Human CD59 ELISA Kit (ab263893)
Key features and details
- One-wash 90 minute protocol
- Sensitivity: 4.9 pg/ml
- Range: 23.44 pg/ml - 1500 pg/ml
- Sample type: Cell culture supernatant, Cit plasma, EDTA Plasma, Hep Plasma, Serum, Tissue Extracts, Urine
- Detection method: Colorimetric
- Assay type: Sandwich (quantitative)
- Reacts with: Human
Overview
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Product name
Human CD59 ELISA Kit
See all CD59 kits -
Detection method
Colorimetric -
Precision
Intra-assay Sample n Mean SD CV% Serum 8 7.5% Inter-assay Sample n Mean SD CV% Serum 3 6.4% -
Sample type
Cell culture supernatant, Urine, Serum, Tissue Extracts, Hep Plasma, EDTA Plasma, Cit plasma -
Assay type
Sandwich (quantitative) -
Sensitivity
4.9 pg/ml -
Range
23.44 pg/ml - 1500 pg/ml -
Recovery
Sample specific recovery Sample type Average % Range Cell culture supernatant 112 107% - 116% Urine 111 107% - 120% Serum 90 82% - 95% Tissue Extracts 99 93% - 109% Hep Plasma 100 84% - 109% EDTA Plasma 92 85% - 97% Cit plasma 94 80% - 117% -
Assay time
1h 30m -
Assay duration
One step assay -
Species reactivity
Reacts with: Human
Does not react with: Cow -
Product overview
Human CD59 ELISA Kit (ab263893) is a single-wash 90 min sandwich ELISA designed for the quantitative measurement of CD59 protein in cell culture supernatant, cit plasma, edta plasma, hep plasma, serum, urine, and tissue extracts. It uses our proprietary SimpleStep ELISA® technology. Quantitate Human CD59 with 4.9 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
CD59 is a potent inhibitor of the complement membrane attack complex (MAC). It acts by binding to C8 and/or C9 of the assembling MAC, thereby preventing incorporation of the multiple copies of C9 required for complete formation of the osmolytic pore. This inhibitor appears to be species-specific. Involved in signal transduction for T-cell activation complexed to a protein tyrosine kinase. The soluble form from urine retains its specific complement binding activity but exhibits greatly reduced ability to inhibit MAC assembly on cell membranes. Glycated CD59 is found in diabetic subjects, but only at minimal levels in nondiabetic subjects. Glycated CD59 lacks MAC-inhibitory function and confers to vascular complications of diabetes. Human CD59 shares 46%, 44%, and 47% sequence homology with mouse, rat, and bovine, respectively.
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. -
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 CD59 Capture Antibody 1 x 600µl 10X Human CD59 Detector Antibody 1 x 600µl 10X Wash Buffer PT (ab206977) 1 x 20ml 50X Cell Extraction Enhancer Solution (ab193971) 1 x 1ml 5X Cell Extraction Buffer PTR (ab193970) 1 x 10ml Antibody Diluent 5BI 1 x 6ml Human CD59 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
Potent inhibitor of the complement membrane attack complex (MAC) action. Acts by binding to the C8 and/or C9 complements of the assembling MAC, thereby preventing incorporation of the multiple copies of C9 required for complete formation of the osmolytic pore. This inhibitor appears to be species-specific. Involved in signal transduction for T-cell activation complexed to a protein tyrosine kinase.
The soluble form from urine retains its specific complement binding activity, but exhibits greatly reduced ability to inhibit MAC assembly on cell membranes. -
Involvement in disease
Defects in CD59 are the cause of CD59 deficiency (CD59D) [MIM:612300]. -
Sequence similarities
Contains 1 UPAR/Ly6 domain. -
Post-translational
modificationsN- and O-glycosylated. The N-glycosylation mainly consists of a family of biantennary complex-type structures with and without lactosamine extensions and outer arm fucose residues. Also significant amounts of triantennary complexes (22%). Variable sialylation also present in the Asn-43 oligosaccharide. The predominant O-glycans are mono-sialylated forms of the disaccharide, Gal-beta-1,3GalNAc, and their sites of attachment are probably on Thr-76 and Thr-77. The GPI-anchor of soluble urinary CD59 has no inositol-associated phospholipid, but is composed of seven different GPI-anchor variants of one or more monosaccharide units. Major variants contain sialic acid, mannose and glucosamine Sialic acid linked to an N-acetylhexosamine-galactose arm is present in two variants.
Glycated. Glycation is found in diabetic subjects, but only at minimal levels in nondiabetic subjects. Glycated CD59 lacks MAC-inhibitory function and confers to vascular complications of diabetes. -
Cellular localization
Cell membrane. Secreted. Soluble form found in a number of tissues. - Information by UniProt
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Alternative names
- 16.3A5
- 1F5
- 1F5 antigen
see all -
Database links
- Entrez Gene: 966 Human
- Omim: 107271 Human
- SwissProt: P13987 Human
- Unigene: 278573 Human
- Unigene: 709466 Human
- Unigene: 710641 Human
Images
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SimpleStep ELISA Protocol Diagram
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 CD59 standard curve in Sample Diluent NS + 1X Enhancer.The CD59 standard curve was prepared as described in Section 10. Raw data values are shown in the table. Background-subtracted data values (mean +/- SD) are graphed.
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Interpolated concentrations of native CD59 in human serum and plasma samples.The concentrations of CD59 were measured in duplicates, interpolated from the CD59 standard curves and corrected for sample dilution. Undiluted samples are as follows: serum 12.5%, plasma (citrate) 12.5%, plasma (EDTA) 12.5%, and plasma (heparin) 12.5%. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean CD59 concentration was determined to be 12.87 ng/mL in serum, 11.50 ng/mL in plasma (citrate), 9.08 ng/mL in plasma (EDTA), and 11.63 ng/mL in plasma (heparin).
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Interpolated concentrations of native CD59 in human urine and cell culture supernatant samples.The concentrations of CD59 were measured in duplicates, interpolated from the CD59 standard curves and corrected for sample dilution. Undiluted samples are as follows: urine 1:3200 and HUVEC supernatant 12.5%. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean CD59 concentration was determined to be 1328.22 ng/mL in urine and 8.97 ng/mL in HUVEC supernatant.
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Interpolated concentrations of native CD59 in HeLa, HUVEC, and liver cell and tissue culture extracts based on a 62.5 µg/mL, 25 µg/mL, and 31.25 µg/mL extract load, respectively.The concentrations of CD59 were measured in duplicate and interpolated from the CD59 standard curve and corrected for sample dilution. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean CD59 concentration was determined to be 2.70 ng/mL in HeLa extract, 2.70 ng/mL in HUVEC extract, and 0.77 ng/mL in liver extract.
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6.25% 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 CD59 concentration was determined to be 11.88 ng/mL with a range of 9.37 – 18.19 ng/mL.
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Sandwich ELISA - Human CD59 ELISA Kit (ab263893)To learn more about the advantages of recombinant antibodies see here.
