ELISA Kit for Hemoglobin (HB) Cavia (Guinea pig ) Sandwich ELISA

Hgb; Haemoglobin; Heterotetramer(αβ)2

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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
  • SEB409Gu.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) 87-95 90
EDTA plasma(n=5) 92-99 95

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) 98-105% 78-101% 82-96% 90-104%
EDTA plasma(n=5) 97-105% 86-101% 78-98% 89-103%

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 1×120µL 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 100µL standard or sample to each well. Incubate 1 hours at 37°C;
3. Aspirate and add 100µL prepared Detection Reagent A. Incubate 1 hour at 37°C;
4. Aspirate and wash 3 times;
5. Add 100µL prepared Detection Reagent B. Incubate 30 minutes at 37°C;
6. Aspirate and wash 5 times;
7. Add 90µL Substrate Solution. Incubate 10-20 minutes at 37°C;
8. Add 50µL Stop Solution. Read at 450nm immediately.

ELISA Kit for Hemoglobin (HB)

Test principle

The test principle applied in this kit is Sandwich enzyme immunoassay. The microtiter plate provided in this kit has been pre-coated with an antibody specific to Hemoglobin (HB). Standards or samples are then added to the appropriate microtiter plate wells with a biotin-conjugated antibody specific to Hemoglobin (HB). Next, Avidin conjugated to Horseradish Peroxidase (HRP) is added to each microplate well and incubated. After TMB substrate solution is added, only those wells that contain Hemoglobin (HB), biotin-conjugated antibody and enzyme-conjugated Avidin will exhibit a change in color. The enzyme-substrate reaction is terminated by the addition of sulphuric acid solution and the color change is measured spectrophotometrically at a wavelength of 450nm ± 10nm. The concentration of Hemoglobin (HB) in the samples is then determined by comparing the O.D. of the samples to the standard curve.

Citations

  • Effects of ectoine on behavioural, physiological and biochemical parameters of Daphnia magnaPubmed:25460046
  • Comparative Proteomics Identifies Host Immune System Proteins Affected by Infection with Mycobacterium bovisPubmed:27027307
  • The intracellular bacterium Anaplasma phagocytophilum selectively manipulates the levels of vertebrate host proteins in the tick vector Ixodes scapularis.pubmed:27561965
  • Elevated levels of protein AMBP in cerebrospinal fluid of women with preeclampsia compared to normotensive pregnant women.pubmed:27615121
  • Chicory inulin ameliorates type 2 diabetes mellitus and suppresses JNK and MAPK pathways in vivo and in vitropubmed:28105758
  • Oxidized Hemoglobin Is Antigenic and Immunogenic in Lupuspubmed:28694810
  • 新規人工酸素運搬体HemoAct を用いた脳虚血再灌流障害治療開発に関する研究:
  • Mural cell-derived laminin-α5 plays a detrimental role in ischemic strokePubmed: 30777135
  • Decylubiquinone suppresses breast cancer growth and metastasis by inhibiting angiogenesis via the ROS/p53/BAI1 signaling pathwayPubmed: 32020421
  • Water-extractable zinc protoporphyrin IX in Parma ham predominantly exists as complexes with hemoglobin and myoglobin
  • Effects of Human Serum Albumin on the Fluorescence Intensity and Tumor Imaging Properties of IR‐780 Dye34687567

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