Human Calprotectin (S100A8/S100A9) ELISA Kit is a single-wash 90-min Simplestep used to quantify Human Calprotectin (S100A8/S100A9) with a sensitivity of 13.624 pg/ml. The assay uses a simple mix-wash-read protocol with just one incubation and one wash step.
- Colorimetric Sandwich ELISA - 450 nm readout : works on any standard plate reader
- Different formats for different needs: 10x96 plates for bulk orders
- Design your own immunoassay: we also offer the conjugation-ready antibody pair
Images
Key facts
- Detection method
- Colorimetric
- Sample types
- Serum, Citrate plasma, EDTA Plasma, Cell culture supernatant, Urine, Saliva, Milk, Cell Lysate, Tissue Extracts
- Assay type
- Sandwich (quantitative)
- Reactive species
- Human
- Range
- 31.25 - 2000 pg/mL
- Assay time
- 1h 30m
- Sensitivity
- = 13.624 pg/mL
Reactivity data
| Application | Reactivity | Dilution info | Notes |
|---|---|---|---|
Application sELISA | Reactivity Reacts | Dilution info - | Notes - |
Target data
Function
S100A8 is a calcium- and zinc-binding protein which plays a prominent role in the regulation of inflammatory processes and immune response. It can induce neutrophil chemotaxis and adhesion. Predominantly found as calprotectin (S100A8/A9) which has a wide plethora of intra- and extracellular functions. The intracellular functions include: facilitating leukocyte arachidonic acid trafficking and metabolism, modulation of the tubulin-dependent cytoskeleton during migration of phagocytes and activation of the neutrophilic NADPH-oxidase. Participates also in regulatory T-cell differentiation together with CD69 (PubMed:26296369). Activates NADPH-oxidase by facilitating the enzyme complex assembly at the cell membrane, transferring arachidonic acid, an essential cofactor, to the enzyme complex and S100A8 contributes to the enzyme assembly by directly binding to NCF2/P67PHOX. The extracellular functions involve pro-inflammatory, antimicrobial, oxidant-scavenging and apoptosis-inducing activities. Its pro-inflammatory activity includes recruitment of leukocytes, promotion of cytokine and chemokine production, and regulation of leukocyte adhesion and migration. Acts as an alarmin or a danger associated molecular pattern (DAMP) molecule and stimulates innate immune cells via binding to pattern recognition receptors such as Toll-like receptor 4 (TLR4) and receptor for advanced glycation endproducts (AGER). Binding to TLR4 and AGER activates the MAP-kinase and NF-kappa-B signaling pathways resulting in the amplification of the pro-inflammatory cascade. Has antimicrobial activity towards bacteria and fungi and exerts its antimicrobial activity probably via chelation of Zn(2+) which is essential for microbial growth. Can induce cell death via autophagy and apoptosis and this occurs through the cross-talk of mitochondria and lysosomes via reactive oxygen species (ROS) and the process involves BNIP3. Can regulate neutrophil number and apoptosis by an anti-apoptotic effect; regulates cell survival via ITGAM/ITGB and TLR4 and a signaling mechanism involving MEK-ERK. Its role as an oxidant scavenger has a protective role in preventing exaggerated tissue damage by scavenging oxidants. Can act as a potent amplifier of inflammation in autoimmunity as well as in cancer development and tumor spread. The iNOS-S100A8/A9 transnitrosylase complex directs selective inflammatory stimulus-dependent S-nitrosylation of GAPDH and probably multiple targets such as ANXA5, EZR, MSN and VIM by recognizing a [IL]-x-C-x-x-[DE] motif; S100A8 seems to contribute to S-nitrosylation site selectivity. (Microbial infection) Upon infection by human coronavirus SARS-CoV-2, may induce expansion of aberrant immature neutrophils in a TLR4-dependent manner.
Additional Targets
S100A9
Alternative names
CAGA, CFAG, MRP8, S100A8, Protein S100-A8, Calgranulin-A, Calprotectin L1L subunit, Cystic fibrosis antigen, Leukocyte L1 complex light chain, Migration inhibitory factor-related protein 8, S100 calcium-binding protein A8, Urinary stone protein band A, MRP-8, p8
What's included?
