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Invitrogen
Human IkB alpha (Total/Phospho) InstantOne™ ELISA Kit
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FIGURE: 1 / 3
Human IkB alpha (Total/Phospho) InstantOne™ ELISA Kit
Representative data for IkB alpha (Total/Phospho) Human InstantOne™ ELISA. HeLa cells were unstimulated (-) or stimulated with 10 ng/mL TNF alpha for 5 min (+). Cells were analyzed for total (left) or phosphorylated (right) IkBa by InstantOne™ ELISA.
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Product Specifications
Sample type/volume
Cell Lysate
20 µL
Hands-on time
1 hr 20 min
Time-to-result
1 hr 10 min
Homogenous (no wash)
No
Instrument
Absorbance Microplate Reader
Product size
96 Tests
Contents
Capture Antibody
Detection Antibody Reagent
InstantOne™ Assay Plate
Cell Lysis Buffer
Enhancer Solution
Wash Buffer (10X)
Substrate Reagent (TMB)
Stop Solution
Positive Control Cell Lysate (lyophilized)
Plate seal
Detection Antibody Reagent
InstantOne™ Assay Plate
Cell Lysis Buffer
Enhancer Solution
Wash Buffer (10X)
Substrate Reagent (TMB)
Stop Solution
Positive Control Cell Lysate (lyophilized)
Plate seal
Shipping conditions
Wet ice
Storage
2-8°C
Protein name
IkB alpha
Target site-specificity
Target sub-specificity
Total
Assay kit format
InstantOne ELISA
Detector antibody conjugate
Biotin
Label or dye
HRP
About This Kit
The InstantOne™ ELISA is specifically engineered for accurate measurement of total and phosphorylated human IkBa in cell lysates. Detection of mouse and rat IkBa is expected. The InstantOne™ ELISA kit allows for fast analysis of samples in approximately one hour. All reagents used in a traditional sandwich ELISA are added in solution to a plate followed by a wash step and detection with the TMB colorimetric substrate.
Target information
IkB-alpha is a 40 kDa protein that functions to inhibit NF- kappaB activity. The inhibition occurs via protein-protein interaction between I kappaB proteins and NF- kappaB dimers in the cytosol. The interaction of I kappa B-alpha with NF- kappaB masks the nuclear localization sequence of NF- kappaB, preventing NF- kappaB translocation to the nucleus. A variety of stimuli can activate gene expression by liberating NF- kappaB through the degradation of I kappaB alpha. These stimuli include the proinflammatory cytokines TNF- alpha and IL-1 beta, chemokines, PMA, growth factors, LPS, UV irradiation, viral infection, as well as various chemical and physical stresses. In humans, the gene is located on the q arm of chromosome 14. Activation of NFkB requires that IkB be phosphorylated on specific serine residues, which results in targeted degradation of IkB. IkB kinase alpha (IKK alpha), previously designated CHUK, interacts with IkB-alpha and specifically phosphorylates IkB-alpha on the sites that trigger its degradation Serines 32 and 36. IKK alpha appears to be critical for NFkB activation in response to proinflammatory cytokines. Phosphorylation of IkB by IKK alpha is stimulated by the NFkB inducing kinase (NIK), which itself is a central regulator for NFkB activation in response to TNF and IL-1. The functional IKK complex contains three subunits, IKK alpha, IKK beta and IKK gamma, and each appear to make essential contributions to IkB phosphorylation.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.
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