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Albumin (ALB) Colorimetric Assay Kit (Bromocresol Green Method)
SKU: E-BC-K057-M-500
Albumin (ALB) Colorimetric Assay Kit, High Sensitivity (Bromocresol Green Method)
This kit has a higher sensitivity (0.08 g/L) than the E-BC-K057-S kit (0.5 g/L).
| SKU # | E-BC-K057-M |
| Detection Instrument | Microplate reader (620-640 nm) |
| Detection Method | Colorimetric method |
Product Details
Properties
| Synonyms | ALB |
| Sample Type | Serum, plasma |
| Sensitivity | 0.08 g/L |
| Detection Range | 0.08-15 g/L |
| Detection Method | Colorimetric method |
| Assay type | Quantitative |
| Assay time | 20 min |
| Precision | Average inter-assay CV: 4.600% | Average intra-assay CV: 1.500% |
| Other instruments required | Micropipettor, Vortex mixer |
| Other reagents required |
Normal saline (0.9% NaCl) |
| Storage | Reagent 1: 2-8℃, Reagent 2: -20℃ |
| Valid period | 12 months |
Images
A A-R Mohamed et al found that selenium nanoparticle can alleviate cardio-hepatic damage in type II diabetes. Albumin (ALB) level in rat serum was determined using ALB colorimetric assay kit (E-BC-K057-M).
Table. Effect of different treatments (metformin at a dose of (500 mg/kg) and /or CTS-Se-NPs (2 mg Se/kg per day) in HFD/STZ diabetic rats on glucose level, insulin, lipid profile and hepatic oxidative stress.
| Control | HFD/STZ | STZ+MET | STZ+CTS- SeNPs |
STZ+MET+CTS -SeNPs |
|
| TP (g/dl) | 7.17±.07 c | 2.85±12 a | 5.31±.07 b | 5.41±.08 b | 6.88±.11 c |
| Albumin (g/dl) | 3.45±.06 c | 1.39±.03 a | 2.59±.11 b | 2.65±.04 b | 3.44±.06 c |
ALB level was significantly induced in STZ treated group.
Dilution of Sample
It is recommended to take 2~3 samples with expected large difference to do pre-experiment before formal experiment and dilute the sample according to the result of the pre-experiment and the detection range (0.08-15 g/L).
The recommended dilution factor for different samples is as follows (for reference only):
| Sample type | Dilution factor |
| Human serum | 8-15 |
| Human plasma | 8-15 |
| HepG2 supernatant | 1 |
| Mouse plasma | 8-15 |
| Rat serum | 8-15 |
Note: The diluent is normal saline (0.9% NaCl) or PBS (0.01 M, pH 7.4).
Detection Principle
Bromocresol green (BCG) is widely used as protein staining agent. BCG can combine the albumin in pH 4.0~4.2 to form an albumin-BCG complex. And the color changed from yellow to green. The depth of color is proportional to the concentration of albumin. The content of albumin in serum can be calculated indirectly by measuring the OD value at 630 nm.
Kit Components & Storage
| Item | Component | Size 1(48 T) | Size 2(96 T) | Storage |
| Reagent 1 | Chromogenic Agent Stock Solution | 3 mL ×1 vial | 6 mL ×1 vial | 2-8℃, 12 months, shading light |
| Reagent 2 | 20 g/L Standard Solution | 1.2 mL ×1 vial | 1.2 mL × 2 vials | -20℃, 12 months |
| 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 human serum 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 (g/L) | 0.65 | 3.50 | 9.60 |
| %CV | 1.8 | 1.4 | 1.3 |
Inter-assay Precision
Three human serum samples were assayed 20 times in duplicate by three operators to determine precision between assays.
| Parameters | Sample 1 | Sample 2 | Sample 3 |
| Mean (g/L) | 0.65 | 3.50 | 9.60 |
| %CV | 4.3 | 4.8 | 4.7 |
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 95%.
| Standard 1 |
Standard 2 |
Standard 3 | |
| Expected Conc. (g/L) | 1.5 | 4 | 10.5 |
| Observed Conc. (g/L) | 1.5 | 3.7 | 9.9 |
| Recovery rate(%) | 98 | 93 | 9.4 |
Sensitivity
The analytical sensitivity of the assay is 0.08 g/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 (g/L) | 0 | 1.0 | 2.0 | 3.5 | 5.0 | 8.0 | 12 | 15 |
| Average OD | 0.122 | 0.170 | 0.227 | 0.309 | 0.391 | 0.552 | 0.740 | 0.871 |
| Absoluted OD | 0 | 0.048 | 0.105 | 0.187 | 0.269 | 0.430 | 0.618 | 0.749 |