Signaling Nodes Multi-Target BioGene ELISA Kit

Signaling Nodes Multi-Target BioGene ELISA Kit
Item number Size Datasheet Manual SDS Delivery time Quantity Price
S1013-84.1 1 kit - -

4 - 22 business days

952.00€
 
MultiTarget Sandwich ELISA Kit is a solid phase sandwich enzyme-linked immunosorbent assay... more
Product information "Signaling Nodes Multi-Target BioGene ELISA Kit"
MultiTarget Sandwich ELISA Kit is a solid phase sandwich enzyme-linked immunosorbent assay (ELISA) that combines the reagents necessary to detect endogenous levels of Akt1, phospho-Akt1 (Ser473), phospho-MEK1 (Ser217/221), phospho-p38a MAPK (Thr180/Tyr182), phospho-Stat3 (Tyr705) and phospho-NF-kB p65 (Ser536). These molecules represent convergence points and key regulatory proteins in signaling pathways controlling cellular events such as growth and differentiation, energy homeostasis and the response to stress and inflammation. Sixteen tests are provided for each target protein. Specific assay formulations for the indicated target proteins can be found in the datasheets associated with the individual PathScan(R) Sandwich ELISA Kits?. Briefly, a capture antibody+ has been coated onto the microwells. After incubation with cell lysates, the target protein is captured by the coated antibody. Following extensive washing, a detection antibody+ is added to detect the captured target protein. An HRP-linked secondary antibody is then used to recognize the bound detection antibody. HRP substrate, TMB, is added to develop color. The magnitude of absorbance for this developed color is proportional to the quantity of bound target protein. Nodes Multi-Target Sandwich ELISA Ki detects endogenous levels of six proteins: Akt1, phospho-Akt1 (Ser473), phospho-MEK1 (Ser217/221), phospho-p38a MAPK (Thr180/Tyr182), phospho-Stat3 (Tyr705) and phospho-NF-kB p65 (Ser536). Differential phosphorylation of these proteins can be observed over time in response to various growth factor and cytokine treatments. As shown in Figure 1, stimulation of serum starved NIH/3T3 cells with PDGF generates robust phosphorylation of Akt1 at Ser473 and MEK1 at Ser217/221 but relatively weak phosphorylation of Stat3 at Tyr705. Simultaneous treatment of NIH/3T3 cells with TNF-a and IL-1beta results in phosphorylation of Akt1 at Ser473, MEK1 at Ser217/221, p38a MAPK at Thr180/Tyr182 and NF-kB p65 at Ser536 but no activation of Stat3. By comparison, NIH/3T3 cells respond to IL-6 primarily through the activation of Stat3 rather than Akt1, MEK1, p38a MAPK or NF-kB p65. The level of each target protein (phospho and nonphospho) remains unchanged throughout the 80 minute time course as demonstrated by Western analysis. Akt is a protooncogene with a critical regulatory role in diverse cellular processes including growth, survival and the cell cycle. Akt is also a major regulator of insulin signaling and glucose metabolism (1-4). Akt is activated by PI3 kinase signaling and activation loop phosphorylation at Thr308 by PDK1 and by phosphorylation withing the carboxyl terminus at Ser473 by PDK2 (5-7). MEK1 and MEK2 are dual-specificity protein kinases that function in a mitogen activated protein kinase cascade controlling cell growth and differentiation. Activation of MEK1 and MEK2 occurs through phosphorylation of serine 217 and serine 221 by Raf-like molecules. MEK activates p44 and p42 MAP kinase (8-10). p38 MAP kinase (MAPK) participates in a signaling cascade controlling the cellular response to pro-inflammatory cytokines and a variety of cellular stresses. MKK3, MKK6 and SEK (MKK4) activate p38 MAP kinase by phosphorylation at Thr180 and Tyr182 (11-14). The Stat3 transcription factor is an important signaling molecule for many cytokines and growth factor receptors. Stat3 is activated by phosphorylation at Tyr705, which induces dimerization, nuclear translocation and DNA binding (15,16). , Kit Components:, Akt Rabbit Ab Coated Microwells, 16 tests , Akt1 Mouse Detection Ab, 1x1.8ml , Anti-Mouse IgG HRP-Linked Ab, 1x1.8ml, Phospho-Akt (Ser473) Rabbit Ab Coated Microwells, 16 tests , Akt1 Mouse Detection Ab, 1x1.8ml, Anti-Mouse IgG HRP-Linked Ab, 1x1.8ml, MEK1 Mouse Ab Coated Microwells, 16 tests , Phospho-MEK1/2 (Ser217/221) Rabbit Detection Ab, 1x1.8ml, Anti-Rabbit IgG HRP-Linked Ab, 1x1.8ml, p38alpha MAP Kinase Rabbit Ab Coated Microwells, 16 tests , Phospho-p38alpha MAPK (Thr180/Tyr182) Mouse Detection Ab, 1x1.8ml, Anti-Mouse IgG HRP-Linked Ab, 1x1.8ml, Stat3 Rabbit Ab Coated Microwells, 16 tests , Phospho-Stat3 (Tyr705) Mouse Detection Ab, 1.8ml, Anti-Mouse IgG HRP-Linked Ab, 1x1.8ml, NF-kappaB p65 Mouse Ab Coated Microwells, 16 tests , Phospho-NF-kappaB p65 (Ser536) Rabbit Detection Ab, 1x1.8ml, Anti-Rabbit IgG HRP-Linked Ab, 1x1.8ml, TMB Substrate, 1x11ml , STOP Solution, 1x11ml , 20X Wash Buffer, 1x25ml , Sample Diluent, 1x25ml , Cell Lysis Buffer (10X),1x15ml, Storage and Stability: Store components at 4°C. Stable for 6 months. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap.  , Storage and Stability: Store components at 4°C. Stable for 6 months For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap.
Supplier: United States Biological
Supplier-Nr: S1013-84

Properties

Application: ELISA
Format: Solid Phase

Database Information

Handling & Safety

Storage: -20°C
Shipping: +4°C (International: +4°C)
Caution
Our products are for laboratory research use only: Not for administration to humans!
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