货号：17-486

品牌：Merck Millipore

规格：

目录价：￥6845.00

市场价格：￥5818.25

会员价格：￥5476.00

金山科研平台，产品价格货期咨询微信：jinshanbio Description: phospho-IκB (Ser32) STAR ELISA Kit View All» Trade Name: Upstate (Millipore) View All» Qty/Pk: 96 assays View All» Background Information: I. TEST PRINCIPLE: The colorimetric STAR (Signal Transduction Assay Reaction) ELISA kit is a solid phase sandwich enzyme linked immunosorbent assay that provides a fast, sensitive method to detect specific levels of signaling targets in whole cell extracts. The I_κB plate is coated with a specific mouse monoclonal I_κB capture antibody on the microwells of the 96-well clear plate. Sample lysate or the standard included in the kit are incubated in the microwells allowing I_κB antigen to be captured in the plate wells. The plate is then washed to remove any un-bound nonspecific material. A specific rabbit anti-phospho-I_κB antibody is added to detect the captured I_κB on the plate well that is phosphorylated on Ser32. The unbound detection antibody is washed away followed by incubation with an HRP-conjugated anti-rabbit antibody. This allows for a sensitive enzymatic detection of the sample. After the addition of TMB substrate and stop solution the absorbance is measured at 450 nm using a plate reader. The entire assay takes less than 5 hours to complete with minimal hands-on time. Many of the reagents are supplied in ready-to use formulations for ease of use. The kit also includes a standard that is run as both a positive control and to generate a standard curve for I_κB measurement. II. BACKGROUND: NF_κB signaling is a critical regulator not only of immune function, but also of proliferation versus apoptosis in response to various stimuli. In a majority of unstimulated cells, the NF_κB transcription factors exist in their inactive form and are retained in the cytoplasm by the bound inhibitory I_κB proteins that prevent it from entering the nucleus, thus keeping NF_κB in an inactive state. Upon stimulation by multiple inducers including viruses or cytokines, such as TNFα, IL-1, or PMA, I_κBα is rapidly phosphorylated by IKK (I_κB Kinase), which phosphorylates I_κB on serines 32 and 36, allowing recognition by the ubiquitin ligase machinery, leading to its polyubiquination and degradation, resulting in the release of the NF_κB complex. Once I_κB is degraded, NF_κB is able to initiate transcription. The NF_κB complex then translocates to the nucleus where it activates gene transcription. NF_κB induces the transcription of its own inhibitor, I_κB_α, causing an autoregulatory mechanism of NF_κB activity and generating the inactive form of NF_κB. The newly formed nuclear NF_κB- I_κB_α complexes are then exported out to the cytoplasm, thereby reestablishing the cytoplasmic pool of inactive NF_κB complexes primed for another round of activation to take place. The wide variety of genes regulated by NF_κB includes those encoding cytokines, chemokines, adhesion molecules, acute phase proteins, and inducible effector enzymes. Mutant I_κB in which serines 32 and 36 are changed to alanines, is not phosphorylated, and therefore not degraded. Cells expressing this protein are not able to activate NF_κB, providing a useful tool to study the role of NF_κB in various pathways and processes. View All» Key Applications: ELISA View All» Application Notes: • All kit reagents should be brought to room temperature (20°C to 25°C) prior to use. • Do not use reagents beyond the expiration date of the kit. • Do not mix or interchange reagent from various kit lots. • Manual Plate Washing: Vigorous washing and complete removal of all liquid by aspiration at the end of each washing step is very important to obtain low background values. Gentle agitation during the wash steps or a 2-3 minute soak may reduce background values. • The desiccant enclosed in the 96-well capture plate pouch will keep the plate stable when stored at 2° to 8°C should the pouch loose its seal during shipping. View All» Species Reactivity: Human View All» Usage Statement: Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals. View All» Entrez Gene Summary: NFKB1 (MIM 164011) or NFKB2 (MIM 164012) is bound to REL (MIM 164910), RELA (MIM 164014), or RELB (MIM 604758) to form the NFKB complex. The NFKB complex is inhibited by I-kappa-B proteins (NFKBIA or NFKBIB, MIM 604495), which inactivate NF-kappa-B by trapping it in the cytoplasm. Phosphorylation of serine residues on the I-kappa-B proteins by kinases (IKBKA, MIM 600664, or IKBKB, MIM 603258) marks them for destruction via the ubiquitination pathway, thereby allowing activation of the NF-kappa-B complex. Activated NFKB complex translocates into the nucleus and binds DNA at kappa-B-binding motifs such as 5-prime GGGRNNYYCC 3-prime or 5-prime HGGARNYYCC 3-prime (where H is A, C, or T; R is an A or G purine; and Y is a C or T pyrimidine).[supplied by OMIM] View All» UniProt Summary: FUNCTION: SwissProt: P25963 # Inhibits NF-kappa-B by complexing with and trapping it in the cytoplasm. May be involved in regulation of transcriptional responses to NF-kappa-B, including cell adhesion, immune and proinflammatory responses, apoptosis, differentiation and growth. Controlled by sequential serine-phosphorylation, ubiquitination and degradation. Tyrosine-phosphorylation could only lead to dissociation from NF-kappa-B.SIZE: 317 amino acids; 35609 Da SUBUNIT: Interacts with p65 (RELA). Interacts with NKIRAS1 and NKIRAS2. Interacts with HBV protein X.SUBCELLULAR LOCATION: Cytoplasm.PTM: Phosphorylated; disables inhibition of NF-kappa-B DNA-binding activity. & Ubiquitinated; subsequent to stimulus-dependent phosphorylation on serines, polyubiquitination targets the protein for rapid degradation via the ubiquitin system.DISEASE: SwissProt: P25963 # Defects in NFKBIA are a cause of autosomal dominant anhidrotic ectodermal dysplasia with immunodeficiency (AD-EDA-ID) [MIM:164008]. Ectodermal dysplasias (EDs) constitute a heterogeneous group of developmental disorders affecting tissues of ectodermal origin. EDs are characterized by abnormal development of two or more ectodermal structures such as hair, teeth, nails and sweat glands, with or without any additional clinical sign. Each combination of clinical features represents a different type of ectodermal dysplasia.SIMILARITY: SwissProt: P25963 ## Belongs to the NF-kappa-B inhibitor family. & Contains 5 ANK repeats. View All» Brand Family: Upstate View All» Sensitivity: Sensitivity: 1 Unit/mL. Range of Detection: 1.6 to 100 Units/mL View All» Storage Conditions: 1 year at 4°C from date of shipment View All» Modifications: Phosphorylation View All» Gene Symbol:

