Overview

  • Product name

    Human Hemoglobin subunit alpha ELISA Kit
    See all Hemoglobin subunit alpha kits
  • Detection method

    Colorimetric
  • Precision

    Intra-assay
    Sample n Mean SD CV%
    Overall 5 3.6%
    Inter-assay
    Sample n Mean SD CV%
    Overall 3 14.2%
  • Sample type

    Cell culture supernatant, Serum, Heparin Plasma, EDTA Plasma, Citrate Plasma, Cerebral Spinal Fluid
  • Assay type

    Sandwich (quantitative)
  • Sensitivity

    131 pg/ml
  • Range

    588 pg/ml - 20000 pg/ml
  • Recovery

    Sample specific recovery
    Sample type Average % Range
    Serum 96 84% - 100%
    Whole Blood 101 81% - 115%
    Heparin Plasma 93 82% - 100%
    EDTA Plasma 94 91% - 98%
    Cerebral Spinal Fluid 104 99% - 109%
    serum free media 88 91% - 94%

  • Assay time

    1h 30m
  • Assay duration

    One step assay
  • Species reactivity

    Reacts with: Human
    Does not react with: Mouse, Rat
  • Product overview

    Hemoglobin subunit alpha in vitro SimpleStep ELISA® (Enzyme-Linked Immunosorbent Assay) kit is designed for the quantitative measurement of Hemoglobin subunit alpha protein in human serum, plasma, cerebral spinal fluid, hemolyzed whole blood lysate and cell 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.

  • Notes

    Hemoglobin (Haemoglobin) subunits combine to form the quaternary structure of total hemoglobin, the oxygen carrying protein in red blood cells. In normal adults hemoglobin consists of 2 alpha and 2 beta subunits. Hemoglobin alpha subunit can be modified by the glucose on the terminal valine of beta subunits resulting in the Hemoglobin A1C molecule. The resulting A1C molecule is used to measure the average glucose levels in blood over time. ab219049 measures the total Hemoglobin and can recognize both the alpha subunit and A1C modified protein.

  • Tested applications

    Suitable for: Sandwich ELISAmore details
  • 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 Human Hemoglobin subunit alpha Capture Antibody 1 x 600µl
    10X Human Hemoglobin subunit alpha Detector Antibody 1 x 600µl
    Human Hemoglobin Lyophilized Purified Protein 2 vials
    Antibody Diluent CPI 1 x 6ml
    10X Wash Buffer PT (ab206977) 1 x 20ml
    TMB Development Solution 1 x 12ml
    Stop Solution 1 x 12ml
    Sample Diluent NS 1 x 50ml
    SimpleStep Pre-Coated 96-Well Microplate (ab206978) 1 unit
    Plate Seals 1 unit
  • Research areas

  • Function

    Involved in oxygen transport from the lung to the various peripheral tissues.
  • Tissue specificity

    Red blood cells.
  • Involvement in disease

    Heinz body anemias
    Alpha-thalassemia
    Alpha(0)-thalassemia is associated with non-immune hydrops fetalis, a generalized edema of the fetus with fluid accumulation in the body cavities due to non-immune causes. Non-immune hydrops fetalis is not a diagnosis in itself but a symptom, a feature of many genetic disorders, and the end-stage of a wide variety of disorders.
    Hemoglobin H disease
  • Sequence similarities

    Belongs to the globin family.
  • Post-translational
    modifications

    The initiator Met is not cleaved in variant Thionville and is acetylated.
  • Information by UniProt
  • Alternative names

    • Alpha 1 globin
    • Alpha globin
    • Alpha one globin
    • Alpha-globin
    • HBA_HUMAN
    • HBA1
    • HBA2
    • Hemoglobin alpha 1
    • Hemoglobin alpha 1 chain
    • Hemoglobin alpha 1 globin chain
    • Hemoglobin alpha 2
    • Hemoglobin alpha chain
    • Hemoglobin subunit alpha
    • MGC126895
    • MGC126897
    see all
  • Database links

Applications

Our Abpromise guarantee covers the use of ab219049 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

  • 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.

     

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

  • The concentrations of Hemoglobin subunit alpha were measured in duplicates, interpolated from the Hemoglobin standard curves and corrected for sample dilution. Undiluted samples are as follows: serum 25%, plasma (heparin) 25%, plasma (EDTA) 12.5%, and whole blood 1:80,000 . The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean Hemoglobin subunit alpha concentration was determined to be 0.07 µg/mL in serum, 0.06 µg/mL in plasma (heparin), 0.11 µg/mL in plasma (EDTA), and 400µg/mL in whole blood.

  • Interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean Hemoglobin subunit alpha concentration was determined to be 34 ng/mL with a range of 8 – 137 ng/mL.

  • The concentrations of Hemoglobin subunit alpha were measured in duplicates, interpolated from the Hemoglobin standard curves and corrected for sample dilution. Undiluted samples are as follows: Cerebral spinal fluid 6.25%. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean Hemoglobin subunit alpha concentration was determined to be 540 ng/mL in cerebral spinal fluid.

  • The concentrations of Hemoglobin subunit alpha were measured in duplicates, interpolated from the Hemoglobin standard curves and corrected for sample dilution. Undiluted samples are as follows: Serum free cell culture media 25%. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2).

  • Each protein was loaded at 5ng/mL and interpolated off the hemoglobin standard curve. The interpolated values are plotted (mean +/- SD, n=2). The observed 4-fold difference is expected, due to the size difference between the alpha subunit and the whole molecule.

Protocols

References

ab219049 has not yet been referenced specifically in any publications.

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