Overview

  • Product name
    Anti-IL-3RA / CD123 antibody [6H6]
    See all IL-3RA / CD123 primary antibodies
  • Description
    Mouse monoclonal [6H6] to IL-3RA / CD123
  • Host species
    Mouse
  • Tested applications
    Suitable for: ICC/IF, IP, Flow Cyt, WB, IHC-Frmore details
  • Species reactivity
    Reacts with: Human
  • Immunogen

    Tissue, cells or virus corresponding to IL-3RA/ CD123.

  • General notes

    Previously labelled as IL3RA. 

Properties

  • Form
    Liquid
  • Storage instructions
    Shipped at 4°C. Store at +4°C short term (1-2 weeks). Store at -20°C or -80°C. Avoid freeze / thaw cycle.
  • Storage buffer
    Preservative: 0.09% Sodium Azide
    Constituents: PBS, pH 7.2
  • Concentration information loading...
  • Purity
    Protein G purified
  • Clonality
    Monoclonal
  • Clone number
    6H6
  • Isotype
    IgG1
  • Light chain type
    kappa
  • Research areas

Applications

Our Abpromise guarantee covers the use of ab21562 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
ICC/IF Use at an assay dependent concentration.
IP Use at an assay dependent concentration.
Flow Cyt Use 1µg for 106 cells.

100 µl volume or 100 µl of whole blood.

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

WB Use at an assay dependent concentration. Detects a band of approximately 70 kDa.
IHC-Fr Use at an assay dependent concentration.

Target

  • Function
    This is a receptor for interleukin-3.
  • Sequence similarities
    Belongs to the type I cytokine receptor family. Type 5 subfamily.
  • Domain
    The WSXWS motif appears to be necessary for proper protein folding and thereby efficient intracellular transport and cell-surface receptor binding.
    The box 1 motif is required for JAK interaction and/or activation.
  • Cellular localization
    Membrane.
  • Information by UniProt
  • Database links
  • Alternative names
    • CD123 antibody
    • CD123 antigen antibody
    • hIL 3Ra antibody
    • hIL-3Ra antibody
    • hIL3Ra antibody
    • IL 3 receptor alpha SP2 isoform antibody
    • IL 3R alpha antibody
    • IL-3 receptor subunit alpha antibody
    • IL-3R subunit alpha antibody
    • IL-3R-alpha antibody
    • IL-3RA antibody
    • IL3 Receptor alpha antibody
    • IL3R antibody
    • Il3ra antibody
    • IL3RA_HUMAN antibody
    • IL3RAX antibody
    • IL3RAY antibody
    • IL3RX antibody
    • IL3RY antibody
    • Interleukin 3 receptor alpha chain antibody
    • Interleukin 3 receptor, alpha (low affinity) antibody
    • Interleukin 3 receptor, alpha antibody
    • Interleukin-3 Receptor alpha antibody
    • Interleukin-3 receptor subunit alpha antibody
    • Interleukin3 receptor antibody
    • Interleukin3 receptor, Y-chromosomal antibody
    see all

Images

  • Human peripheral blood lymphocytes stained with ab21562 (y-axis) and CD45RO PE/Cy5® (x-axis)

  • Immunofluorescence analysis of Human peripheral blood mononuclear cells, staining IL3RA / CD123 with ab21562.

    Cells were fixed with paraformaldehyde, permeabilized with 0.1% Triton X-100 in PBS and blocked with 1% BSA for 1 hour at 25&degC. Samples were incubated with primary antibody (1/100 in 1% BSA in PBS) for 24 hours at 4&degC. An AlexaFluor®488-conjugated goat anti-mouse polyclonal IgG (1/250) was used as the secondary antibody.

    See Abreview

  • Anti-IL-3RA / CD123 antibody [6H6] (ab21562) at 1/1000 dilution + Mouse spleen whole cell extract at 100 µg with Milk, 30 minutes at 25°C at 5 %

    Secondary
    Goat anti-mouse IgG (H+L), HRP conjugate at 1/4000 dilution

    Developed using the ECL technique.

    Exposure time: 15 minutes

References

This product has been referenced in:
  • Herling M  et al. TCL1 expression in plasmacytoid dendritic cells (DC2s) and the related CD4+ CD56+ blastic tumors of skin. Blood 101:5007-9 (2003). Read more (PubMed: 12576313) »
See 1 Publication for this product

Customer reviews and Q&As

1-10 of 10 Abreviews or Q&A

Abcam has not validated the combination of species/application used in this Abreview.
Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Human Tissue sections (normal spleen)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: EDTA pH7.8
Permeabilization
No
Specification
normal spleen
Blocking step
Casein as blocking agent for 5 minute(s) · Concentration: 2% · Temperature: RT°C
Fixative
Formaldehyde

