Human Alpha-1-Microglobulin ELISA Kit (ab226899)
- Datasheet
- References
- Protocols
Overview
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Product name
Human Alpha-1-Microglobulin ELISA Kit
See all Alpha 1 microglobulin kits -
Detection method
Colorimetric -
Precision
Intra-assay Sample n Mean SD CV% Serum 5 3.2% Inter-assay Sample n Mean SD CV% Serum 3 5.8% -
Sample type
Urine, Serum, Hep Plasma, EDTA Plasma, Cit plasma -
Assay type
Sandwich (quantitative) -
Sensitivity
725 pg/ml -
Range
1.56 ng/ml - 100 ng/ml -
Recovery
Sample specific recovery Sample type Average % Range Urine 104 95% - 109% Serum 96 92% - 102% Cell culture media 106 103% - 109% Hep Plasma 92 88% - 96% EDTA Plasma 97 94% - 102% Cit plasma 96 95% - 97% -
Assay time
1h 30m -
Assay duration
One step assay -
Species reactivity
Reacts with: Human
Does not react with: Cow -
Product overview
Alpha 1-Microglobulin in vitro SimpleStep ELISA® (Enzyme-Linked Immunosorbent Assay) kit is designed for the quantitative measurement of Alpha 1-Microglobulin protein in Human serum, plasma, urine, cell and tissue culture supernatant.
The SimpleStep ELISA® employs an affinity tag labeled capture antibody and a reporter conjugated detector antibody which immunocapture the sample analyte in solution. This entire complex (capture antibody/analyte/detector antibody) is in turn immobilized via immunoaffinity of an anti-tag antibody coating the well. To perform the assay, samples or standards are added to the wells, followed by the antibody mix. After incubation, the wells are washed to remove unbound material. TMB substrate is added and during incubation is catalyzed by HRP, generating blue coloration. This reaction is then stopped by addition of Stop Solution completing any color change from blue to yellow. Signal is generated proportionally to the amount of bound analyte and the intensity is measured at 450 nm. Optionally, instead of the endpoint reading, development of TMB can be recorded kinetically at 600 nm.
Alpha 1-microglobulin is expressed in the liver and secreted in the plasma acting as a radical scavenger and degrades heme. Alpha 1-Microglobulin can exist as either a monomer or in complexes with IgA or Albumin. The precursor to Alpha 1-microglobulin (Protein AMBP) is cleaved into three separate functioning proteins (Alpha 1-microglobulin, Bikunin, and Trypstatin). Ab226899 is designed to recognize Alpha-1-microglobin, specificity over the pre-cursor has not been tested.
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Tested applications
Suitable for: Sandwich ELISAmore details -
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 Wash Buffer PT (ab206977) 1 x 20ml Antibody Diluent CPR 1 x 6ml 10X Human Alpha 1-Microglobulin Capture Antibody 1 x 600µl 10X Human Alpha 1-Microglobulin Detector Antibody 1 x 600µl Human Alpha 1-Microglobulin 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
Inter-alpha-trypsin inhibitor inhibits trypsin, plasmin, and lysosomal granulocytic elastase. Inhibits calcium oxalate crystallization.
Trypstatin is a trypsin inhibitor. -
Tissue specificity
Expressed by the liver and secreted in plasma. Alpha-1-microglobulin occurs in many physiological fluids including plasma, urine, and cerebrospinal fluid. Inter-alpha-trypsin inhibitor is present in plasma and urine. -
Sequence similarities
In the N-terminal section; belongs to the calycin superfamily. Lipocalin family.
Contains 2 BPTI/Kunitz inhibitor domains. -
Post-translational
modificationsThe precursor is proteolytically processed into separately functioning proteins.
3-hydroxykynurenine, an oxidized tryptophan metabolite that is common in biological fluids, reacts with Cys-53, Lys-111, Lys-137, and Lys-149 to form heterogeneous polycyclic chromophores including hydroxanthommatin. The reaction by alpha-1-microglobulin is autocatalytic; the human protein forms chromophore even when expressed in insect and bacterial cells. The chromophore can react with accessible cysteines forming non-reducible thioether cross-links with other molecules of alpha-1-microglobulin or with other proteins such as Ig alpha-1 chain C region 'Cys-352'.
Heavy chains are interlinked with bikunin via a chondroitin 4-sulfate bridge to the their C-terminal aspartate.
Addition of glycosaminoglycan chondroitin sulfate, allows cross-linking between the different components. -
Cellular localization
Secreted. - Information by UniProt
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Alternative names
- A1M
- Alpha 1 microglobulin/bikunin precursor
- Alpha 1 microglycoprotein
see all -
Database links
- Entrez Gene: 259 Human
- Omim: 176870 Human
- SwissProt: P02760 Human
- Unigene: 436911 Human
Associated products
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Alternative Versions
Applications
Our Abpromise guarantee covers the use of ab226899 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
| Application | Abreviews | Notes |
|---|---|---|
| Sandwich ELISA | Use at an assay dependent concentration. |
Images
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Other - Human Alpha-1-Microglobulin ELISA Kit (ab226899)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 HumanAlpha 1-Microglobulin standard curve in Sample Diluent NSBackground-subtracted data values (mean +/- SD) are graphed.
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Interpolated concentrations of native Alpha 1-Microglobulin in Human serum and plasma samplesThe concentrations of Alpha 1-Microglobulin were measured in duplicates, interpolated from the Alpha 1-Microglobulin standard curves and corrected for sample dilution. Undiluted samples are as follows: serum 1:200, plasma (EDTA) 1:100, plasma (heparin) 1:100, and plasma (citrate) 1:100. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean Alpha 1-Microglobulin concentration was determined to be 29.7 µg/mL in serum, 20.7 µg/mL in plasma (EDTA), 23.2 µg/mL in plasma (heparin) and 21.2 µg/mL in plasma (citrate).
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Serum from ten individual healthy Human female donors was measured in duplicateInterpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean Alpha 1-Microglobulin concentration was determined to be 27.4 µg/mL with a range of 12.6 – 41.9 µg/mL.
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Interpolated concentrations of native Alpha 1-Microglobulin in Human cell culture supernatant and urine samplesThe concentrations of Alpha 1-Microglobulin were measured in duplicates, interpolated from the Alpha 1-Microglobulin standard curves and corrected for sample dilution. Undiluted samples are as follows: HEPG2 Cell culture supernatant 10% and urine 1:100. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean Alpha 1-Microglobulin concentration was determined to be 1026 ng/mL in HEPG2 cell culture supernatant and 9590 ng/mL in urine.
Protocols
References
ab226899 has not yet been referenced specifically in any publications.
