Product Pathways - NF-kB Signaling
PathScan® Total IRAK4 Sandwich ELISA Kit #13336
|13336S||1 Kit (96 assays)||---||In Stock||---|
|13336||carrier free and custom formulation / quantity||email request|
When ordering five or more kits, please contact us for processing time and pricing at email@example.com.
Already purchased this product? Write a Review.
|Kit Includes||Volume||Solution Color|
|Total IRAK4 Rabbit Antibody Coated Microwells||96 tests|
|Total IRAK4 Mouse mAb Detection Antibody||11 ml||Green|
|Anti-mouse IgG, HRP-linked Antibody||11 ml||Red|
|TMB Substrate #7004||11 ml||Colorless|
|STOP Solution #7002||11 ml||Colorless|
|Sealing Tape||2 sheets|
|ELISA Wash Buffer (20X)||25 ml||Colorless|
|ELISA Sample Diluent||25 ml||Blue|
|Cell Lysis Buffer (10X) #9803||15 ml||Yellowish|
Note: 12 8-well modules – Each module is designed to break apart for 8 tests.
Storage: Kit should be stored at 4°C with the exception of Cell Lysis Buffer, which is stored at –20°C (packaged separately).
PathScan® Total IRAK4 Sandwich ELISA Kit is a solid phase sandwich enzyme-linked immunosorbent assay (ELISA) that detects endogenous levels of total IRAK4 protein. An IRAK4 specific antibody has been coated onto the microwells. After incubation with cell lysates, IRAK4 protein is captured by the coated antibody. Following extensive washing, an IRAK4 mouse mAb is added to detect the captured IRAK4 protein. Anti-Mouse IgG, HRP-linked antibody is then used to recognize the bound detection antibody. HRP substrate, TMB, is added to develop color. The magnitude of optical density for this developed color is proportional to the quantity of total IRAK4 protein.
Specificity / Sensitivity
PathScan® Total IRAK4 Sandwich ELISA Kit recognizes endogenous levels of IRAK4 protein in human cells, as shown in Figure 1. The kit sensitivity is shown in Figure 2. This kit detects proteins from the indicated species, as determined through in-house testing, but may also detect homologous proteins from other species.
Figure 1: IRAK4 protein from KARPAS-299 cells, serum-starved overnight and either left untreated (-) or treated with Human Interleukin-1β (hIL-1β) #8900 (50 ng/ml, 15 min; +), was detected using the PathScan® Total IRAK4 Sandwich ELISA Kit #13336. The absorbance readings at 450 nm are shown in the top figure while the corresponding western blot using IRAK4 Antibody #4363 is shown in the bottom figure.
Interleukin-1 (IL-1) receptor-associated kinase (IRAK) is a serine/threonine-specific kinase that can be coprecipitated in an IL-1-inducible manner with the IL-1 receptor (1). The mammalian family of IRAK molecules contains four members (IRAK1, IRAK2, IRAK3/IRAK-M, and IRAK4). The binding of IL-1 to IL-1 receptor type I (IL-1RI) initiates the formation of a complex that includes IL-1RI, AcP, MyD88, and IRAKs (2). IRAK undergoes autophosphorylation shortly after IL-1 stimulation. The subsequent events involve IRAK dissociation from the IL-1RI complex, its ubiquitination, and its association with two membrane-bound proteins: TAB2 and TRAF6. The resulting IRAK-TRAF6-TAB2 complex is then released into the cytoplasm where it activates protein kinase cascades, including TAK1, IKKs, and the stress-activated kinases (3).
Have you published research involving the use of our products? If so we'd love to hear about it. Please let us know!
For Research Use Only. Not For Use In Diagnostic Procedures.
PathScan® is a trademark of Cell Signaling Technology, Inc.
Cell Signaling Technology® is a trademark of Cell Signaling Technology, Inc.