Overview

  • Product name
    Anti-IRAK-1 antibody - BSA and Azide free
    See all IRAK-1 primary antibodies
  • Description
    Rabbit polyclonal to IRAK-1 - BSA and Azide free
  • Host species
    Rabbit
  • Specificity

    No cross reactivity to IRAK-1.

  • Tested applications
    Suitable for: WB, IP, IHC-P, Flow Cyt, ICC/IFmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Human, Rhesus monkey
  • Immunogen

    Synthetic peptide corresponding to Human IRAK-1 aa 700-712 (C terminal).
    (Peptide available as ab6230)

  • Positive control
    • THP-1 or HeLa whole cell lysate.

Properties

Applications

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.

Application Abreviews Notes
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.

Target

  • Function
    Binds 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 specificity
    Isoform 1 and isoform 2 are ubiquitously expressed in all tissues examined, with isoform 1 being more strongly expressed than isoform 2.
  • Sequence similarities
    Belongs to the protein kinase superfamily. TKL Ser/Thr protein kinase family. Pelle subfamily.
    Contains 1 protein kinase domain.
  • Post-translational
    modifications
    Autophosphorylated 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
  • Database links
  • Alternative names
    • AA48924 antibody
    • Il1rak antibody
    • Interleukin 1 receptor associated kinase 1 antibody
    • Interleukin-1 receptor-associated kinase 1 antibody
    • IRAK antibody
    • IRAK-1 antibody
    • Irak1 antibody
    • IRAK1-S antibody
    • IRAK1_HUMAN antibody
    • mPLK antibody
    • OTTHUMP00000026014 antibody
    • OTTHUMP00000026015 antibody
    • OTTHUMP00000026020 antibody
    • OTTHUMP00000180621 antibody
    • Pelle antibody
    • Pelle homolog antibody
    • Pelle-like protein kinase antibody
    • Plpk antibody
    see all

Images

  • 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-1 Ab (Green Peak)

    See Abreview

  • 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.

    See Abreview

  • IRAK-1 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.

References

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) »
See all 9 Publications for this product

Customer reviews and Q&As

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1-6 of 6 Abreviews

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunocytochemistry/ Immunofluorescence
Sample
Rhesus monkey Cell (Colon)
Specification
Colon
Fixative
Paraformaldehyde
Permeabilization
Yes - 0.1% Triton-x-100
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 25°C

Abcam user community

Verified customer

Submitted Feb 21 2013

Abreviews
Application
Western blot
Sample
Mouse Cell lysate - whole cell (RAW macrophages cell line)
Loading amount
50 µg
Specification
RAW macrophages cell line
Treatment
LPS and peptidoglycan
Gel Running Conditions
Reduced Denaturing
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C

Dr. Varkha Agrawal

Verified customer

Submitted Sep 11 2012

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunocytochemistry/ Immunofluorescence
Sample
Mouse Cultured Cells (RAW 264.7)
Specification
RAW 264.7
Fixative
Paraformaldehyde
Permeabilization
Yes - 0.1% Triton-X100 in 2% BSA for 15min
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 2% · Temperature: 22°C

Dr. Mahesh Shivananjappa

Verified customer

Submitted Jun 22 2012

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Human Cell lysate - whole cell (Platelets)
Loading amount
25 µg
Specification
Platelets
Treatment
ADP for 30 min
Gel Running Conditions
Reduced Denaturing (8-16% Biorad Criterion Gel with 18 wells)
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 4°C

Dr. Mahesh Shivananjappa

Verified customer

Submitted Dec 12 2011

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Flow Cytometry
Sample
Human Cell (Platelets)
Specification
Platelets
Preparation
Cell harvesting/tissue preparation method: PL were isolated spinning Platelet rich plasma on Histopaque
Sample buffer: PBS
Fixation
Paraformaldehyde
Permeabilization
Yes - 0.1% Triton-X100 in 2% BSA for 15min
Gating Strategy
Platelets

Abcam user community

Verified customer

Submitted Dec 22 2010

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Human Cell lysate - whole cell (PBMC)
Loading amount
25 µg
Specification
PBMC
Treatment
HIV infected
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C

Dr. Karppiah Muthumani

Verified customer

Submitted Jun 27 2007

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

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