Human SAA (Serum Amyloid A) CLIA Kit (HUES01152)
- Product Type:
- CLIA Kit
- 96 Assays
- ELISA Type:
- Tested Sample Types:
- Serum, plasma and other biological fluids
|Detection range:||0.31-20 ng/mL|
|Sample type:||Serum, plasma and other biological fluids|
|Repeatability:||CV < 15%|
|Specificity:||This kit recognizes Human SAA in samples. No significant cross-reactivity or interference between Human SAA and analogues was observed.|
This kit uses Sandwich-CLIA as the method. The micro CLIA plate provided in this kit has been pre-coated with an antibody specific to Human SAA. Standards or samples are added to the appropriate micro CLIA plate wells and combined with the specific antibody. Then a biotinylated detection antibody specific for Human SAA and Avidin-Horseradish Peroxidase (HRP) conjugate are added to each micro plate well successively and incubated. Free components are washed away. The substrate solution is added to each well. Only those wells that contain Human SAA, biotinylated detection antibody and Avidin-HRP conjugate will appear fluorescence. The Relative light unit (RLU) value is measured spectrophotometrically by the Chemiluminescence immunoassay analyzer. The RLU value is positively associated with the concentration of Human SAA. The concentration of Human SAA in the samples can be calculated by comparing the RLU of the samples to the standard curve.
|UniProt Protein Function:||SAA1: Major acute phase reactant. Apolipoprotein of the HDL complex. Reactive, secondary amyloidosis is characterized by the extracellular accumulation in various tissues of the SAA1 protein. These deposits are highly insoluble and resistant to proteolysis; they disrupt tissue structure and compromise function. Elevated serum SAA1 protein levels may be associated with lung cancer. Belongs to the SAA family.|
|UniProt Protein Details:|
Protein type:Secreted, signal peptide; Secreted
Chromosomal Location of Human Ortholog: 11p15. 1
Cellular Component: extracellular region
Molecular Function:heparin binding; G-protein-coupled receptor binding
Biological Process: positive regulation of interleukin-1 secretion; receptor-mediated endocytosis; platelet activation; neutrophil chemotaxis; elevation of cytosolic calcium ion concentration; positive regulation of cell adhesion; regulation of protein secretion; negative regulation of inflammatory response; acute-phase response; innate immune response; lymphocyte chemotaxis; macrophage chemotaxis; positive regulation of cytokine secretion
|NCBI Summary:||This gene encodes a member of the serum amyloid A family of apolipoproteins. The encoded protein is a major acute phase protein that is highly expressed in response to inflammation and tissue injury. This protein also plays an important role in HDL metabolism and cholesterol homeostasis. High levels of this protein are associated with chronic inflammatory diseases including atherosclerosis, rheumatoid arthritis, Alzheimer's disease and Crohn's disease. This protein may also be a potential biomarker for certain tumors. Alternate splicing results in multiple transcript variants that encode the same protein. A pseudogene of this gene is found on chromosome 11. [provided by RefSeq, Jun 2012]|
|NCBI GenInfo Identifier:||395406826|
|NCBI Gene ID:||6288|
|NCBI Accession:||P0DJI8. 1|
|UniProt Secondary Accession:||P0DJI8,P02735, P02736, P02737, Q16730, Q16834, Q16835 Q16879, Q3KRB3, Q6FG67, Q96QN0, Q9UCK9,|
|UniProt Related Accession:||P0DJI8|
|NCBI Full Name:||Serum amyloid A-1 protein|
|NCBI Synonym Full Names:||serum amyloid A1|
|NCBI Official Symbol:||SAA1|
|NCBI Official Synonym Symbols:||SAA; PIG4; SAA2; TP53I4|
|NCBI Protein Information:||serum amyloid A-1 protein; serum amyloid A protein; tumor protein p53 inducible protein 4|
|UniProt Protein Name:||Serum amyloid A-1 protein|
|UniProt Synonym Protein Names:||Amyloid fibril protein AA|
|UniProt Gene Name:||SAA1|
|UniProt Entry Name:||SAA1_HUMAN|
As the RLU values of the standard curve may vary according to the conditions of the actual assay performance (e. g. operator, pipetting technique, washing technique or temperature effects), the operator should establish a standard curve for each test. Typical standard curve and data is provided below for reference only.
Intra-assay Precision (Precision within an assay): 3 samples with low, mid range and high level Human SAA were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, mid range and high level Human SAA were tested on 3 different plates, 20 replicates in each plate.
|Intra-assay Precision||Inter-assay Precision|
|C V (%)||8.41||9.72||8.40||10.43||11.92||7.77|
The recovery of Human SAA spiked at three different levels in samples throughout the range of the assay was evaluated in various matrices.
|Sample Type||Range (%)||Average Recovery (%)|
|EDTA plasma (n=5)||86-97||91|
|Cell culture media (n=5)||89-100||94|
Samples were spiked with high concentrations of Human SAA and diluted with Reference Standard & Sample Diluent to produce samples with values within the range of the assay.
|Serum (n=5)||EDTA plasma (n=5)||Cell culture media (n=5)|
An unopened kit can be stored at 4°C for 1 month. If the kit is not used within 1 month, store the items separately according to the following conditions once the kit is received.
|Micro CLIA Plate(Dismountable)||8 wells ×12 strips||-20°C, 6 months|
|Reference Standard||2 vials|
|Concentrated Biotinylated Detection Ab (100×)||1 vial, 120 µL|
|Concentrated HRP Conjugate (100×)||1 vial, 120 µL||-20°C(shading light), 6 months|
|Reference Standard & Sample Diluent||1 vial, 20 mL||4°C, 6 months|
|Biotinylated Detection Ab Diluent||1 vial, 14 mL|
|HRP Conjugate Diluent||1 vial, 14 mL|
|Concentrated Wash Buffer (25×)||1 vial, 30 mL|
|Substrate Reagent A||1 vial, 5 mL||4°C (shading light)|
|Substrate Reagent B||1 vial, 5 mL||4°C (shading light)|
|Plate Sealer||5 pieces|
|Product Description||1 copy|
|Certificate of Analysis||1 copy|
- Set standard, test sample and control (zero) wells on the pre-coated plate and record theirpositions. It is recommended to measure each standard and sample in duplicate. Note: addall solutions to the bottom of the plate wells while avoiding contact with the well walls. Ensuresolutions do not foam when adding to the wells.
- Aliquot 100 µL of standard solutions into the standard wells.
- Add 100 µL of Sample / Standard dilution buffer into the control (zero) well.
- Add 100 µL of properly diluted sample (serum, plasma, tissue homogenates and otherbiological fluids. ) into test sample wells.
- Cover the plate with the sealer provided in the kit and incubate for 90 min at 37 °C.
- Aspirate the liquid from each well, do not wash. Immediately add 100 µL of BiotinylatedDetection Ab working solution to each well. Cover the plate with a plate seal and gently mix. Incubate for 1 hour at 37 °C.
- Aspirate or decant the solution from the plate and add 350 µL of wash buffer to each welland incubate for 1-2 minutes at room temperature. Aspirate the solution from each well andclap the plate on absorbent filter paper to dry. Repeat this process 3 times. Note: a microplatewasher can be used in this step and other wash steps.
- Add 100 µL of HRP Conjugate working solution to each well. Cover with a plate seal andincubate for 30 min at 37 °C.
- Aspirate or decant the solution from each well. Repeat the wash process for five times asconducted in step 7.
- Add 100 µL of Substrate mixture solution to each well. Cover with a new plate seal andincubate for no more than 5 min at 37 °C. Protect the plate from light.
- Determine the RLU value of each well immediately.