产品编号 ybEB409Hu 适用生物 Homo sapiens (Human,人)
血红蛋白(HB)检测试剂盒
检测范围 1.23-100μg/mL 灵敏度 0.57μg/mL
样本类型 Serum, plasma, tissue homogenates, erythrocyte lysates, cell culture supernates and other biological fluids.
实验时长 2.5h 实验方法 竞争抑制法
规格 96T
ELISA Kit for Hemoglobin (HB)
FOR IN VITRO AND RESEARCH USE ONLY, NOT FOR USE IN CLINICAL DIAGNOSTIC PROCEDURES!
Organism species |
Homo sapiens (Human) |
Product No. |
CEB409Hu |
Sample type |
Serum, plasma, tissue homogenates, erythrocyte lysates, cell culture supernates and other biological fluids. |
Format |
96T |
Assay length |
2.5 hours |
Detection range |
1.23-100μg/mL The standard curve concentrations used for the ELISA’s were 100μg/mL, 33.33μg/mL, 11.11μg/mL, 3.7μg/mL, 1.23μg/mL |
Sensitivity |
The minimum detectable dose of this kit is typically less than 0.57μg/mL. |
Specificity
This assay has high sensitivity and excellent specificity for detection of Hemoglobin (HB).
No significant cross-reactivity or interference between Hemoglobin (HB) and analogues was observed.
Recovery
血红蛋白(HB)检测试剂盒 Matrices listed below were spiked with certain level of recombinant Hemoglobin (HB) and the recovery rates were calculated by comparing the measured value to the expected amount of Hemoglobin (HB) in samples.
Matrix |
Recovery range (%) |
Average(%) |
serum(n=5) |
96-103 |
101 |
EDTA plasma(n=5) |
79-105 |
81 |
Precision
Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Hemoglobin (HB) were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level Hemoglobin (HB) were tested on 3 different plates, 8 replicates in each plate.
CV(%) = SD/meanX100
Intra-Assay: CV<10%
Inter-Assay: CV<12%
Linearity
The linearity of the kit was assayed by testing samples spiked with appropriate concentration of Hemoglobin (HB) and their serial dilutions. The results were demonstrated by the percentage of calculated concentration to the expected.
Sample |
1:2 |
1:4 |
1:8 |
1:16 |
serum(n=5) |
94-102% |
95-103% |
80-92% |
86-103% |
EDTA plasma(n=5) |
96-105% |
79-105% |
92-101% |
88-99% |
Stability
血红蛋白(HB)检测试剂盒 The stability of kit is determined by the loss rate of activity. The loss rate of this kit is less than 5% within the expiration date under appropriate storage condition.
To minimize extra influence on the performance, operation procedures and lab conditions, especially room temperature, air humidity, incubator temperature should be strictly controlled. It is also strongly suggested that the whole assay is performed by the same operator from the beginning to the end.
Reagents and materials provided
Reagents |
Quantity |
Reagents |
Quantity |
Pre-coated, ready to use 96-well strip plate |
1 |
Plate sealer for 96 wells |
4 |
Standard |
2 |
Standard Diluent |
1×20mL |
Detection Reagent A |
1×120µL |
Assay Diluent A |
1×12mL |
Detection Reagent B |
1×120µL |
Assay Diluent B |
1×12mL |
TMB Substrate |
1×9mL |
Stop Solution |
1×6mL |
Wash Buffer (30 × concentrate) |
1×20mL |
Instruction manual |
1 |
Assay procedure summary
1. Prepare all reagents, samples and standards;
2. Add 50µL standard or sample to each well.
And then add 50µL prepared Detection Reagent A immediately.
Shake and mix. Incubate 1 hour at 37oC;
3. Aspirate and wash 3 times;
4. Add 100µL prepared Detection Reagent B. Incubate 30 minutes at 37oC;
5. Aspirate and wash 5 times;
6. Add 90µL Substrate Solution. Incubate 15-25 minutes at 37oC;
7. Add 50µL Stop Solution. Read at 450 nm immediately.
Test principle
血红蛋白(HB)检测试剂盒 This assay employs the competitive inhibition enzyme immunoassay technique. A monoclonal antibody specific to Hemoglobin (HB) has been pre-coated onto a microplate. A competitive inhibition reaction is launched between biotin labeled Hemoglobin (HB) and unlabeled Hemoglobin (HB) (Standards or samples) with the pre-coated antibody specific to Hemoglobin (HB). After incubation the unbound conjugate is washed off. Next, avidin conjugated to Horseradish Peroxidase (HRP) is added to each microplate well and incubated. The amount of bound HRP conjugate is reverse proportional to the concentration of Hemoglobin (HB) in the sample. After addition of the substrate solution, the intensity of color developed is reverse proportional to the concentration of Hemoglobin (HB) in the sample.
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