Recombinant Anti-IL-1RAcP antibody [EPR22678-67] (ab256461)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR22678-67] to IL-1RAcP
- Suitable for: WB, Flow Cyt, IP, Indirect ELISA
- Knockout validated
- Reacts with: Human
Related conjugates and formulations
Overview
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Product name
Anti-IL-1RAcP antibody [EPR22678-67]
See all IL-1RAcP primary antibodies -
Description
Rabbit monoclonal [EPR22678-67] to IL-1RAcP -
Host species
Rabbit -
Tested applications
Suitable for: WB, Flow Cyt, IP, Indirect ELISAmore details
Unsuitable for: ICC/IF or IHC-P -
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: KARPAS-299 and HepG2 whole cell lysate. Flow Cyt: HeLa and HepG2 cells. IP: KARPAS-299 whole cell lysate.
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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
pH: 7.2
Preservative: 0.01% Sodium azide
Constituents: PBS, 40% Glycerol, 0.05% BSA -
Concentration information loading...
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Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR22678-67 -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
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Compatible Secondaries
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Isotype control
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KO cell lines
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Recombinant Protein
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Related Products
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab256461 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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WB |
1/1000. Detects a band of approximately 80 kDa (predicted molecular weight: 65 kDa).
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Flow Cyt |
1/60.
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IP |
1/30.
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Indirect ELISA |
Use at an assay dependent concentration.
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Notes |
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WB
1/1000. Detects a band of approximately 80 kDa (predicted molecular weight: 65 kDa). |
Flow Cyt
1/60. |
IP
1/30. |
Indirect ELISA
Use at an assay dependent concentration. |
Target
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Function
Coreceptor with IL1R1. Associates with IL1R1 bound to IL1B to form the high affinity interleukin-1 receptor complex which mediates interleukin-1-dependent activation of NF-kappa-B and other pathways. Signaling involves the recruitment of adapter molecules such as TOLLIP, MYD88, and IRAK1 or IRAK2 via the respective TIR domains of the receptor/coreceptor subunits. Recruits TOLLIP to the signaling complex. Does not bind to interleukin-1 alone; binding of IL1RN to IL1R1, prevents its association with IL1R1 to form a signaling complex. The cellular response is modulated through a non-signaling association with the membrane IL1R2 decoy receptor. Secreted forms (isoforms 2 and 3) associate with secreted ligand-bound IL1R2 and increase the affinity of secreted IL1R2 for IL1B; this complex formation may be the dominant mechanism for neutralization of IL1B by secreted/soluble receptors. -
Tissue specificity
Detected in liver, skin, placenta, thymus and lung. -
Sequence similarities
Belongs to the interleukin-1 receptor family.
Contains 3 Ig-like C2-type (immunoglobulin-like) domains.
Contains 1 TIR domain. -
Cellular localization
Secreted and Cell membrane. - Information by UniProt
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Database links
- Entrez Gene: 3556 Human
- Omim: 602626 Human
- SwissProt: Q9NPH3 Human
- Unigene: 478673 Human
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Alternative names
- C3orf13 antibody
- FLJ37788 antibody
- IL 1 receptor accessory protein antibody
see all
Images
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Flow cytometry overlay histogram showing wild-type HeLa (green line) and IL1RAP knockout HeLa cells (ab273375) stained with ab256461 (red line). The cells were incubated in 1x PBS containing 10% normal goat serum to block non-specific protein-protein interaction followed by the antibody (ab256461) (1x106 in 100μl at 10 μ/ml) for 30 min at 4°C.
The secondary antibody Goat anti-rabbit IgG H&L (Alexa Fluor® 488, pre-adsorbed) (ab150081) was used at 1/2000 for 30 min at 4°C.
Isotype control antibody was Rabbit IgG (monoclonal) (ab172730) used at the same concentration and conditions as the primary antibody (wild-type HeLa - black line; IL1RAP knockout HeLa - grey line). Unlabelled sample was also used as a control (this line is not shown for the purpose of simplicity).
Acquisition of >5000 events were collected using a 50 mW Blue laser (488nm) and 525/40 bandpass filter.
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All lanes : Anti-IL-1RAcP antibody [EPR22678-67] (ab256461) at 1/1000 dilution
Lane 1 : KARPAS-299 (human anaplastic large cell lymphoma) whole cell lysate
Lane 2 : Raji (human burkitt's lymphoma b lymphocyte) whole cell lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution
Predicted band size: 65 kDa
Exposure time: 3 minutesThe molecular weight observed is consistent with what has been described in the literature (PMID: 30514753).
Negative control: Raji (PMID: 30514753).
Blocking/Dilution buffer: 5% NFDM/TBST.
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Anti-IL-1RAcP antibody [EPR22678-67] (ab256461) at 1/1000 dilution + HepG2 (human hepatocellular carcinoma epithelial cell) whole cell lysate at 20 µg
Secondary
Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution
Predicted band size: 65 kDa
Exposure time: 3 minutesThe molecular weight observed is consistent with what has been described in the literature (PMID: 30514753).
This blot was developed using a higher sensitivity ECL substrate.
Blocking/Dilution buffer: 5% NFDM/TBT.
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Flow cytometric analysis of Raji (Human burkitt's lymphoma b lymphocyte, Left panel) and HepG2 (Human hepatocellular carcinoma epithelial cell, Right panel) cells labeling IL-1RAcP with ab256461 at a 1/60 dilution (1μg, red), compared with a Rabbit monoclonal IgG (ab172730) (Black) isotype control and an unlabeled control (cells without incubation with primary antibody and secondary antibody) (Blue). Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) at a 1/2000 dilution was used as the secondary antibody. Negative control: Raji (PMID: 30514753). Gated on viable cells.
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IL-1RAcP was immunoprecipitated from 0.35 mg KARPAS-299 (human anaplastic large cell lymphoma) whole cell lysate 10μg with ab256461 at 1/30 dilution (2μg in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab256461 at a 1/1000 dilution (0.59 μg/ml). VeriBlot for IP Detection Reagent (HRP) (ab131366) was used at 1/5000 dilution.
Lane 1: KARPAS-299 (human anaplastic large cell lymphoma) whole cell lysate 10μg.
Lane 2: ab256461 IP in KARPAS-299 whole cell lysate.
Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab256461 in KARPAS-299 whole cell lysate.
Blocking/Dilution buffer: 5% NFDM/TBST.
Exposure time: 75 secs.
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ELISA analysis of Hu IL1RAP recombinant protein at 1000 ng/mL with ab256461. An Alkaline Phosphatase-conjugated AffiniPure Goat Anti-Rabbit IgG (H+L) at 1/2500 dilution was used as the secondary antibody.
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 (2)
ab256461 has been referenced in 2 publications.
- Zhou J et al. Structural basis of the IL-1 receptor TIR domain-mediated IL-1 signaling. iScience 25:104508 (2022). PubMed: 35754719
- Wei W et al. IL-1 signaling pathway molecules as key markers in childhood asthma. Pediatr Allergy Immunol 32:305-313 (2021). PubMed: 33025692