Product nameAnti-IRAK-1 antibody [EPR19667]
See all IRAK-1 primary antibodies
DescriptionRabbit monoclonal [EPR19667] to IRAK-1
Tested applicationsSuitable for: WB, ICC/IFmore details
Species reactivityReacts with: Human
Recombinant fragment within Human IRAK-1 aa 200-450. The exact sequence is proprietary.
Database link: P51617
- WB: HeLa, HEK-293, HepG2 and A549 whole cell lysates. ICC/IF: HeLa cells.
Storage instructionsShipped 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 bufferPreservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol, 0.05% BSA
Concentration information loading...
PurityProtein A purified
Our Abpromise guarantee covers the use of ab208009 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. Detects a band of approximately 80 kDa (predicted molecular weight: 76 kDa).|
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
Lane 1: Wild type HAP1 whole cell lysate (40 µg)
Lane 2: IRAK-1 knockout HAP1 whole cell lysate (40 µg)
Lane 3: HeLa whole cell lysate (40 µg)
Lane 4: HEK293 whole cell lysate (40 µg)
Lanes 1 - 4: Merged signal (red and green). Green - ab208009 observed at 85 kDa. Red - loading control, ab8245, observed at 37 kDa.
ab208009 was shown to specifically react with IRAK-1 when IRAK-1 knockout samples were used. Wild-type and IRAK-1 knockout samples were subjected to SDS-PAGE. Ab208009 and ab8245 (Mouse anti GAPDH loading control) were incubated overnight at 4°C at 200 dilution and 1/10000 dilution respectively. Blots were developed with 800CW Goat anti Rabbit and 680CW Goat anti Mouse secondary antibodies at 1/10000 dilution for 1 hour at room temperature before imaging.
All lanes : Anti-IRAK-1 antibody [EPR19667] (ab208009) at 1/1000 dilution
Lane 1 : HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate
Lane 2 : HEK-293 (Human epithelial cell line from embryonic kidney) whole cell lysate
Lane 3 : HepG2 (Human liver hepatocellular carcinoma cell line) whole cell lysate
Lane 4 : A549 (Human lung carcinoma cell line) whole cell lysate
Lysates/proteins at 10 µg per lane.
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution
Predicted band size: 76 kDa
Observed band size: 68,80 kDa why is the actual band size different from the predicted?
Blocking/Dilution buffer: 5% NFDM/TBST.
Exposure time: Lane 1/3/4: 1 minute; Lane 2: 30 seconds.
There are two splice variants for IRAK-1, IRAK-1b and IRAK-1c,respectively. The product recognizes full length IRAK-1 (80 kDa) and IRAK-1c (68 kDa) (PMID: 16690127).
Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HeLa (Human epithelial cell line from cervix adenocarcinoma) cells labeling IRAK-1 with ab208009 at 1/100 dilution, followed by Goat Anti-Rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing mainly cytoplasmic staining on HeLa cell line. The nuclear counter stain is DAPI (blue).
The negative controls are as follows:
-ve control 1: ab208009 at 1/100 dilution, followed by Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) (ab150120) secondary at 1/1000 dilution.