Recombinant Anti-IL3RA/CD123 antibody [EPR24481-20] (ab280355)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR24481-20] to IL3RA/CD123
- Suitable for: IP, Flow Cyt, WB, ICC/IF, IHC-P
- Reacts with: Human
Related conjugates and formulations
Overview
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Product name
Anti-IL3RA/CD123 antibody [EPR24481-20]
See all IL3RA/CD123 primary antibodies -
Description
Rabbit monoclonal [EPR24481-20] to IL3RA/CD123 -
Host species
Rabbit -
Tested applications
Suitable for: IP, Flow Cyt, WB, ICC/IF, IHC-Pmore details -
Species reactivity
Reacts with: Human -
Immunogen
Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.
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Positive control
- WB: Human tonsil, HDLM-2, KG-1a lysates. IHC-P: Human tonsil and Hodgkin's lymphoma tissues. ICC/IF: HDLM-2 cells. Flow Cyt: HDLM-2, Human peripheral blood mononuclear cells. IP: HDLM-2 cells
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General notes
This product is a recombinant monoclonal antibody, which offers several advantages including:
- - High batch-to-batch consistency and reproducibility
- - Improved sensitivity and specificity
- - Long-term security of supply
- - Animal-free production
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle. -
Storage buffer
Preservative: 0.01% Sodium azide
Constituents: 59.94% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA -
Concentration information loading...
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Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR24481-20 -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
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Compatible Secondaries
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Dyes/Markers
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Isotype control
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Related Products
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab280355 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 |
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IP |
1/30.
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Flow Cyt |
1/500.
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WB |
1/1000. Predicted molecular weight: 43 kDa.
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ICC/IF |
1/1000.
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IHC-P |
1/500.
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Notes |
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IP
1/30. |
Flow Cyt
1/500. |
WB
1/1000. Predicted molecular weight: 43 kDa. |
ICC/IF
1/1000. |
IHC-P
1/500. |
Target
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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
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Database links
- Entrez Gene: 3563 Human
- Omim: 308385 Human
- Omim: 430000 Human
- SwissProt: P26951 Human
- Unigene: 632790 Human
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Alternative names
- CD123 antibody
- CD123 antigen antibody
- hIL 3Ra antibody
see all
Images
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All lanes : Anti-IL3RA/CD123 antibody [EPR24481-20] (ab280355) at 1/1000 dilution
Lane 1 : Human tonsil tissue lysate at 40 µg
Lane 2 : HDLM-2 (human hodgkin lymphoma ) whole cell lysate at 20 µg
Lane 3 : K-562 (human chronic myelogenous leukemia lymphoblast) whole cell lysate at 20 µg
Secondary
Lane 1 : Goat Anti-Rabbit IgG (HRP) with minimal cross-reactivity with human IgG at 1/2000 dilution
Lanes 2-3 : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/50000 dilution (Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated)
Predicted band size: 43 kDa
Observed band size: 60 kDa why is the actual band size different from the predicted?Blocking and diluting buffer and concentration: 5% NFDM/TBST
The expression profile/molecular weight observed is consistent with what has been described in the literature (PMID:30647406).
Negative control: K-562 cell (PMID: 30647406)
Exposure time: 37 seconds
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-IL3RA/CD123 antibody [EPR24481-20] (ab280355)
Immunohistochemical analysis of paraffin-embedded human tonsil tissue labeling IL3RA/CD123 with ab280355 at 1/500 dilution followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Positive staining on human tonsil (PMID: 27340692, PMID:15728234, PMID:28205524). The section was incubated with ab280355 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin.
Secondary antibody only control: Secondary antibody is a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).
Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.
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Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HDLM-2 cells labelling IL3RA/CD123 with ab280355 at 1/1000 dilution, followed by ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) antibody at 1/1000 dilution (Green). Confocal image showing membranous staining in HDLM-2 cells.
Negative control: K-562 (PMID: 26531164).
ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) at a 1/200 dilution (Red). The nuclear counterstain was DAPI (Blue).
Secondary antibody only control: Secondary antibody is ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) at 1/1000 dilution.
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Flow cytometric analysis of K-562(Human chronic myelogenous leukemia lymphoblast, Left)/ HDLM-2(Human Hodgkin lymphoma, Right) cells labelling IL3RA/CD123 with ab280355 at 1/500 dilution (0.1ug)/ red compared with a Rabbit monoclonal IgG (ab172730) (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) at 1/2000 dilution was used as the secondary antibody.
Negative control: K-562 cell (PMID: 26531164).
Gated on viable cells.
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IL3RA/CD123 was immunoprecipitated from 0.35 mg HDLM-2 (human hodgkin lymphoma ) whole cell lysate 10 µg with ab280355 at 1/30 dilution (2µg in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab280355 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP)(ab131366) was used at 1/5000 dilution.
Lane 1: HDLM-2 whole cell lysate 10 µg
Lane 2: ab280355 IP in HDLM-2 whole cell lysate
Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab280355 in HDLM-2 whole cell lysate
Blocking and dilution buffer and concentration: 5% NFDM/TBST.
Exposure time: 32 seconds
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Anti-IL3RA/CD123 antibody [EPR24481-20] (ab280355) at 1/1000 dilution + KG-1a (human bone marrow myeloblast) whole cell lysate at 20 µg
Secondary
Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution
Predicted band size: 43 kDa
Observed band size: 60 kDa why is the actual band size different from the predicted?Blocking and diluting buffer and concentration: 5% NFDM/TBST
The expression profile/molecular weight observed is consistent with what has been described in the literature (PMID:30647406).
Exposure time: 103 seconds
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-IL3RA/CD123 antibody [EPR24481-20] (ab280355)
Immunohistochemical analysis of paraffin-embedded human Hodgkin's lymphoma tissue labeling IL3RA/CD123 with ab280355 at 1/500 dilution followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Positive staining on human Hodgkin's lymphoma. The section was incubated with ab280355 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin.
Secondary antibody only control: Secondary antibody is a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).
Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.
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Flow cytometric analysis of human peripheral blood mononuclear cell (PBMC) cells labelling IL3RA/CD123 with ab280355 at 1/500 dilution (0.1ug)/ Right compared with a Rabbit monoclonal IgG (ab172730) / Left isotype control. Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) at 1/2000 dilution was used as the secondary antibody.
Cells were stained with rabbit IgG or ab280355. Then stained with anti-HLA-DR conjugated to PE.
Gated on viable cells.
Protocols
To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.
Datasheets and documents
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SDS download
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Datasheet download
Certificate of Compliance
References (0)
ab280355 has not yet been referenced specifically in any publications.