Human Haptoglobin ELISA Kit is a single-wash 90-min Simplestep used to quantify Human Haptoglobin with a sensitivity of 51.9 ng/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
- Design your own immunoassay: we also offer the conjugation-ready antibody pair
Images
Key facts
- Detection method
- Colorimetric
- Sample types
- Serum, Heparin Plasma, Citrate plasma, EDTA Plasma, Urine, Saliva, Cell culture supernatant
- Assay type
- Sandwich (quantitative)
- Reactive species
- Human
- Range
- 156 - 10000 ng/mL
- Assay time
- 1h 30m
- Sensitivity
- = 51.9 ng/mL
Reactivity data
| Application | Reactivity | Dilution info | Notes |
|---|---|---|---|
Application sELISA | Reactivity Reacts | Dilution info - | Notes - |
Target data
Function
As a result of hemolysis, hemoglobin is found to accumulate in the kidney and is secreted in the urine. Haptoglobin captures, and combines with free plasma hemoglobin to allow hepatic recycling of heme iron and to prevent kidney damage. Haptoglobin also acts as an antioxidant, has antibacterial activity, and plays a role in modulating many aspects of the acute phase response. Hemoglobin/haptoglobin complexes are rapidly cleared by the macrophage CD163 scavenger receptor expressed on the surface of liver Kupfer cells through an endocytic lysosomal degradation pathway. The uncleaved form of allele alpha-2 (2-2), known as zonulin, plays a role in intestinal permeability, allowing intercellular tight junction disassembly, and controlling the equilibrium between tolerance and immunity to non-self antigens.
Alternative names
Haptoglobin, Zonulin, HP
What's included?
Recommended products
Human Haptoglobin ELISA Kit is a single-wash 90-min Simplestep used to quantify Human Haptoglobin with a sensitivity of 51.9 ng/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
- Design your own immunoassay: we also offer the conjugation-ready antibody pair
Key facts
- Detection method
- Colorimetric
- Sample types
- Serum, Heparin Plasma, Citrate plasma, EDTA Plasma, Urine, Saliva, Cell culture supernatant
- Assay type
- Sandwich (quantitative)
- Reactive species
- Human
- Range
- 156 - 10000 ng/mL
- Assay time
- 1h 30m
- Assay Platform
- Pre-coated microplate (12 x 8 well strips)
- Sensitivity
- = 51.9 ng/mL
- General recovery
- 107%
Precision
Intra assay
| Sample | n | C.V. |
|---|---|---|
Sample Serum | n 5 | C.V. 1.8 |
Inter assay
| Sample | n | C.V. |
|---|---|---|
Sample Serum | n 3 | C.V. 4.4 |
Recovery
Sample specific recovery
| Sample type | Average % | Range |
|---|---|---|
Sample type Serum | Average % = 107 | Range 104 - 113 % |
Sample type Heparin Plasma | Average % = 105 | Range 99 - 111 % |
Sample type Citrate plasma | Average % = 104 | Range 99 - 110 % |
Sample type EDTA Plasma | Average % = 107 | Range 102 - 120 % |
Sample type Urine | Average % = 102 | Range 101 - 105 % |
Sample type Saliva | Average % = 105 | Range 101 - 113 % |
Sample type Cell culture supernatant | Average % = 104 | Range 97 - 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 Haptoglobin ELISA Kit ab309180 is a rapid single-wash 90-min Sandwich ELISA to measure Human Haptoglobin in cell culture supernatant, citrate plasma, EDTA plasma, heparin plasma, saliva, serum, urine. This SimpleStep sensitivity is 51.9 ng/mL.
How the assay works
Human Haptoglobin SimpleStep ELISA®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.
Assay Specificity
Our SimpleStep ELISA® kits use recombinant monoclonal antibodies rigorously validated to ensure the highest level of consistency and reproducibility, improved sensitivity and specificity and ease of scalability and security of supply.
Please refer to our protocol booklet for more details.
Human Haptoglobin ELISA Kit ab309180 protocol summary
1. Mix: add samples/standards to the wells together with the capture and detector antibody cocktail. Incubate 1 hr at room temperature
2. Wash
3. Add TMB development solution - incubate for 10 min
4. Add Stop solution
5. Read the results on a plate reader at 450 nm
Supplementary info
- This supplementary information is collated from multiple sources and compiled automatically.
- Activity summary
Haptoglobin (Hp) also known as Hp protein is a glycoprotein with a molecular weight ranging from 85 to 100 kDa varying due to its ability to form different dimeric and multimeric structures. It is primarily synthesized in the liver and circulating in the plasma. Mechanically haptoglobin binds free hemoglobin released from erythrocytes preventing oxidative damage and hemoglobin-driven deleterious effects. Haptoglobin's binding to hemoglobin forms a haptoglobin-hemoglobin complex that gets cleared by the CD163 receptor on macrophages mainly in the liver and spleen.
- Biological function summary
Haptoglobin functions as an antioxidant and an acute phase protein playing roles in inflammation and immune response. It is an essential component of the antioxidant defense system reducing oxidative stress by binding free hemoglobin. Additionally haptoglobin can participate in complex formation with hemoglobin to facilitate the clearance of excess free hemoglobin from circulation. Its expression increases during inflammation under the regulation of cytokines like IL-6.
- Pathways
The haptoglobin protein is part of the hemoglobin clearance pathway integral for iron metabolism and homeostasis. This pathway ensures the safe removal of hemoglobin from the blood helping to prevent kidney damage. Haptoglobin is related to other proteins such as CD163 and the liver-derived hepatocellular proteins involved in the clearance process. Also haptoglobin is linked to antioxidant pathways collaborating with oxidative stress regulators to manage reactive oxygen species levels.
- Associated diseases and disorders
Changes in haptoglobin levels are associated with hemolytic anemia and inflammatory conditions. Low haptoglobin levels often suggest hemolytic anemia due to its rapid binding with free hemoglobin in the bloodstream. During inflammation and infections 'anti-Hp' tests (haptoglobin tests) may show increased haptoglobin levels as a response to cytokine signaling. Moreover haptoglobin polymorphisms have a connection with ischemic stroke susceptibility where its isoforms show differing efficiencies in binding free hemoglobin. These findings highlight haptoglobin's relevance as a biomarker for these conditions and illustrate its interaction with hematologic and immune proteins in disease contexts.
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.
5 product images
ELISA - Human Haptoglobin ELISA Kit (ab309180)
Sandwich ELISA - Human Haptoglobin ELISA Kit (ab309180)
Serum of ten individual healthy human male donors was measured in duplicate. Interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean Haptoglobin concentration was determined to be 3.25 mg/mL with a range of 0.075 – 12.65 mg/mL.
Sandwich ELISA - Human Haptoglobin ELISA Kit (ab309180)
Example of human Haptoglobin standard curve. Background-subtracted data values (mean +/- SD) are graphed.
Sandwich ELISA - Human Haptoglobin ELISA Kit (ab309180)
Interpolated concentration of native Haptoglobin was measured in duplicate at different sample concentrations. Undiluted samples are as follows: urine 25%, saliva 50%, and HepG2 cell culture supernatant 12.5%. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). Sample dilutions are made in Sample Diluent NS.
Sandwich ELISA - Human Haptoglobin ELISA Kit (ab309180)
Interpolated concentration of native Haptoglobin was measured in duplicate at different sample concentrations. Undiluted samples are as follows: serum 1:2,000, plasma – Citrate 1:1,000, plasma – EDTA 1:1,000, and plasma – Heparin 1:2,000. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). Sample dilutions are made in Sample Diluent NS.
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