Human Immunology ELISA Kits 1

Human IkB alpha (Inhibitory Subunit of NF Kappa B Alpha) CLIA Kit (HUES01254)

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HUES01254

Description

ELISA Kit Technical ManualMSDS

Human IkB alpha (Inhibitory Subunit of NF Kappa B Alpha) CLIA Kit

This kit uses Sandwich-CLIA as the method. The micro CLIA plate provided in this kit has been pre-coated with an antibody specific to Human IkB alpha . Standards or samples are added to the appropriate micro CLIA plate wells and combined with the specific antibody. Then a biotinylated detection antibody specific for Human IkB alpha and Avidin-Horseradish Peroxidase (HRP) conjugate are added to each micro plate well successively and incubated. Free components are washed away. The substrate solution is added to each well. Only those wells that contain Human IkB alpha, biotinylated detection antibody and Avidin-HRP conjugate will appear fluorescence. The Relative light unit (RLU) value is measured spectrophotometrically by the Chemiluminescence immunoassay analyzer. The RLU value is positively associated with the concentration of Human IkB alpha. The concentration of Human IkB alpha in the samples can be calculated by comparing the RLU of the samples to the standard curve.

Human IkB alpha (Inhibitory Subunit of NF Kappa B Alpha) CLIA Kit Information

Target Name:IkB alpha
Target Synonym:
UNIProt ID:P25963
Target Species:Human
Size:96T
Type:Sandwich
Detection Type:CLIA
Detection Range:31.25--2000 pg/mL
Sensitivity:18.75 pg/mL
Tested Sample Types:Serum, Plasma, Cell supernatant

Human IkB alpha (Inhibitory Subunit of NF Kappa B Alpha) CLIA Kit (HUES01254) Assay procedure

    1. Set standard, test sample and control (zero) wells on the pre-coated plate and record theirpositions. It is recommended to measure each standard and sample in duplicate. Note: addall solutions to the bottom of the plate wells while avoiding contact with the well walls. Ensuresolutions do not foam when adding to the wells.
    2. Aliquot 100 µL of standard solutions into the standard wells.
    3. Add 100 µL of Sample / Standard dilution buffer into the control (zero) well.
    4. Add 100 µL of properly diluted sample (serum, plasma, tissue homogenates and otherbiological fluids.) into test sample wells.
    5. Cover the plate with the sealer provided in the kit and incubate for 90 min at 37 °C.
    6. Aspirate the liquid from each well, do not wash. Immediately add 100 µL of BiotinylatedDetection Ab working solution to each well. Cover the plate with a plate seal and gently mix.Incubate for 1 hour at 37 °C.
    7. Aspirate or decant the solution from the plate and add 350 µL of wash buffer to each welland incubate for 1-2 minutes at room temperature. Aspirate the solution from each well andclap the plate on absorbent filter paper to dry. Repeat this process 3 times. Note: a microplatewasher can be used in this step and other wash steps.
    8. Add 100 µL of HRP Conjugate working solution to each well. Cover with a plate seal andincubate for 30 min at 37 °C.
    9. Aspirate or decant the solution from each well. Repeat the wash process for five times asconducted in step 7.
    10. Add 100 µL of Substrate mixture solution to each well. Cover with a new plate seal andincubate for no more than 5 min at 37 °C. Protect the plate from light.
    11. Determine the RLU value of each well immediately.

Human IkB alpha (Inhibitory Subunit of NF Kappa B Alpha) CLIA Kit (HUES01254) Protein Information

UniProt Protein Function:Inhibits the activity of dimeric NF-kappa-B/REL complexes by trapping REL dimers in the cytoplasm through masking of their nuclear localization signals. On cellular stimulation by immune and proinflammatory responses, becomes phosphorylated promoting ubiquitination and degradation, enabling the dimeric RELA to translocate to the nucleus and activate transcription.
NCBI Summary:This gene encodes a member of the NF-kappa-B inhibitor family, which contain multiple ankrin repeat domains. The encoded protein interacts with REL dimers to inhibit NF-kappa-B/REL complexes which are involved in inflammatory responses. The encoded protein moves between the cytoplasm and the nucleus via a nuclear localization signal and CRM1-mediated nuclear export. Mutations in this gene have been found in ectodermal dysplasia anhidrotic with T-cell immunodeficiency autosomal dominant disease. [provided by RefSeq, Aug 2011]
UniProt Code:P25963
NCBI GenInfo Identifier:126682
NCBI Gene ID:4792
NCBI Accession:P25963.1
UniProt Secondary Accession:P25963,B2R8L6,
UniProt Related Accession:P25963
Molecular Weight:35,609 Da
NCBI Full Name:NF-kappa-B inhibitor alpha
NCBI Synonym Full Names:NFKB inhibitor alpha
NCBI Official Symbol:NFKBIA
NCBI Official Synonym Symbols:IKBA; MAD-3; NFKBI
NCBI Protein Information:NF-kappa-B inhibitor alpha
UniProt Protein Name:NF-kappa-B inhibitor alpha
UniProt Synonym Protein Names:I-kappa-B-alpha; IkB-alpha; IkappaBalpha; Major histocompatibility complex enhancer-binding protein MAD3
Protein Family:NF-kappa-B inhibitor
UniProt Gene Name:NFKBIA
UniProt Entry Name:IKBA_HUMAN
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Additional Information

Product Type:
ELISA Kit
ELISA Type:
CLIA Kit
Size:
96 Assays
Sensitivity:
18.75pg/mL
Range:
31.25-2000pg/mL
ELISA Type:
Sandwich
Reactivity:
Human
Sample Type:
Serum, plasma and other biological fluids
Research Area:
Immunology
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