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This ELISA kit applies to the in vitro quantitative determination of Human NF-κBp65 concentrations in serum, plasma and other biological fluids.
This ELISA kit uses the Sandwich-ELISA principle. The micro ELISA plate provided in this kit has been pre-coated with an antibody specific to Human NF-κBp65. Standards or samples are added to the micro ELISA plate wells and combined with the specific antibody. Then a biotinylated detection antibody specific for Human NF-κBp65 and Avidin-Horseradish Peroxidase (HRP) conjugate are added successively to each micro plate well and incubated. Free components are washed away. The substrate solution is added to each well. Only those wells that contain Human NF-κBp65, biotinylated detection antibody and Avidin-HRP conjugate will appear blue in color. The enzyme-substrate reaction is terminated by the addition of stop solution and the color turns yellow. The optical density (OD) is measured spectrophotometrically at a wavelength of 450 nm ± 2 nm. The OD value is proportional to the concentration of Human NF-κBp65. You can calculate the concentration of Human NF-κBp65 in the samples by comparing the OD of the samples to the standard curve.
|Detection Range||0.16—10 ng/mL|
|Sample Volume Required Per Well||100μL|
|Sample Type||Serum, plasma and other biological fluids|
This kit recognizes Human NF-κBp65 in samples. No significant cross-reactivity or interference between Human NF-κBp65 and analogues was observed.
As the OD values of the standard curve may vary according to the conditions of the actual assay performance (e.g. operator, pipetting technique, washing technique or temperature effects), the operator should establish a standard curve for each test. Typical standard curve and data is provided below for reference only.
Intra-assay Precision (Precision within an assay): 3 samples with low, mid range and high level Human NF-κBp65 were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, mid range and high level Human NF-κBp65 were tested on 3 different plates, 20 replicates in each plate.
|Intra-assay Precision||Inter-assay Precision|
|C V (%)||5.56||4.03||5.29||5.77||5.04||3.31|
The recovery of Human NF-κBp65 spiked at three different levels in samples throughout the range of the assay was evaluated in various matrices.
|Sample Type||Range (%)||Average Recovery (%)|
|EDTA plasma (n=5)||92-105||100|
|Cell culture media (n=5)||86-97||92|
Samples were spiked with high concentrations of Human NF-κBp65 and diluted with Reference Standard & Sample Diluent to produce samples with values within the range of the assay.
|Serum (n=5)||EDTA plasma (n=5)||Cell culture media (n=5)|
An unopened kit can be stored at 4℃ for 1 month. If the kit is not used within 1 month, store the items separately according to the following conditions once the kit is received.
|Micro ELISA Plate(Dismountable)||8 wells ×12 strips||-20℃, 6 months|
|Reference Standard||2 vials|
|Concentrated Biotinylated Detection Ab (100×)||1 vial, 120 μL|
|Concentrated HRP Conjugate (100×)||1 vial, 120 μL||-20℃(shading light), 6 months|
|Reference Standard & Sample Diluent||1 vial, 20 mL||4℃, 6 months|
|Biotinylated Detection Ab Diluent||1 vial, 14 mL|
|HRP Conjugate Diluent||1 vial, 14 mL|
|Concentrated Wash Buffer (25×)||1 vial, 30 mL|
|Substrate Reagent||1 vial, 10 mL||4℃(shading light)|
|Stop Solution||1 vial, 10 mL||4℃|
|Plate Sealer||5 pieces|
|Product Description||1 copy|
|Certificate of Analysis||1 copy|
Note: All reagent bottle caps must be tightened to prevent evaporation and microbial pollution.
The volume of reagents in partial shipments is a little more than the volume marked on the label, please use accurate measuring equipment instead of directly pouring into the vial(s).
1.Add 100 μL standard or sample to each well. Incubate for 90 min at 37℃.
2.Remove the liquid.
3.Add 100 μL Biotinylated Detection Ab. Incubate for 1 hour at 37℃. Aspirate and wash 3 times.
4.Add 100 μL HRP Conjugate. Incubate for 30 min at 37℃. Aspirate and wash 5 times.
5.Add 90 μL of Substrate Reagent. Incubate for 15 min at 37℃.
6.Add 50 μL Stop Solution.
7.Read at 450 nm immediately. Calculation of results.
D******************eSubmitted [ Jan 20 2019 ]
Asked: Dear colleagues, We would like to offer Human NF-κB p65(Nuclear Factor Kappa B p65) ELISA Kit. How can we do it?
adminSubmitted [ Jan 20 2019 ]
Answered: Dear Dr Natalia Chebotare, Thanks for your interest in our product? This is Cathy Mao for your service. Can you advise where are you from? The catalogue number for the kit inquired is E-EL-H1388, retail price is $580, usually we will charge $50 per order as freight cost which will be under our DHL or FedEx account, if you have your own express, this cost will be free of charge for you. Lead time is 3 working days and delivery will be made upon receipt of payment or bank slip, shelf life of our kit is 6 months. If you have any other question, please feel free to ask me. Thanks and have a nice day~ Regards, Cathy