• Antigen :
• NF-κB p65

• Alternative Names :
• NFKB3 ; p65 ; RelA

• Background :
• This kit uses Sandwich-ELISA as the method. The micro ELISA plate provided in this kit has been pre-coated with an antibody specific to Human IgE .Standards or samples are added to the appropriate micro ELISA plate wells and combined with the specific antibody. Then a biotinylated detection antibody specific for Human IgE 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 IgE ,biotinylated detection antibody and Avidin-HRP conjugate will appear fluorescence. The OD value is measured spectrophotometrically at a wavelength of 450nm. The OD value is positively associated with the concentration of Human IgE . The concentration of Human IgE in the samples can be calculated by comparing the OD of the samples to the standard curve.

• Description :
• 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.
  

  Concentration(ng/mL)	O.D	Average	Corrected
  20	2.274 2.304	2.289	2.219
  10	1.624 1.658	1.641	1.571
  5	0.877 0.869	0.873	0.803
  2.5	0.431 0.459	0.445	0.375
  1.25	0.238 0.22	0.229	0.159
  0.63	0.178 0.16	0.169	0.099
  0.31	0.119 0.121	0.12	0.05
  0	0.063 0.077	0.07000000000000001	--

• Reactivity :
• Mouse

• Specificity :
• This kit recognizes Mouse NF-κB p65 in samples. No significant cross-reactivity or interference between Mouse NF-κB p65 and analogues was observed.

• Assay Time :
• 4.5h

• Components :
• 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.
  

  Item	Specifications	Storage
  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

• Detection Method :
• Colormetric

• Detection Range :
• 0.313--20ng/mL

• Linearity :
• Samples were spiked with high concentrations of Mouse NF-κB p65 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)
  1:2	Range (%)	91-104	98-110	93-107
  	Average (%)	97	104	101
  1:4	Range (%)	88-102	86-98	83-97
  	Average (%)	93	92	88
  1:8	Range (%)	88-104	85-97	86-98
  	Average (%)	96	90	91
  1:16	Range (%)	90-102	83-93	84-96
  	Average (%)	96	88	89

• Method Type :
• Sandwich

• Precision :
• Intra-assay Precision (Precision within an assay): 3 samples with low, mid range and high level Mouse NF-κB p65 were tested 20 times on one plate, respectively.
  Inter-assay Precision (Precision between assays): 3 samples with low, mid range and high level Mouse NF-κB p65 were tested on 3 different plates, 20 replicates in each plate.
  

  	Intra-assay Precision			Inter-assay Precision
  Sample	1	2	3	1	2	3
  n	20	20	20	20	20	20
  Mean (ng/mL)	1.05	1.94	6.71	1.15	2.11	6.27
  Standard	deviation	0.05	0.10	0.30	0.06	0.12	0.22
  C V (%)	4.76	5.15	4.47	5.22	5.69	3.51

• Recovery :
• The recovery of Mouse NF-κB p65 spiked at three different levels in samples throughout the range of the assay was evaluated in various matrices.
  

  Sample Type	Range (%)	Average Recovery (%)
  Serum (n=5)	92-107	98
  EDTA plasma (n=5)	87-101	94
  Cell culture media (n=5)	88-99	94

• Sample Type :
• Serum, Plasma, Cell supernatant

• Sensitivity :
• 0.19 ng/mL

• Manual :
•