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Synthetic peptide, corresponding to C terminal amino acids 700-712 of Human IRAK
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 IRAK1 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.|
ab238 staining IRAK1 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 IRAK1 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 IRAK1 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: IRAK1
Immunofluorescence of IRAK1 in HeLa cells using ab238 at 20 ug/ml.
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