Human Haptoglobin ELISA Kit (ab219048)
Key features and details
- One-wash 90 minute protocol
- Sensitivity: 86 pg/ml
- Range: 312.5 pg/ml - 20000 pg/ml
- Sample type: Cell culture supernatant, Cit plasma, EDTA Plasma, Hep Plasma, Saliva, Serum, Urine
- Detection method: Colorimetric
- Assay type: Sandwich (quantitative)
- Reacts with: Human
Overview
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Product name
Human Haptoglobin ELISA Kit
See all Haptoglobin kits -
Detection method
Colorimetric -
Precision
Intra-assay Sample n Mean SD CV% Serum 5 1.8% Inter-assay Sample n Mean SD CV% Serum 3 4.4% -
Sample type
Cell culture supernatant, Saliva, Urine, Serum, Hep Plasma, EDTA Plasma, Cit plasma -
Assay type
Sandwich (quantitative) -
Sensitivity
86 pg/ml -
Range
312.5 pg/ml - 20000 pg/ml -
Recovery
Sample specific recovery Sample type Average % Range Cell culture supernatant 104 97% - 113% Saliva 105 101% - 113% Urine 102 101% - 105% Serum 107 104% - 113% Hep Plasma 104 99% - 110% EDTA Plasma 105 99% - 111% Cit plasma 107 102% - 120% -
Assay time
1h 30m -
Assay duration
One step assay -
Species reactivity
Reacts with: Human
Does not react with: Cow -
Product overview
Human Haptoglobin ELISA Kit (ab219048) is a single-wash 90 min sandwich ELISA designed for the quantitative measurement of Haptoglobin protein in cell culture supernatant, cit plasma, edta plasma, hep plasma, saliva, serum, and urine. It uses our proprietary SimpleStep ELISA® technology. Quantitate Human Haptoglobin with 86 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 stripsA 384-well SimpleStep ELISA® microplate (ab203359) is available to use as an alternative to the 96-well microplate provided with SimpleStep ELISA® kits.
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Notes
Haptoglobin binds free hemoglobin to allow recycling of heme iron and prevent kidney damage. The hemoglobin/haptoglobin complex is then cleared by the Kupfer cells through endocytosis, via the CD163 scavenger receptor. The precursor to haptoglobin, zonulin, is cleaved into alpha and beta chains that homo and heterodimerize to form haptoglobin. In the population, there are two major allelic forms of hemoglobin that give rise to 3 major phenotypes (1-1, 2-1, and 2-2), which are a combination of the homo and heterodimerized alpha and beta chains. This kit recognizes both Isoform 1 and 2 of the alpha chain.
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Platform
Pre-coated microplate (12 x 8 well strips)
Properties
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Storage instructions
Store at +4°C. Please refer to protocols. -
Components 1 x 96 tests 10X Human Haptoglobin Capture Antibody 1 x 600µl 10X Human Haptoglobin Detector Antibody 1 x 600µl 10X Wash Buffer PT (ab206977) 1 x 20ml Antibody Diluent 4BI 1 x 6ml Human Haptoglobin Lyophilized Recombinant Protein 2 vials Plate Seals 1 unit Sample Diluent NS (ab193972) 1 x 50ml SimpleStep Pre-Coated 96-Well Microplate (ab206978) 1 unit Stop Solution 1 x 12ml TMB Development Solution 1 x 12ml -
Research areas
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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 Antimicrobial; Antioxidant, has antibacterial activity and plays a role in modulating many aspects of the acute phase response. Hemoglobin/haptoglobin complexes are rapidely cleared by the macrophage CD163 scavenger receptor expressed on the surface of liver Kupfer cells through an endocytic lysosomal degradation pathway.
Uncleaved haptoglogin, also 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. -
Tissue specificity
Expressed by the liver and secreted in plasma. -
Involvement in disease
Anhaptoglobinemia -
Sequence similarities
Belongs to the peptidase S1 family.
Contains 1 peptidase S1 domain.
Contains 2 Sushi (CCP/SCR) domains. -
Cellular localization
Secreted. - Information by UniProt
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Alternative names
- Binding peptide
- BP
- Haptoglobin alpha chain
see all -
Database links
- Entrez Gene: 3240 Human
- Omim: 140100 Human
- SwissProt: P00738 Human
- Unigene: 513711 Human
- Unigene: 702099 Human
Images
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Other - Human Haptoglobin ELISA Kit (ab219048)
SimpleStep ELISA technology allows the formation of the antibody-antigen complex in one single step, reducing assay time to 90 minutes. Add samples or standards and antibody mix to wells all at once, incubate, wash, and add your final substrate. See protocol for a detailed step-by-step guide.
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Example of human haptoglobin standard curve.Background-subtracted data values (mean +/- SD) are graphed.
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Human Haptoglobin standard curve comparison dataStandard curve comparison between human haptoglobin SimpleStep ELISA® kit and traditional ELISA kit from leading competitor. SimpleStep ELISA kit shows comparable sensitivity.
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Interpolated concentrations of native haptoglobin in human serum, and plasma samples.The concentrations of haptoglobin were measured in duplicates, interpolated from the haptoglobin standard curves and corrected for sample dilution. Undiluted samples are as follows: serum 1:2000, plasma (citrate) 1:1000, plasma (EDTA) 1:1000, and plasma (heparin) 1:2000. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean haptoglobin concentration was determined to be 8534 ng/mL in serum, 3624 ng/mL in plasma (citrate), 4154 ng/mL in plasma (EDTA) and 8087 ng/mL in plasma (heparin).
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Serum from 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 6.5 µg/mL with a range of 0.15 – 25.3 µg/mL.
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Interpolated concentrations of native haptoglobin in human urine, saliva, and HEPG2 cell culture supernatant samples.The concentrations of haptoglobin were measured in duplicates, interpolated from the haptoglobin standard curves and corrected for sample dilution. 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). The mean haptoglobin concentration was determined to be 12.6 ng/mL in urine, 27.8 ng/mL in saliva, and 66.9ng/mL in HEPG2 cell culture supernatant.
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Sandwich ELISA - Human Haptoglobin ELISA Kit (ab219048)To learn more about the advantages of recombinant antibodies see here.
