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ELISA Kit for Haptoglobin (Hpt) Homo sapiens (Human) Sandwich ELISA

HP; Hp2-Alpha; Alpha-2-Macroglobulin; Zonulin

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  • ELISA Kit for Haptoglobin (Hpt) Packages (Simulation)
  • ELISA Kit for Haptoglobin (Hpt) Packages (Simulation)
  • ELISA Kit for Haptoglobin (Hpt) Results demonstration
  • SEA817Hu.jpg Typical Standard Curve
  • Certificate ISO9001: 2008, ISO13485: 2003 Registered

Recovery

Matrices listed below were spiked with certain level of recombinant Haptoglobin (Hpt) and the recovery rates were calculated by comparing the measured value to the expected amount of Haptoglobin (Hpt) in samples.

Matrix Recovery range (%) Average(%)
serum(n=5) 87-94 91
EDTA plasma(n=5) 86-101 97
heparin plasma(n=5) 91-99 95

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Haptoglobin (Hpt) were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level Haptoglobin (Hpt) 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 Haptoglobin (Hpt) 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) 79-95% 82-97% 94-101% 93-105%
EDTA plasma(n=5) 91-98% 83-94% 80-101% 86-93%
heparin plasma(n=5) 82-94% 88-97% 82-91% 83-97%

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 Haptoglobin (Hpt)

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 Haptoglobin (Hpt). Standards or samples are then added to the appropriate microtiter plate wells with a biotin-conjugated antibody specific to Haptoglobin (Hpt). 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 Haptoglobin (Hpt), 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 Haptoglobin (Hpt) in the samples is then determined by comparing the O.D. of the samples to the standard curve.

Citations

  • Circulation levels of acute phase proteins in patients with Takayasu arteritisPubMed: 20100644
  • Quantitative trait loci mapping of the mouse plasma proteome (pQTL).NCBI: PMC3567747
  • Docosahexaenoic and eicosapentaenoic acid supplementation does not exacerbate oxidative stress or intravascular haemolysis in homozygous sickle cell patients.Pubmed: 24095588
  • Gut Microbial Dysbiosis May Predict Diarrhea and Fatigue in Patients Undergoing Pelvic Cancer Radiotherapy: A Pilot StudyPubMed: 25955845
  • Application of a new procedure for liquid chromatography/mass spectrometry profiling of plasma amino acid-related metabolites and untargeted shotgun proteomics to identify mechanisms and biomarkers of calcific aortic stenosisS0021967317311858
  • High complement levels in astrocyte‐derived exosomes of Alzheimer diseasePubmed:29406582
  • Effects of Hippophae rhamnoides L. Leaf and Marc Extract with Reduced Tannin Concentration on the Health and Growth Parameters of Newborn Calves
  • Using ruminally protected and nonprotected active dried yeast as alternatives to antibiotics in finishing beef steers: growth performance, carcass traits, blood …Pubmed: 30184125
  • Urinary Proteomics for the Early Diagnosis of Diabetic Nephropathy in Taiwanese PatientsPubmed: 30486327
  • Generation of Pigs Resistant to Highly Pathogenic-Porcine Reproductive and Respiratory Syndrome Virus through Gene Editing of CD163
  • Prolyl endopeptidase-degraded low immunoreactive wheat flour attenuates immune responses in Caco-2 intestinal cells and gluten-sensitized BALB/c micePubmed: 31082461
  • Ruminally protected and unprotected Saccharomyces cerevisiae fermentation products as alternatives to antibiotics in finishing beef steersPubmed: 31410465
  • Deletion of CD163 Exon 7 Confers Resistance to Highly Pathogenic Porcine Reproductive and Respiratory Viruses on Pigs
  • Intestinal permeability and small intestine bacterial overgrowth in excess weight adolescentsPubmed: 33089672
  • Processing Index of Barley Grain and Dietary Undigested Neutral Detergent Fiber Concentration Affected Chewing Behavior, Ruminal pH and Total Tract Nutrient?¡­33474564
  • Influence of niacin application on inflammatory parameters, non-esterified fatty acids and functional status of liver in cows during early lactation
  • Could serum zonulin be an intestinal permeability marker in diabetes kidney disease?34170951
  • The Effect of Feed Supplementation with EM Bokashi® Multimicrobial Probiotic Preparation on Selected Parameters of Sow Colostrum and Milk as Indicators …34596883

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