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ELISA Kit for Hemoglobin (HB) Homo sapiens (Human) Competition ELISA

Hgb; Haemoglobin; Heterotetramer(αβ)2

  • Product No.CEB409Hu
  • Organism SpeciesHomo sapiens (Human) Same name, Different species.
  • Sample Typeserum, plasma, tissue homogenates, erythrocyte lysates, cell culture supernates and other biological fluids
  • Test MethodCompetitive Inhibition
  • Assay Length2h
  • Detection Range6.17-500ug/mL
  • SensitivityThe minimum detectable dose of this kit is typically less than 2.62ug/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.
  • ApplicationsEnzyme-linked immunosorbent assay for Antigen Detection.
  • DownloadInstruction Manual
  • UOM 48T96T 96T*5 96T*10 96T*100 In stock
  • FOB US$ 279.00 US$ 398.00 US$ 1,791.00 US$ 3,383.00 US$ 27,860.00
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  • ELISA Kit for Hemoglobin (HB) Packages (Simulation)
  • ELISA Kit for Hemoglobin (HB) Packages (Simulation)
  • ELISA Kit for Hemoglobin (HB) Results demonstration
  • CEB409Hu.jpg Typical Standard Curve
  • Certificate ISO9001: 2008, ISO13485: 2003 Registered

Recovery

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

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 Assay Diluent A 1×12mL
Detection Reagent B 1×120µL Assay Diluent B 1×12mL
Reagent Diluent 1×300µL Stop Solution 1×6mL
TMB Substrate 1×9mL Instruction manual 1
Wash Buffer (30 × concentrate) 1×20mL

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 37°C;
3. Aspirate and wash 3 times;
4. Add 100µL prepared Detection Reagent B. Incubate 30 minutes at 37°C;
5. Aspirate and wash 5 times;
6. Add 90µL Substrate Solution. Incubate 10-20 minutes at 37°C;
7. Add 50µL Stop Solution. Read at 450 nm immediately.

ELISA Kit for Hemoglobin (HB)

Test principle

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.

Citations

  • Comparative Biochemistry and Physiology Part C: Toxicology &amp; PharmacologyEffects of ectoine on behavioural, physiological and biochemical parameters of Daphnia magnaPubmed:25460046
  • PLoS One.Comparative Proteomics Identifies Host Immune System Proteins Affected by Infection with Mycobacterium bovisPubmed:27027307
  • Parasites & VectorsThe intracellular bacterium Anaplasma phagocytophilum selectively manipulates the levels of vertebrate host proteins in the tick vector Ixodes scapularis.pubmed:27561965

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