Abcam user community

Verified customer

Submitted Sep 20 2016

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunocytochemistry/ Immunofluorescence
Sample
Human Cell (Human head & neck squamous cell carcinoma)
Permeabilization
Yes - Triton X-100 (0.1%) for 5 min
Specification
Human head & neck squamous cell carcinoma
Blocking step
BSA as blocking agent for 30 minute(s) · Concentration: 1% · Temperature: 22°C
Fixative
Paraformaldehyde

Abcam user community

Verified customer

Submitted Jan 30 2013

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunocytochemistry/ Immunofluorescence
Sample
Human Cell (Human peripheral blood mononuclear cells)
Permeabilization
Yes - 0.1% Triton X-100 in PBS
Specification
Human peripheral blood mononuclear cells
Blocking step
BSA as blocking agent for 30 minute(s) · Concentration: 1% · Temperature: 25°C
Fixative
Paraformaldehyde

Abcam user community

Verified customer

Submitted Jan 28 2013

Application
Western blot
Sample
Mouse Cell lysate - whole cell (Mouse spleen whole extract)
Gel Running Conditions
Reduced Denaturing (Tris-Glycine -10%)
Loading amount
100 µg
Specification
Mouse spleen whole extract
Blocking step
Milk as blocking agent for 30 minute(s) · Concentration: 5% · Temperature: 25°C

Abcam user community

Verified customer

Submitted Oct 30 2012

Application
Western blot
Sample
Mouse Cell lysate - whole cell (Mouse spleen whole extract)
Gel Running Conditions
Reduced Denaturing (10)
Loading amount
100 µg
Specification
Mouse spleen whole extract
Blocking step
Milk as blocking agent for 30 minute(s) · Concentration: 5% · Temperature: 25°C

Abcam user community

Verified customer

Submitted Oct 30 2012

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Flow Cytometry
Sample
Human Cell (PBMCs)
Permeabilization
No
Gating Strategy
FSC/SSC lymphocytes and then FSC-A/FSC-H singlets
Specification
PBMCs
Preparation
Cell harvesting/tissue preparation method: Ficoll
Sample buffer: PBS
Fixation
Paraformaldehyde

Abcam user community

Verified customer

Submitted Oct 01 2012

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunohistochemistry (Frozen sections)
Sample
Human Tissue sections (tonsil)
Permeabilization
No
Specification
tonsil
Blocking step
Serum as blocking agent for 30 minute(s) · Concentration: 6% · Temperature: 25°C
Fixative
Acetone

Abcam user community

Verified customer

Submitted Sep 28 2012

Answer

Thank you for your enquiry and your interest in our products.

Generally, each antibody and batch should be titrated to determine the optimal working dilutions/concentrations.

ab17051: this antibody has only been tested in ICC/IF and IHC/Fr but it is unsuitable for formalin-fixed paraffin-embedded sections. The unit size is 50 µg at 0.1mg/ml and it is suggested to apply for IHC-Fr at aconcentration of 4 µg/ml (please read application notes).

ab21562:this antibody has only been tested in IHC/Fr but it is unsuitable for formalin-fixed paraffin-embedded sections. The unit size is 100 µg at 0.5mg/ml but the working dilution needs to be optimized by the end user.

If you need any further assistance in the future, please do not hesitate to contact me.

Read More

Question
Answer

DISCOUNT CODE: *** Expiration date: January 30, 2012 Thanks for your call today and for your interest in ab21562. This code will give you 1 free primary antibody before the expiration date. To redeem this offer, please submit an Abreview for ab21562 in IHC-P and include this code in the “Additional Comments” section so we know the Abreview is for this promotion. For more information on how to submit an Abreview, please visit the site: www.abcam.com/Abreviews. The code will be active once the Abreview has been submitted and can be redeemed in one of the following ways: 1) Call to place your order and mention the code to our customer service department; 2) Include the code in your fax order; 3) Place your order on the web and enter the promotional code. Any feedback that you can provide will be greatly appreciated, whether positive or negative. If you have any further questions, please do not hesitate to contact us. We look forward to receiving your Abreview and wish you luck with your research. The terms and conditions applicable to this offer can be found here: www.abcam.com/collaborationdiscount.

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Answer

Thank you for your enquiry. I have added a paper to the antibody datasheet that uses the 6H6 antibody to stain tissue sections. This paper has used paraformaldehyde as a fixative. It does not look like they used antigen retrieval (although I was unable to get the detailed method they used for staining in reference #21). I would recommend that you take a look at the methods in reference #21 as well as this paper. A good positive tissue for staining would be tonsil. We typically recommend that the customer use a dose titration for staining ranging from 2.5 ug/ml to 20 ug/ml to establish optimal conditions. I hope this information is helpful to you. Please let me know if you need additional information.

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Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

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