Human Calprotectin (S100A8/S100A9) ELISA Kit is a single-wash 90-min Simplestep used to quantify Human Calprotectin (S100A8/S100A9) with a sensitivity of 13.624 pg/ml. The assay uses a simple mix-wash-read protocol with just one incubation and one wash step.
- Colorimetric Sandwich ELISA - 450 nm readout : works on any standard plate reader
- Different formats for different needs: 10x96 plates for bulk orders
- Design your own immunoassay: we also offer the conjugation-ready antibody pair
Key facts
- Detection method
- Colorimetric
- Sample types
- Serum, Citrate plasma, EDTA Plasma, Cell culture supernatant, Urine, Saliva, Milk, Cell Lysate, Tissue Extracts
- Assay type
- Sandwich (quantitative)
- Reactive species
- Human
- Range
- 31.25 - 2000 pg/mL
- Assay time
- 1h 30m
- Assay Platform
- Pre-coated microplate (12 x 8 well strips)
- Sensitivity
- = 13.624 pg/mL
- General recovery
- 107%
Precision
Intra assay
| Sample | n | mean | SD | C.V. |
|---|---|---|---|---|
Sample Plasma | n 8 | mean - | SD - | C.V. 4.5 |
Inter assay
| Sample | n | mean | SD | C.V. |
|---|---|---|---|---|
Sample Plasma | n 3 | mean - | SD - | C.V. 4.4 |
Recovery
Sample specific recovery
| Sample type | Average % | Range |
|---|---|---|
Sample type Serum | Average % = 107 | Range 98 - 112 % |
Sample type Citrate plasma | Average % = 105 | Range 101 - 109 % |
Sample type EDTA Plasma | Average % = 100 | Range 99 - 101 % |
Sample type Cell culture supernatant | Average % = 94 | Range 85 - 101 % |
Sample type Urine | Average % = 105 | Range 105 - 106 % |
Sample type Saliva | Average % = 95 | Range 85 - 105 % |
Sample type Milk | Average % = 113 | Range 104 - 119 % |
Sample type Cell Lysate | Average % = 105 | Range 96 - 110 % |
Sample type Tissue Extracts | Average % = 106 | Range 99 - 113 % |
Storage
- Shipped at conditions
- Blue Ice
- Appropriate short-term storage conditions
- +4°C
- Appropriate long-term storage conditions
- +4°C
- Storage information
- +4°C
Notes
Human Calprotectin (S100A8/S100A9) SimpleStep ELISA® kit is a single-wash 90 min sandwich ELISA designed for the quantitative measurement of Calprotectin (S100A8/S100A9) protein in Serum, Cit plasma, EDTA Plasma, Cell culture supernatant, Urine, Saliva, Milk, and Cell Lysate. Quantitate Human Calprotectin (S100A8/S100A9) with 13.624 pg/ml sensitivity.
SimpleStep ELISA® technology employs capture antibodies conjugated to an affinity tag that is recognized by the monoclonal antibody used to coat our SimpleStep ELISA® plates. This approach to sandwich ELISA allows the formation of the antibody-analyte sandwich complex in a single step, significantly reducing assay time. See the SimpleStep ELISA® protocol summary in the image section for further details. Our SimpleStep ELISA® technology provides several benefits:
-Single-wash protocol reduces assay time to 90 minutes or less
-High sensitivity, specificity and reproducibility from superior antibodies
-Fully validated in biological samples
-96-wells plate breakable into 12 x 8 wells strips, available in 10-pack (10 x 96-well plates)
-Also available in a fully validated 384-well format.
Abcam has not and does not intend to apply for the REACH Authorisation of customers' uses of products that contain European Authorisation list (Annex XIV) substances.
It is the responsibility of our customers to check the necessity of application of REACH Authorisation, and any other relevant authorisations, for their intended uses.
Supplementary info
- This supplementary information is collated from multiple sources and compiled automatically.
- Activity summary
S100A9 and S100A8 also known as calprotectin or MRP8/14 when they form a complex together are small calcium-binding proteins with molecular masses of approximately 13 kDa and 11 kDa respectively. These proteins show high expression levels in neutrophils and monocytes serving important roles in the immune response. Both S100A9 and S100A8 display the ability to bind calcium ions changing their conformation and functional status. This calcium-induced structural change is essential for interacting with other proteins and cellular components.
