Product datasheet
Anti-IRAK (phospho T387) antibody ab139739
3 Images
Overview
Product name
Anti-IRAK (phospho T387) antibody
Description
Rabbit polyclonal to IRAK (phospho T387)
Tested applications
ELISA, WB
Species reactivity
Reacts with: Human
Predicted to work with: Chimpanzee, Gorilla, Orangutan
Immunogen
Synthetic peptide corresponding to Human IRAK aa 350-450 (phospho T387) conjugated to
Keyhole Limpet Haemocyanin (KLH).
Database link: P51617
Positive control
This antibody gave a positive signal in THP1 whole cell lysate.
Properties
Form
Liquid
Storage instructions
Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C or 80°C. Avoid freeze / thaw cycle.
Storage buffer
pH: 7.40
Preservative: 0.02% Sodium azide
Constituent: PBS
Batches of this product that have a concentration < 1mg/ml may have BSA added as a
stabilising agent. If you would like information about the formulation of a specific lot, please
contact our scientific support team who will be happy to help.
Purity
Immunogen affinity purified
Clonality
Polyclonal
Isotype
IgG
Applications
Our Abpromise guarantee covers the use of ab139739 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Application
ELISA
Abreviews
Notes
Use a concentration of 1 µg/ml.
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Application
Abreviews
WB
Notes
Use a concentration of 1 µg/ml. Detects a band of approximately 74 kDa
(predicted molecular weight: 76 kDa).
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'.
Anti-IRAK (phospho T387) antibody images
Serially diluted ab139739 was bound to
immobilised phospho (P; ab174118) - or
control (C; ab174119) peptides (1 microgram
x mL-1). The antibody was detected by HRPlabelled goat anti-rabbit IgG (ab97080;
diluted 50000 times) and signal was
developed with TMB substrate.
ELISA - Anti-IRAK (phospho T387) antibody
(ab139739)
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All lanes : Anti-IRAK (phospho T387)
antibody (ab139739) at 1 µg/ml
Lane 1 : THP1 (Human acute monocytic
leukemia cell line) Whole Cell Lysate
Lane 2 : THP1 (Human acute monocytic
leukemia cell line) Whole Cell Lysate with
Human IRAK (phospho T387) peptide
(ab174118) at 1 µg/ml
Lane 3 : THP1 (Human acute monocytic
leukemia cell line) Whole Cell Lysate with
Western blot - Anti-IRAK (phospho T387) antibody
Human IRAK peptide (ab174119) at 1 µg/ml
(ab139739)
Lysates/proteins at 25 µg per lane.
Secondary
Goat Anti-Rabbit IgG H&L (HRP) (ab97051)
at 1/10000 dilution
developed using the ECL technique
Performed under reducing conditions.
Predicted band size : 76 kDa
Observed band size : 74 kDa
Exposure time : 16 minutes
This blot was produced using a 10% Bis-tris
gel under the MOPS buffer system. The gel
was run at 200V for 50 minutes before being
transferred onto a Nitrocellulose membrane at
30V for 70 minutes. The membrane was then
blocked for an hour using 5% Bovine Serum
Albumin before being incubated with
ab139739 overnight at 4°C. Antibody binding
was detected using an anti-rabbit antibody
conjugated to HRP, and visualised using ECL
development solution.
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ab139739 was tested using an Indirect ELISA
approach. The wells were coated with peptide
(1µg/ml at 100µl/well) overnight at 4°C,
followed by a 5% BSA blocking step for 1
hour at room temperature. The primary Ab
was then added at a dilution range of 10.00025µg/ml (100µl/well) for 1hr at room
temperature. A HRP-conjugated anti-rabbit
IgG (heavy and light chain) was used as a
secondary antibody at 1:20,000 dilution for
1hr at room temperature.
ELISA - Anti-IRAK (phospho T387) antibody
(ab139739)
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