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resDetect™ Universal Protein A Quick ELISA Kit (Boiling-free)

For research use only.
Product Details
Assay TypeSandwich-ELISA
AnalyteProtein A
Format96T
Regulatory StatusRUO
Sensitivity<40pg/mL
Standard Curve Range50 pg/mL-1600 pg/mL
Assay Time2 hr
Suitable Sample TypeFor the quantitative determination of recombinant Protein A, alkaline-resistant Protein A and MaXtar® ARPA ligand Protein A.
Sample volume25 μL
Materials Provided
IDComponentsSize
RES029-C01Pre-Coated Anti-Protein A Antibody Microplate1 plate
RES029-C02AAlkali-Tolerant Recombinant Protein A Standard (1μg/mL)100 μL
RES029-C02BMaXtar® ARPA ligand Protein A Standard (Bio-Link Co.) (1μg/mL)100 μL
RES029-C03Recombinant Protein A Standard (1μg/mL)100 μL
RES029-C04Biotin-Anti-Protein A Antibody1.5 mL
RES029-C05Streptavidin-HRP10 μg
RES029-C0610×Sample Dilution Buffer15 mL
RES029-C07Denaturation Buffer15 mL
RES029-C0820×Washing Buffer30 mL
RES029-C09Antibody Dilution Buffer15 mL
RES029-C10Streptavidin-HRP Dilution Buffer15 mL
RES029-C11Substrate Solution12 mL
RES029-C12Stop Solution8 mL
  • Background
    Protein A is a cell wall protein of Staphylococcus aureus, it has a variety of specific biological characteristics. Due to its high affinity with the Fc part of certain immunoglobulins (especially IgG), it is widely used in the purification of biopharmaceuticals (such as antibodies, vaccines, etc.). However, during the purification, protein A may leach from the purification column and result in contamination of the antibody drugs prepared. Once the remaining protein A enters the human body, it will easily activate the immune response of the organism, and there is a safety risk, so there are strict regulations on the residual level of Protein A in antibody drug preparations. Therefore, the detection of residual Protein A in antibody drugs purified from Protein A purification column is a key quality control step in the production process of antibody drug preparations.

    The resDetect™ Universal Protein A Quick ELISA Kit (Boiling-free) can detect protein A or unnatural protein A variants within 2 hours, it is high sensitive and easy to operate because the sample processing has been changed from the original boiling method to adding denaturing reagents. The sample processing process has been shortened by one hour. Whether in upstream small-scale trials or downstream large-scale of antibody production processes, this kit can help you to accurate analysis of samples, monitor the protein A levels and ensure product quality.

  • Application

    The kit is developed for the detection of natural or structurally conserved recombinant forms of Protein A and alkaline-resistant Protein A variants, such as MabSelect SuReTM , MaXtar® ARPA ligand (Bio-Link Co.), UniMab® 50 Protein A (NanoMicro) etc. in bioprocess manufacturing applications. It is used as a universal protein A and variants ligand detection tool to aid in optimal antibody purification process development and in routine quality control of in-process streams as well as final product.

    It is for research use only.

  • Storage
    Unopened kit should be stored at 2°C-8°C upon receiving.

    Find the expiration date on the outside packaging and do not use reagents past their expiration date.

    The opened kit should be stored per components table. The shelf life is 30 days from the date of opening.

  • Assay Principles
    The resDetect™ Universal Protein A Quick ELISA Kit (Boiling-free) is used to measure the levels of protein A and protein A variants by employing a standard sandwich-ELISA format. The micro-plate in the kit has been pre-coated with anti-protein A polyclonal antibody. Firstly, the standard samples provided in kit and your samples are treated with Denaturation Buffer to dissociation of protein A and antibody, stand a few minute. Before adding standards and samples, add the Biotin-Anti-Protein A Antibody to the plate to ensure that the standard samples are neutralized by the Biotin-Anti-Protein A Antibody buffer solution and protect the pre-coated antibody on the plate. Then, add the standard samples and your samples to the plate and form Antibody-antigen (Protein A) - biotinylated antibody complex, incubate and wash the wells. Next add Horseradish peroxidase conjugated streptavidin (Streptavidin-HRP) to the plate, incubate and wash the wells to remove any unbound reactants. At last, load the tetramethylbenzidine (TMB) substrate into the wells and monitor a blue color. The reaction is stopped by the addition of a stop solution and the color turns yellow. The intensity of the absorbance can be measured at 450nm and 630nm on a microtiter plate reader. The OD Value reflects the amount of protein A.
Typical Data Please refer to DS document for the assay protocol.
 protein A (SuRe) TYPICAL DATA

Detection of Recombinant Protein A by sandwich-ELISA Assay.
Immobilized Anti-Protein A Antibody can bind Recombinant Protein A. Detection was performed using Biotin-Anti-Protein A Antibody with sensitivity of 50 pg/mL (QC tested). For each experiment, a standard curve needs to be set for each micro-plate, and the specific OD value may vary depending on different laboratories, testers, or equipments. The following example data is for reference only.

 protein A (SuRe) TYPICAL DATA

Detection of Alkali-Tolerant Recombinant Protein A by sandwich-ELISA Assay.
Immobilized Anti-Protein A Antibody can bind Alkali-Tolerant Recombinant Protein A. Detection was performed using Biotin-Anti-Protein A Antibody with sensitivity of 50 pg/mL (QC tested). For each experiment, a standard curve needs to be set for each micro-plate, and the specific OD value may vary depending on different laboratories, testers, or equipments. The following example data is for reference only.

 protein A (SuRe) TYPICAL DATA

Detection of MaXtar® ARPA ligand Protein A (Bio-Link Co.) by sandwich-ELISA Assay.
Immobilized Anti-Protein A Antibody can bind MaXtar® ARPA ligand Protein A (Bio-Link Co.). Detection was performed using Biotin-Anti-Protein A Antibody with sensitivity of 50 pg/mL (QC tested). For each experiment, a standard curve needs to be set for each micro-plate, and the specific OD value may vary depending on different laboratories, testers, or equipments. The following example data is for reference only.

  • Clinical and Translational Updates

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