Total Iron Ion Assay Kit (for Biochemical Assay Kit)
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Categories : 1. Chemical and Reagents , Biochemicals , ELISA Kits & Assay Kits , Servicebio ,
Brand : Servicebio
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Product Information
Product Name | Cat. No. | Spec. |
Total Iron Ion Assay Kit | G4301-48T | 48 T |
G4301-96T | 96 T |
Product Description/Introduction
Iron is one of the important metallic elements in living organisms and is involved in various physiological activities such as hematopoiesis, oxygen transport, enhancement of immunity, and synthesis and reactivity of certain enzymes. With the introduction of the concept of iron death in recent years, the change of iron ion content as one of the characteristics of iron death also indicates that the process of iron ion metabolism has an important relationship with the health and disease of organisms.
This product is Total Iron Assay Kit. The detection principle is that under special conditions, iron ions are fully released from the sample and quickly reduced to ferrous ions by the reducing agent in the reagent. Then ferrous ions combine with the detection probe to produce a colored substance, and within a certain concentration range, the color of the resulting substance is proportional to the concentration of ferrous ions.
Storage and Shipping Conditions
Ship with wet ice; Store at 2-8 away from light for 6 months.
Product Content
Component Number | Component | G4301-48T | G4301-96T |
G4301-1 | Iron ion standard (1 mmol/L) | 0.5 mL | 0.5 mL |
G4301-2 | Total Iron Detection Buffer | 20 mL | 40 mL |
G4301-3 | Total Iron Detection Probe | 5 mL | 10 mL |
Note: Product specifications are based on the tissue and cell sample testing assay system.
Assay Protocol / Procedures
1. Sample Preparation:
1.1. Plasma and serum samples: Use the total iron assay buffer in the kit to mix thoroughly at a ratio of 1 to 4 between sample and buffer (if there is precipitation 10,000 g centrifugation and then take the supernatant for detection), for subsequent total iron ion detection;
1.2. Tissue samples: Please use total iron detection buffer for homogenization lysis, in which the ratio of tissue and homogenate is 1 to 9 can be (Example: 30 mg to 270 μL); after homogenization lysis, centrifugation at 4 , 10,000 g for 10-15 min, and the supernatant is taken for the subsequent detection of total iron ions;
1.3. Cell samples: After cell collection, use total iron assay buffer to resuspend the cells, adding 500 μL of homogenate approximately every 1 × 106 cells; after the homogenate is lysed, centrifuge the cells at 4°C, 10,000 g for 10-15 min, and take the supernatant for the subsequent total iron ion assay;
1.4. Iron ion standard: The iron ion standard (1 mmol/L) is diluted in a gradient (e.g., 50/25/12.5/6.25/3.125 μmol/L) using total iron assay buffer and used for data analysis in the standard curve method after subsequent assaying with the samples to be measured; or appropriately diluted to a known concentration and used for data analysis in the standard curve method after subsequent assaying with the samples to be measured. Standard conversion method data analysis;
2. Total Iron Ion Detection:
Note: Plasma and serum samples and tissue and cell samples are subject to different detection systems and procedures, so the reference standards need to be detected under the corresponding systems and procedures before the corresponding analyses can be performed.
2.1. Plasma and serum sample testing - the modus operandi is shown in the table below;
standard tube | blank tube | sample tube | |
Total Iron Ion Standard | 50 μL | ||
Total Iron Detection Buffer | 50 μL | ||
Samples to be tested | 50 μL | ||
Total Iron Detection Buffer | 50 μL | 50 μL | 50 μL |
Mix well and incubate at 37 for 15 min | |||
Detection Probes | 100 μL | 100 μL | 100 μL |
Incubate at 37°C for 40 min and detect the absorbance at 593 nm using an enzyme meter. | |||
2.2. Tissue and Cell Sample Assay - The modus operandi is shown in the table below;
standard tube | blank tube | sample tube | |
Total Iron Ion Standard | 100 μL | ||
Total Iron Detection Buffer | 100 μL | ||
Samples to be tested | 100 μL | ||
Total Iron Detection Buffer | 100 μL | 100 μL | 100 μL |
Mix well and incubate at 37 for 15 min | |||
Detection Probes | 100 μL | 100 μL | 100 μL |
Incubate at 37 for 40 min, centrifuge at 10000 g for 5 min, take 200 μL of supernatant in an enzyme plate, and detect the absorbance at 593 nm by enzyme marker. | |||
3. Data analysis:
3.1. Before data analysis, the protein concentration of the tissue or cell samples to be tested needs to be calculated. The protein concentration of the sample to be tested can be determined with the BCA Protein Quantification Kit (G2026).
3.2. Standardized Curve Method:
3.2.1. Draw a standard curve based on the standard group-blank group value data of gradient dilution: Y=aX+b(Y is the standard group-blank group value; X is the standard concentration of iron ion; a is the slope of the standard curve;b is the intercept of the standard curve);
3.2.2. Calculation of total iron ion concentration in serum plasma samples
Total ferrous ion concentration per unit sample (μmoL/L) = [(Sample group value - Blank group value) - b]/a × sample dilution
3.2.3. Calculation of total iron ion concentration in tissue and cell samples:
Total ferrous ion concentration per unit sample (μmol/gprot) = [(Sample group value - Blank group value) - b]/a × sample dilution / sample protein concentration (gprot/L)
3.3. Standard product conversion method:
3.3.1. Calculation of MDA concentration per unit serum plasma sample:
Total iron ion concentration per unit sample (μmol/L) = (sample group value - blank group value)/(standard group value - blank group value) × Total iron ion concentration of standard product (μmol/L) × sample dilution
3.3.2. MDA concentration calculation of tissue and cell samples:
Unit sample total iron ion concentration (μmol/gprot) = (sample group value - blank group value)/(standard group value - blank group value) × standard total iron ion concentration (μmol/L) × sample dilution ratio/tissue or cell protein concentration (gprot/L)
Note
1. Gently shake the reagents before use to ensure that the components are homogeneous.
2. When analyzing the data by the standard conversion method, the concentration of the sample to be tested and the standard should be within the linear detection range of the kit; when analyzing the data by the standard curve method, make sure that the R2 of the standard curve is more than 0.99.
3. Avoid handling or transferring samples with ferrous objects.
4. For your health and safety, please wear lab coat and gloves during operation.
Appendix: Technical parameters
Testing Indicators | Total Iron Ion Concentration |
Sample type | Serum (plasma), tissue, cells |
Detection sensitivity and range | 0.762 μmol/L；0.762-50 μmol/L |
Recovery rate | >99% |
Intra-batch CV | <5% |
Inter-batch CV | <5% |
For Research Use Only!
