Receptor alpha
Isotype
Mouse IgG1, κ
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Product Citations
publications
6H6_PE_CD123_Antibody_1_091924
Human peripheral blood lymphocytes stained with 6H6 PE and CD45RO PE/Cyanine5
  • 6H6_PE_CD123_Antibody_1_091924
    Human peripheral blood lymphocytes stained with 6H6 PE and CD45RO PE/Cyanine5
  • 6H6_PE_CD123_Antibody_2_101024
    Multiplexed IHC staining of PE anti-CD123 (clone 6H6) on formalin-fixed paraffin-embedded human tonsil tissue, validated for use on the Cellscape™. The tissue was iteratively stained with PE anti-CD123 (clone 6H6, green) and Alexa Fluor® 488 anti-CD3 (red) for one hour at room temperature. Nuclei were counterstained with Hoechst 33342. Images were captured with a 20X objective. Scale bar: 50 µm
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306005 25 tests $82
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306006 100 tests $175
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Description

CD123 is the 70 kD transmembrane α chain of the IL-3 receptor. Alone, CD123 binds IL-3 with low affinity; when CD123 associates with CD131 (common β chain), it binds IL-3 with high affinity. CD123 does not transduce intracellular signals upon binding IL-3 and requires the β chain for this function. CD123 is expressed by myeloid precursors, macrophages, dendritic cells, mast cells, basophils, megakaryocytes, and some B cells.

Product Details
Technical data sheet

Product Details

Verified Reactivity
Human
Reported Reactivity
Rhesus
Antibody Type
Monoclonal
Host Species
Mouse
Immunogen
Human IL-3Rα transfected COS cells.
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 PE 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 - Community Verified
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.

Excitation Laser
Blue Laser (488 nm)
Green Laser (532 nm)/Yellow-Green Laser (561 nm)
Application Notes

Clone 6H6 does not inhibit IL-3 binding to low- or high-affinity IL-3Rs. Additional reported applications (for the relevant formats) include: Western blotting1, immunoprecipitation1, and immunohistochemical staining of acetone-fixed frozen sections2 and also paraformaldehyde fixed paraffin embedded tissue7.

Additional Product Notes

For the use of this antibody in spatial biology (SB), we have partnered with Bruker Spatial Biology Biosciences for demonstration of this antibody on their next-generation ChipCytometry instrument called the CellScape™. The CellScape platform is an end-to-end solution for highly multiplexed spatial omics. Combining an advanced, purpose-built imaging system with easy-to-use fluidics for walk-away automation, the CellScape system will accelerate your exploration into the rapidly evolving field of spatial biology. More information on the the Bruker Spatial Biology CellScape and a complete list of our antibodies that have been demonstrated on the instrument can be found here.

Application References

(PubMed link indicates BioLegend citation)
  1. Sun Q, et al. 1996. Blood 87:83. (IP, WB)
  2. Herling M, et al. 2003. Blood 101:5007. (IHC)
  3. Charles N, et al. 2010. Nat. Med. 16:701. (FC) PubMed
  4. Martin-Gayo E, et al. 2010. Blood 115:5366. PubMed
  5. Chen SC, et al. 2010. Arch Dermatol Res. 302:113. PubMed
  6. Liu Y, et al. 2012. Food Chem Toxicol. 50:1920. PubMed
  7. Peduzzi E, et al. 2007. J. Invest. Dermatol. 127:638. (IHC)
Product Citations
  1. Zaro BW, et al. 2019. J Immunol. 202:2737. PubMed
  2. Luchsinger LL, et al. 2020. Cell Stem Cell. 25(2):225-240. PubMed
  3. Randrianarison-Huetz V, et al. 2010. Blood. 115:2784. PubMed
  4. Li M, et al. 2023. Front Immunol. 14:1087923. PubMed
  5. Woo MS, et al. 2021. iScience. 24(7):102752. PubMed
  6. Sefik E, et al. 2022. Nature. 606:585. PubMed
  7. Shahid AM, et al. 2022. PLoS One. 17:e0278209. PubMed
  8. Baudet A, et al. 2022. Bio Protoc. 12:e4353. PubMed
  9. Han L, et al. 2012. PLoS One. 4:e7989. PubMed
  10. Jarosch S, et al. 2022. STAR Protoc. 3:101374. PubMed
  11. Kim ST, et al. 2022. Nat Commun. 13:1970. PubMed
  12. Jarosch S, et al. 2021. Cell Rep Methods. 1:100104. PubMed
  13. Warmuth S, et al. 2022. Oncoimmunology. 10:2004661. PubMed
  14. Sefik E, et al. 2021. Nat Biotechnol. . PubMed
  15. Houtsma R, et al. 2021. STAR Protoc. 2:100864. PubMed
  16. Le J, et al. 2020. Immunity. 52(6):1105-1118.e9. PubMed
  17. Felce JH, et al. 2018. Sci Signal. 11:eaat0756. PubMed
  18. Li M, et al. 2020. Nat Commun. 4051:11. PubMed
  19. Kerdidani D, et al. 2022. J Exp Med. 219:. PubMed
  20. Leylek R, et al. 2020. Cell Rep. 32:108180. PubMed
RRID
AB_314580 (BioLegend Cat. No. 306005)
AB_314580 (BioLegend Cat. No. 306006)

Antigen Details

Structure
Ig superfamily, type I transmembrane glycoprotein, associates with CDw131, 70 kD
Distribution

Myeloid precursors, basophils, mast cells, macrophages, dendritic cells, megakaryocytes, subset of lymphocytes

Function
Hematopoietic cell proliferation, differentiation
Ligand/Receptor
IL-3
Cell Type
Basophils, Dendritic cells, Hematopoietic stem and progenitors, Lymphocytes, Macrophages, Mast cells, Megakaryocytes
Biology Area
Immunology
Molecular Family
CD Molecules, Cytokine/Chemokine Receptors
Antigen References

1. Miyajima A, et al. 1993. Blood 82:1960.

Gene ID
3563 View all products for this Gene ID
UniProt
View information about CD123 on UniProt.org

Related FAQs

What type of PE do you use in your conjugates?
We use R-PE in our conjugates.
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: 3    Revision Date: 10/10/2024

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

 

This product is supplied subject to the terms and conditions, including the limited license, located at www.biolegend.com/terms) ("Terms") and may be used only as provided in the Terms. Without limiting the foregoing, BioLegend products may not be used for any Commercial Purpose as defined in the Terms, resold in any form, used in manufacturing, or reverse engineered, sequenced, or otherwise studied or used to learn its design or composition without express written approval of BioLegend. Regardless of the information given in this document, user is solely responsible for determining any license requirements necessary for user’s intended use and assumes all risk and liability arising from use of the product. BioLegend is not responsible for patent infringement or any other risks or liabilities whatsoever resulting from the use of its products.

 

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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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