Cell Ferrous Iron Colorimetric Assay Kit

Cell Ferrous Iron Colorimetric Assay Kit

SKU #	E-BC-K881-M
Detection Instrument	Microplate reader (590-600 nm, optimum wavelength: 593 nm)
Detection method	Colorimetric method

Product Details

Properties

Sample type	Cells
Sensitivity	0.4 μmol/L
Detection range	0.4-50 μmol/L
Detection method	Colorimetric method
Assay type	Quantitative
Assay time	40 min
Precision	Average inter-assay CV: 1.500% | Average intra-assay CV: 1.300%
Storage	2-8℃
Valid period	12 months

 

Images

Y Zhang et al investigate the ferroptosis of renal tubular epithelial cells that promoting chronic renal allograft interstitial fibrosis. Cell ferrous ion level in HK-2 cell were determined using cell ferrous ion colorimetric assay kit (E-BC-K881-M).

Intracellular ferrous iron was significantly increased in cells treated with TNF-α. (***P<0.0001)

 

Dilution of Sample

The recommended dilution factor for different samples is as follows (for reference only):

Sample type	Dilution factor
HepG2 Cell	1
molt-4 Cell	1
Jurkat Cell	1
HEL Cell	1

Note: The diluent is buffer solution. For the dilution of other sample types, please do pretest to confirm the dilution factor

 

Detection Principle

Iron is one of the metal elements in organism and has important physiological functions. Ferrous ion is a key element in heme and hemoglobin and plays an important role in many biochemical reactions. Ferrous ions (Fe2+) in samples can bind with probe to form complexes, which has a maximum absorption peak at 593 nm. The concentration of ferrous ions can be calculated by measuring the OD value at 593 nm indirectly.

Kit Components & Storage

Item	Component	Size 1(48 T)	Size 2(96 T)	Storage
Reagent 1	Buffer Solution	45 mL × 1 vial	35 mL × 2 vials	2-8℃, 12 month shading light
Reagent 2	Control Solution	5 mL × 1 vial	10 mL × 1 vial	2-8℃, 12 month shading light
Reagent 3	Chromogenic Solution	5 mL × 1 vial	10 mL × 1 vial	2-8℃, 12 month shading light
Reagent 4	10 mmol/L Iron Standard	2 mL ×1 vial	2 mL ×1 vial	2-8℃, 12 month shading light
Reagent 5	Standard Protectant	Powder × 1 vial	Powder × 1 vial	2-8℃, 12 month shading light
	Microplate	96 wells		No requirement
	Plate Sealer	2 pieces

 

Note: The reagents must be stored strictly according to the preservation conditions in the above table. The reagents in different kits cannot be mixed with each other. For a small volume of reagents, please centrifuge before use, so as not to obtain sufficient amount of reagents.

 

Technical Data:

Parameter:

Intra-assay Precision

Three HepG2 cell samples were assayed in replicates of 20 to determine precision within an assay (CV = Coefficient of Variation).

Parameters	Sample 1	Sample 2	Sample 3
Mean (μmol/L)	2.80	162.00	46.00
%CV	1.5	1.3	1.1

Inter-assay Precision

Three HepG2 cell samples were assayed 20 times in duplicate by three operators to determine precision between assays.

Parameters	Sample 1	Sample 2	Sample 3
Mean (μmol/L)	2.80	162.00	46.00
%CV	1.2	1.6	1.7

Recovery

Take three samples of high concentration, middle concentration and low concentration to test the samples of each concentration for 6 times parallelly to get the average recovery rate of 99%.

	Standard 1	Standard 2	Standard 3
Expected Conc. (μmol/L)	6.8	17.5	36.5
Observed Conc. (μmol/L)	6.9	17.2	35.4
Recovery rate(%)	102	98	97

 

Sensitivity

The analytical sensitivity of the assay is 0.4 μmol/L. This was determined by adding two standard deviations to the mean O.D. obtained when the zero standard was assayed 20 times, and calculating the corresponding concentration.

 

Standard Curve

As the OD value of the standard curve may vary according to the conditions of the actual assay performance (e.g. operator, pipetting technique or temperature effects), so the standard curve and data are provided as below for reference only:

Concentration (μmol/L)	0	5	10	15	20	30	40	50
Average OD	0.045	0.079	0.110	0.136	0.163	0.221	0.288	0.352
Absoluted OD	0.000	0.035	0.065	0.091	0.118	0.177	0.244	0.307