SAA ELISA Kit, Human

Catalog number: KHA0011C

Novex™  Related applications: Protein Assays and Analysis

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The Human Serum Amyloid A (SAA) ELISA research-use-only kit is to be used for the quantitative determination of serum amyloid A in samples (see sample types indicated) using 96-well plates and a microplate reader. The assay will recognize both natural and recombinant forms of this target.

Performance characteristics

• Sensitivity: <4 ng/mL
• Standard curve range: 9.4–600 ng/mL
• Sample type(s): serum, plasma, cell culture supernatant
• Specificity: natural and recombinant human SAA (serum amyloid A)
• Cross-reactivity: see kit manual for cross-species and/or cross-target reactivity
• Sample volume: 100 μL
• Total assay time: 3 hours

Rigorous validation

Each manufactured lot of this ELISA kit is quality tested for criteria such as sensitivity, specificity, precision, and lot-to-lot consistency. See manual for more information on validation.

Principle of the method

The human SAA solid-phase sandwich ELISA (enzyme-linked immunosorbent assay) is designed to measure the amount of the target bound between a matched antibody pair. A target-specific antibody has been pre-coated in the wells of the supplied microplate. Samples, standards, or controls are then added into these wells and bind to the immobilized (capture) antibody. The sandwich is formed by the addition of the second (detector) antibody, binding to the target on a different epitope from the capture antibody. A conjugated enzyme has been incorporated into the assay. After incubation and washing steps to rid the microplate of unbound substances, a substrate solution is added that reacts with the enzyme-antibody-target complex to produce measurable signal. The intensity of this signal is directly proportional to the concentration of target present in the original specimen.

Target information

Serum amyloid A (SAA) proteins comprise a family of small (12–14 kDa, 104–112 amino acid residues), differentially expressed proteins that are highly conserved among vertebrates. SAA proteins are involved in the acute phase responses; these are the immediate early host responses to inflammation. During the acute phase, circulating SAA levels are increased by 100–1,000 fold, reaching concentrations of up to one milligram per milliliter. The human SAA gene family maps to chromosome 11. In humans, four SAA genes and three protein products have been identified: human SAA1 and SAA2 are designated the acute phase SAA (A-SAA) isoforms; while, SAA4 is constitutively expressed, and SAA3 is a pseudogene. Mouse SAA gene family maps to chromosome 7. In mouse, five SAA genes and four protein products have been identified: mouse SAA1, SAA2, SAA3 are the acute phase isoforms, SAA5 is constitutively expressed, and SAA4 is a pseudogene. A-SAAs belong to a category of acute phase proteins that also includes C-reactive protein (CRP), haptoglobin, ceruloplasmin, complement components C3 and C4, a1-acid glycoprotein, a1-proteinase inhibitor, and fibrinogen. In humans, both CRP and SAA are synthesized during the acute phase; whereas, in mouse, SAA is the predominant acute phase protein.

These cytokines, in turn, signal the stromal cells to express secondary inflammatory mediators, IL-6, IL-8, and monocyte chemoattractant protein. Leukocytes attracted to the area produce additional cytokines. The cytokines produced at the site of injury, in particular IL-1 and IL-6, enter the plasma and provide stimuli for the liver to produce A-SAAs. Other cytokines that have been implicated in the regulation of SAA synthesis include LIF, CNTF, oncostatin M, cardiotropin, and possibly NGF. Analysis of SAA promoter regions reveals control by several transcription factors, including NFκB, C/EBP, YY1, SAF/Sp1, and AP-2. AP-2 acts to repress transcription in non-liver tissues. NFκB and C/EBP act synergistically to enhance transcription, while YY1 antagonizes the function of NFκB and, therefore, serves to silence transcription.

Upon synthesis, SAA is released into the bloodstream where it immediately binds the HDL particles. Binding to HDL is known to protect the SAAs from degradation by proteolytic enzymes. There are a number of important homeostatic functions associated with the circulating SAA-HDL complexes; these functions have been categorized as immune modulation, lipid transport, and anti-inflammatory. The circulating SAA is known to act as a chemoattractant and recruit additional monocytes, leukocytes, mast cells, and T lymphocytes to the site of inflammation. It also aides the tissue regeneration process by activating matrix metalloproteinases such as collagenase and strolemysin. Binding of SAA to HDL alters the reverse cholesterol transport function of HDL, allowing for the delivery of cholesterol for the site of repair. The SAA-HDL complex also serves to remove excess cholesterol released by the damaged tissue. The anti-inflammatory activities of circulating SAA include inhibition of lymphocyte cell function, inhibition of TNF-α- and IL-1-induced fevers, and inhibition of platelet aggregation. SAA is also known to bind neutrophils, abrogate the oxidative burst response, and prevent oxidative tissue damage.

Chronically elevated SAA levels are implicated in a group of protein misfolding disorders known as the amyloid A amyloidoses. These include reactive amyloidosis, caused by chronic inflammation or recurrent acute inflammation, familial Mediterranean fever, systemic AA amyloidosis, and visceral AA amyloidosis. Amyloid A amyloidoses are characterized by the deposition of insoluble plaques composed principally of proteolytically cleaved A-SAA. These plaques are primarily deposited in organ sites of inflammation and ultimately result in the degeneration of the affected organ. Despite the potential for causing fatal amyloidoses, SAAs are believed to play a protective role for the host. This hypothesis is supported by the observations that there is immediate, robust induction of SAAs in response to inflammatory signals and that SAAs are highly conserved through evolution.

Related links

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For Research Use Only. Not for use in diagnostic procedures.


Gene Aliases: SAA, SAA4
Shipping Condition: Wet Ice
Marker: SAA (Serum Amyloid A)
Gene ID: 6287
Species: Human
Gene Symbol: SAA@
Sensitivity: 4 ng/mL
Target Gene: serum amyloid A1 cluster
Label or Dye: HRP (Horseradish Peroxidase)
Product Size: 480 assays
Concentration: 9.4-600 ng⁄ml
Sample Volume: 10 µl
Incubation Time: 3 hrs
Detection Method: Colorimetric
Research Category: Immunology, Inflammation
Sample Type (Specific): Plasma, Serum
For Use With (Equipment): Microplate Reader

Contents & storage

Pre-coated plate(s), standard or calibrator, detector antibody, HRP conjugate, diluents, wash buffer, substrate(s), and stop solution. Store kit at 2-8°C. See product manual for detailed contents and storage conditions for maximum stability.


Manuals & protocols