ELISA Kit for Serum Amyloid A (SAA) Homo sapiens (Human) Competition ELISA

SA-A; SAA1; PIG4; TP53I4; Amyloid protein A; Amyloid fibril protein AA; Serum amyloid A-1 protein

  • ELISA Kit for Serum Amyloid A (SAA) Packages (Simulation)
  • ELISA Kit for Serum Amyloid A (SAA) Packages (Simulation)
  • ELISA Kit for Serum Amyloid A (SAA) Results demonstration
  • CEA885Hu.jpg Typical Standard Curve
  • Certificate ISO9001: 2008, ISO13485: 2003 Registered

Recovery

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

Matrix Recovery range (%) Average(%)
serum(n=5) 80-97 82
EDTA plasma(n=5) 91-98 94
heparin plasma(n=5) 91-98 95

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Serum Amyloid A (SAA) were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level Serum Amyloid A (SAA) 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 Serum Amyloid A (SAA) 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) 84-103% 80-95% 85-101% 79-90%
EDTA plasma(n=5) 82-89% 91-99% 96-104% 82-92%
heparin plasma(n=5) 80-101% 89-96% 83-104% 83-94%

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 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 Serum Amyloid A (SAA)

Test principle

This assay employs the competitive inhibition enzyme immunoassay technique. A monoclonal antibody specific to Serum Amyloid A (SAA) has been pre-coated onto a microplate. A competitive inhibition reaction is launched between biotin labeled Serum Amyloid A (SAA) and unlabeled Serum Amyloid A (SAA) (Standards or samples) with the pre-coated antibody specific to Serum Amyloid A (SAA). 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 Serum Amyloid A (SAA) in the sample. After addition of the substrate solution, the intensity of color developed is reverse proportional to the concentration of Serum Amyloid A (SAA) in the sample.

Reference

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  • BMC Complementary and Alternative MedicineIdentification of plasma protein markers common to patients with malignant tumour and Abnormal Savda in Uighur medicine: a prospective clinical studyPubmed:25652121
  • BOSCAEFFECT OF UTERINE IMMUNOMODULATION ON SERUM AMYLOID-A CONCENTRATION AND CONCEPTION RATE IN CYCLIC NON BREEDING COWSTcrjournals:Source
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  • EndocrineRelation of thyroid hormone abnormalities with subclinical inflammatory activity in patients with type 1 and type 2 diabetes mellitusPubMed: 26049370
  • J Assist Reprod Genet.The effect of serum and follicular fluid amyloid-associated protein levels on in vitro fertilization outcome in patients with polycystic ovary syndromePubMed: 26463878
  • International Journal of Clinical and Experimental MedicineCirculating serum amyloid A: a potential biomarker for infection after aneurysmal subarachnoid hemorrhagefiles:ijcem0021201
  • PLoS One.An Anti-β-Amyloid Vaccine for Treating Cognitive Deficits in a Mouse Model of Down SyndromePubmed:27023444
  • United States Patent ApplicationPATHWAY SPECIFIC MARKERS FOR DIAGNOSING IRRITABLE BOWEL SYNDROMEy2016:0139148.html

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