• NFKBIA

• IKB-alpha

• IKBA

• I-kappa-B-alpha

• MAD-3

• MAD3

• IkappaBalpha

• NFKBI
  View All» Product Name: phospho-IκB (Ser32) STAR ELISA Kit View All» Materials Required but Not Delivered: 1. Multi-channel or repeating pipettes 2. Plate shaker (optional) 3. Pipettors & tips capable of accurately measuring 1-1000 μL 4. Graduated serological pipettes 5. 96-well microtiter Plate Reader with 450 nm filter 6. Graphing software for plotting data or graph paper for manual plotting of data 7. Microfuge tubes for standard and sample dilutions 8. Mechanical vortex 9. 1 liter container 10. Distilled or deionized water View All» Analytes Available: IκB View All» Quality Assurance: Routinely evaluated by ELISA. View All» Components:
  • 1. Capture Plate pre-coated with anti-I_κB antibody: (Part No. 17-486A) One pre-coated 96-stripwell immunoplate sealed in a foil pouch.

  • 2. Anti-phospho- I_κB (ser32) detection antibody: (Part No. 17-486B) One bottle (6 mL) of anti- phospho-I_κB (Ser32) detection antibody containing sodium azide, ready to use.

  • 3. ELISA Diluent: (Part No. 17-486C) One bottle (25 mL) of ELISA Diluent containing sodium azide, ready to use.

  • 4. 25X ELISA Wash Buffer: (Part No. 17-486D) One bottle (50 mL) of 25X ELISA Wash Buffer.

  • 5. Anti-Rabbit IgG HRP conjugate: (Part No. 17-486E) One vial (125 μL) of 100X anti-rabbit HRP conjugate.

  • 6. HRP Diluent: (Part No. 17-486F) One bottle (25 mL) of HRP Diluent.

  • 7. TMB Solution: (Part No. 17-486G) One bottle (25 mL) of stabilized tetramethylbenzidine (TMB), ready to use.

  • 8. Stop Solution: (Part No. 17-486H) One bottle (25 mL) of stop solution, ready to use.

  • 9. Phosphorylated I_κB (Ser32) Standard: (Part No. 17-486I) Two vials of phosphorylated I_κB standard (Ser32), lyophilized.

  • 10. Plate Covers: Two plate covers.
    View All» UniProt Number: P25963 View All» Detection Methods: Colorimetric View All» Entrez Gene Number: NM_020529.1 View All»
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