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  1. Link

    human-alpha-1-microglobulin-elisa-kit-ab226899.pdf

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Cardiovascular Blood Serum Proteins
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RecombinantSimpleStep

Human Alpha-1-Microglobulin ELISA Kit (ab226899)

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  • Protocol Booklet
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Other - Human Alpha-1-Microglobulin ELISA Kit (ab226899)
  • Example of HumanAlpha 1-Microglobulin standard curve in Sample Diluent NS
  • Interpolated concentrations of native Alpha 1-Microglobulin in Human serum and plasma samples
  • Serum from ten individual healthy Human female donors was measured in duplicate
  • Interpolated concentrations of native Alpha 1-Microglobulin in Human cell culture supernatant and urine samples
  • Sandwich ELISA - Human Alpha-1-Microglobulin ELISA Kit (ab226899)

Key features and details

  • One-wash 90 minute protocol
  • Sensitivity: 725 pg/ml
  • Range: 1.56 ng/ml - 100 ng/ml
  • Sample type: Cit plasma, EDTA Plasma, Hep Plasma, Serum, Urine
  • Detection method: Colorimetric
  • Assay type: Sandwich (quantitative)
  • Reacts with: Human

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Protein
Native Human Alpha 1 microglobulin protein (ab96149)

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Overview

  • 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

    Human Alpha-1-Microglobulin ELISA Kit (ab226899) is a single-wash 90 min sandwich ELISA designed for the quantitative measurement of Alpha-1-Microglobulin protein in cit plasma, edta plasma, hep plasma, serum, and urine. It uses our proprietary SimpleStep ELISA® technology. Quantitate Human Alpha-1-Microglobulin with 725 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


    A 384-well SimpleStep ELISA® microplate (ab203359) is available to use as an alternative to the 96-well microplate provided with SimpleStep ELISA® kits.

  • Notes

    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.  

  • Platform

    Pre-coated microplate (12 x 8 well strips)

Properties

  • 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

    • Cardiovascular
    • Blood
    • Serum Proteins
    • Cardiovascular
    • Blood
    • Other
    • Cell Biology
    • Proteolysis / Ubiquitin
    • Protease inhibitors
    • Cancer
    • Tumor biomarkers
    • Other
  • 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
    modifications

    The 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.
  • Target information above from: UniProt accession P02760 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
  • Alternative names

    • A1M
    • Alpha 1 microglobulin/bikunin precursor
    • Alpha 1 microglycoprotein
    • Alpha-1 microglycoprotein
    • Alpha-1-microglobulin
    • AMBP
    • AMBP_HUMAN
    • Bikunin
    • Complex-forming glycoprotein heterogeneous in charge
    • EDC1
    • Growth inhibiting protein 19
    • HCP
    • HI-30
    • HI30
    • IATIL
    • ITI
    • ITI-LC
    • ITIL
    • Microglobulin (alpha 1)
    • Protein AMBP
    • Protein HC
    • Trypstatin
    • Uristatin
    • Uronic-acid-rich protein
    • UTI
    see all
  • Database links

    • Entrez Gene: 259 Human
    • Omim: 176870 Human
    • SwissProt: P02760 Human
    • Unigene: 436911 Human

    Associated products

    • Alternative Versions

      • Human Alpha-1-Microglobulin Antibody Pair - BSA and Azide free (ab244036)

    Images

    • Other - Human Alpha-1-Microglobulin ELISA Kit (ab226899)
      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.

       

    • Example of HumanAlpha 1-Microglobulin standard curve in Sample Diluent NS
      Example of HumanAlpha 1-Microglobulin standard curve in Sample Diluent NS

      Background-subtracted data values (mean +/- SD) are graphed.

    • Interpolated concentrations of native Alpha 1-Microglobulin in Human serum and plasma samples
      Interpolated concentrations of native Alpha 1-Microglobulin in Human serum and plasma samples

      The 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).

    • Serum from ten individual healthy Human female donors was measured in duplicate
      Serum from ten individual healthy Human female donors was measured in duplicate

      Interpolated 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.

    • Interpolated concentrations of native Alpha 1-Microglobulin in Human cell culture supernatant and urine samples
      Interpolated concentrations of native Alpha 1-Microglobulin in Human cell culture supernatant and urine samples

      The 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.

    • Sandwich ELISA - Human Alpha-1-Microglobulin ELISA Kit (ab226899)
      Sandwich ELISA - Human Alpha-1-Microglobulin ELISA Kit (ab226899)
      To learn more about the advantages of recombinant antibodies see here.

    Protocols

    • Protocol Booklet

    Click here to view the general protocols

    Datasheets and documents

    • Datasheet
    • SDS
  • References (0)

    Publishing research using ab226899? Please let us know so that we can cite the reference in this datasheet.

    ab226899 has not yet been referenced specifically in any publications.

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