- Biological function summary
The S100A9 and S100A8 proteins regulate the inflammatory response by modulating immune cell activity. They function as heterodimers forming complexes that influence cell adhesion migration and the production of reactive oxygen species. Such activities are integral to both acute and chronic inflammation. The proteins achieve these effects by acting as antimicrobial agents and by regulating leukocyte recruitment to sites of infection or injury.
- Pathways
S100A9 and S100A8 participate in key inflammation- and immune-related signaling cascades such as the NF-κB pathway and the TLR4 signaling pathway. In these contexts they interact with associated proteins like TLR4 and RAGE (Receptor for Advanced Glycation End-products). Their involvement in these pathways highlights their role in amplifying inflammatory signals during immune responses promoting the expression of pro-inflammatory cytokines.
- Associated diseases and disorders
S100A9 and S100A8 links with conditions like rheumatoid arthritis and inflammatory bowel disease. The upregulation of these proteins often correlates with increased inflammation and severity in such diseases. Moreover they connect with joint damage observed in rheumatoid arthritis through their interaction with RAGE. In inflammatory bowel disease their elevated levels serve as biomarkers for disease activity and guide therapeutic strategies aimed at modulating the immune response.
Product promise
We are dedicated to supporting your work with high quality reagents and we are here for you every step of the way should you need us.
In the unlikely event of one of our products not working as expected, you are covered by our product promise.
Full details and terms and conditions can be found here:
Terms & Conditions.
6 product images
Sandwich ELISA - Human Calprotectin (S100A8/S100A9) ELISA Kit (ab320045)
50 ng/mL of recombinant human S100A8 and S100A9 were tested individually for cross-reactivity. Percent reactivity was determined by interpolating the protein concentration from the human Calprotectin standard curve and expressing the interpolated concentration as a percentage of the protein concentration tested.
Sandwich ELISA - Human Calprotectin (S100A8/S100A9) ELISA Kit (ab320045)
Interpolated concentration of native Calprotectin was measured in duplicate at different sample concentrations. Undiluted samples are as follows: serum 1:4,000, plasma (citrate) 1:4,000, and saliva 1:2,000. The interpolated dilution factor corrected values (to neat sample) are plotted (mean +/- SD, n=2). Sample dilutions are made in 1X Wash Buffer PT.
Sandwich ELISA - Human Calprotectin (S100A8/S100A9) ELISA Kit (ab320045)
Interpolated concentration of native Calprotectin was measured in duplicate at different sample concentrations. Undiluted samples are as follows: plasma (EDTA) 1:1,000, milk 1:500, stimulated PBMC supernatant 1:2,000, and urine 1:500. Human PBMCs were cultured for 46 hours in the presence of 1.5% PHA-M. The interpolated dilution factor corrected values (to neat sample) are plotted (mean +/- SD, n=2). Sample dilutions are made in 1X Wash Buffer PT.
Sandwich ELISA - Human Calprotectin (S100A8/S100A9) ELISA Kit (ab320045)
Standard curve comparison between Human Calprotectin SimpleStep ELISA kit (ab320045) and traditional ELISA kit from leading competitor. The SimpleStep ELISA kit shows increased sensitivity.
Sandwich ELISA - Human Calprotectin (S100A8/S100A9) ELISA Kit (ab320045)
Example of human Calprotectin standard curve. Background-subtracted data values (mean +/- SD) are graphed.
Sandwich ELISA - Human Calprotectin (S100A8/S100A9) ELISA Kit (ab320045)
Interpolated concentration of native Calprotectin was measured in duplicate at different sample concentrations. Undiluted samples are as follows: HL-60 cell extract 500 ng/mL and colon tissue extract 200 ng/mL. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). Sample dilutions are made in 1X Wash Buffer PT.
Downloads
Product protocols
- Visit the general protocols
- Visit troubleshooting
Please note: All products are 'FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC OR THERAPEUTIC PROCEDURES'.
For licensing inquiries, please contact partnerships@abcam.com