Product nameAnti-IRAK-1 antibody - BSA and Azide free
See all IRAK-1 primary antibodies
DescriptionRabbit polyclonal to IRAK-1 - BSA and Azide free
SpecificityNo cross reactivity to IRAK-2.
Tested applicationsSuitable for: WB, IP, IHC-P, Flow Cyt, ICC/IFmore details
Species reactivityReacts with: Mouse, Rat, Human, Rhesus monkey
Synthetic peptide corresponding to Human IRAK-1 aa 700-712 (C terminal).
(Peptide available as
- THP-1 or HeLa whole cell lysate.
Storage instructionsShipped at 4°C. Store at +4°C.
Storage bufferPreservative: 0.02% Sodium azide
Concentration information loading...
Purification notesAffinity chromatography purified via peptide column.
Immunizing Peptide (Blocking)
Our Abpromise guarantee covers the use of ab238 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|WB||1/1000. Predicted molecular weight: 80 kDa.Can be blocked with IRAK-1 peptide (ab6230). (THP-1 or HeLa whole cell) from non-activated cells.|
|IP||Use a concentration of 5 µg/ml.|
|IHC-P||Use a concentration of 1 µg/ml. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.|
|Flow Cyt||Use at an assay dependent concentration.
ab171870 - Rabbit polyclonal IgG, is suitable for use as an isotype control with this antibody.
|ICC/IF||Use a concentration of 10 µg/ml.|
FunctionBinds to the IL-1 type I receptor following IL-1 engagement, triggering intracellular signaling cascades leading to transcriptional up-regulation and mRNA stabilization. Isoform 1 binds rapidly but is then degraded allowing isoform 2 to mediate a slower, more sustained response to the cytokine. Isoform 2 is inactive suggesting that the kinase activity of this enzyme is not required for IL-1 signaling. Once phosphorylated, IRAK1 recruits the adapter protein PELI1.
Tissue specificityIsoform 1 and isoform 2 are ubiquitously expressed in all tissues examined, with isoform 1 being more strongly expressed than isoform 2.
Sequence similaritiesBelongs to the protein kinase superfamily. TKL Ser/Thr protein kinase family. Pelle subfamily.
Contains 1 protein kinase domain.
modificationsAutophosphorylated or is transphosphorylated by IRAK4 following recruitment to the IL-1RI. In the case of isoform 1, this is linked to ubiquitination and degradation.
Polyubiquitinated; after cell stimulation with IL-1-beta. Polyubiquitination occurs with polyubiquitin chains linked through 'Lys-63'.
- Information by UniProt
- AA48924 antibody
- Il1rak antibody
- Interleukin 1 receptor associated kinase 1 antibody
All lanes : Anti-IRAK-1 antibody - BSA and Azide free (ab238) at 1/2000 dilution
Lane 1 : THP-1 (THP)whole cell lysate
Lane 2 : HeLa (HL) whole cell lysate
Predicted band size: 80 kDa
Observed band size: 80 kDa
ab238 at 10µg/ml staining Hela cells by ICC/IF
IHC image of ab238 staining in human breast carcinoma formalin fixed paraffin embedded tissue section, performed on a Leica BondTM system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab238, 1µg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.
For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.
ab238 staining IRAK-1 in Human platelet cells by Flow cytometry.
Cells were fixed in paraformaldehyde and permeabilized using 0.1% Triton-X-100 in 2% BSA for 15 minutes. Primary antibody used at a 1/200 dilution and incubated for 16 hours at 4°C. The secondary antibody used was an Alexa Fluor®488 conjugated chicken anti-rabbit IgG (H+L) at a 1/500 dilution.
P : Permeabilized US : Unstained (Red Peak) IGG RB : IgG Rabbit (Blue Peak) IRAK Ab (Green Peak)
ab238 staining IRAK-1 in murine RAW 264.7 cells by Immunocytochemistry/ Immunofluorescence.Cells were fixed in paraformaldehyde, permeabilized using 0.1% Triton-X100 in 2% BSA for 15 minutes, blocked with 2% BSA for 1 hour at 22°C and then incubated with ab238 at a 1/150 dilution for 16 hours at 4°C. The secondary used was an Alexa-Fluor 488 conjugated chicken anti-rabbit IgG (H+L) used at a 1/1000 dilution.
IRAK was immunoprecipitated using 0.5mg Hela whole cell extract, 5µg of Rabbit polyclonal to IRAK-1 and 50µl of protein G magnetic beads (+). No antibody was added to the control (-).
The antibody was incubated under agitation with Protein G beads for 10min, Hela whole cell extract lysate diluted in RIPA buffer was added to each sample and incubated for a further 10min under agitation.
Proteins were eluted by addition of 40µl SDS loading buffer and incubated for 10min at 70oC; 10µl of each sample was separated on a SDS PAGE gel, transferred to a nitrocellulose membrane, blocked with 5% BSA and probed with ab238.
Secondary: Mouse monoclonal [SB62a] Secondary Antibody to Rabbit IgG light chain (HRP) (ab99697).
Band: 80kDa: IRAK-1
Immunofluorescence of IRAK-1 in HeLa cells using ab238 at 20 ug/ml.
This product has been referenced in:
- Chen Y et al. MicroRNA-146a-5p attenuates irradiation-induced and LPS-induced hepatic stellate cell activation and hepatocyte apoptosis through inhibition of TLR4 pathway. Cell Death Dis 9:22 (2018). WB ; Mouse . Read more (PubMed: 29348414) »
- Wang Z et al. Chronic constriction injury-induced microRNA-146a-5p alleviates neuropathic pain through suppression of IRAK1/TRAF6 signaling pathway. J Neuroinflammation 15:179 (2018). WB, IHC (PFA fixed) ; Rat . Read more (PubMed: 29885668) »