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IgG1 ELISA Kit

Reaktivität: Human Colorimetric Sandwich ELISA 0.05-40 ng/mL Cell Culture Supernatant, Plasma, Serum
Produktnummer ABIN625006
  • Target Alle IgG1 ELISA Kits anzeigen
    IgG1
    Reaktivität
    • 7
    • 3
    • 2
    • 2
    • 2
    • 2
    • 1
    • 1
    • 1
    • 1
    • 1
    • 1
    Human
    Nachweismethode
    Colorimetric
    Methodentyp
    Sandwich ELISA
    Detektionsbereich
    0.05-40 ng/mL
    Untere Nachweisgrenze
    0.05 ng/mL
    Applikation
    ELISA
    Verwendungszweck
    Human IgG1 ELISA Kit for cell culture supernatants, plasma, and serum samples.
    Proben
    Plasma, Cell Culture Supernatant, Serum
    Analytische Methode
    Quantitative
    Spezifität
    Detect human IgG1. No cross reactivity was found againt IgG2, IgG3, IgG4, IgM, IgA, IgD and IgE
    Sensitivität
    50 pg/mL
    Produktmerkmale
    • Strip plates and additional reagents allow for use in multiple experiments
    • Quantitative protein detection
    • Establishes normal range
    • The best products for confirmation of antibody array data
    Bestandteile
    • Pre-Coated 96-well Strip Microplate
    • Wash Buffer
    • Stop Solution
    • Assay Diluent(s)
    • Lyophilized Standard
    • Biotinylated Detection Antibody
    • Streptavidin-Conjugated HRP
    • TMB One-Step Substrate
    Benötigtes Material
    • Distilled or deionized water
    • Precision pipettes to deliver 2 μL to 1 μL volumes
    • Adjustable 1-25 μL pipettes for reagent preparation
    • 100 μL and 1 liter graduated cylinders
    • Tubes to prepare standard and sample dilutions
    • Absorbent paper
    • Microplate reader capable of measuring absorbance at 450nm
    • Log-log graph paper or computer and software for ELISA data analysis
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  • Applikationshinweise
    Recommended Dilution for serum and plasma samples~10,000,000 fold
    Probenmenge
    100 μL
    Plattentyp
    Pre-coated
    Protokoll
    1. Prepare all reagents, samples and standards as instructed in the manual.
    2. Add 100 μL of standard or sample to each well.
    3. Incubate 2.5 h at RT or O/N at 4 °C.
    4. Add 100 μL of prepared biotin antibody to each well.
    5. Incubate 1 h at RT.
    6. Add 100 μL of prepared Streptavidin solution to each well.
    7. Incubate 45 min at RT.
    8. Add 100 μL of TMB One-Step Substrate Reagent to each well.
    9. Incubate 30 min at RT.
    10. Add 50 μL of Stop Solution to each well.
    11. Read at 450 nm immediately.
    Aufbereitung der Reagenzien
    1. Bring all reagents and samples to room temperature (18 - 25 °C) before use.
      2. Sample dilution: 1x Assay Diluent (Item E) should be used for dilution of serum/plasma samples. Suggested dilution for normal serum/plasma: 10,000,000 fold. For example, add 1 µL of serum/plasma into a tube with 99 µL 1x Assay Diluent to prepare a 100-fold diluted sample. Mix through and then pipette 1 µL of prepared 100-fold diluted sample into a tube with 99 µL 1x Assay Diluent to prepare a 10,000 fold diluted sample. Mix through and then pipette 1 µL of prepared 10,000-fold diluted sample into a tube with 999 µL 1x Assay Diluent to prepare a final 10,000,000 fold diluted sample. Please note that levels of the target protein may vary between different specimens. Optimal dilution factors for each sample must be determined by the investigator.
      3. Assay Diluent should be diluted 5-fold with deionized or distilled water before use.
      4. Preparation of standard: Briefly spin the vial of Item C and then add 600 µL 1x Assay Diluent into Item C vial to prepare a 300 ng/mL stock standard solution. Dissolve the powder thoroughly by a gentle mix. Add 80 µL IgG1 standard (300 ng/mL) from the vial of Item C, into a tube with 520 µL 1x Assay Diluent to prepare a 40 ng/mL standard. Pipette 400 µL 1x Assay Diluent into each tube. Use the stock standard solution to produce a dilution series . Mix each tube thoroughly before the next transfer. 1x Assay Diluent serves as the zero standard (0 ng/mL). 80 µL standard + 520 µL 200 µL 200 µL 200 µL 200 µL 200 µL 200myl 40 13.3 4.44 1.48 0.49 0.16 0.05 0 ng/mL ng/mL ng/mL ng/mL ng/mL ng/mL ng/mL ng/mL
      5. If the Wash Concentrate (20x) (Item B) contains visible crystals, warm to room temperature and mix gently until dissolved. Dilute 20 ml of Wash Buffer Concentrate into deionized or distilled water to yield 400 ml of 1x Wash Buffer.
      6. Briefly spin the Detection Antibody vial (Item F) before use. Add 100 µL of 1x Assay Diluent into the vial to prepare a detection antibody concentrate. Pipette up and down to mix gently (the concentrate can be stored at 4 °C for 5 days). The detection antibody concentrate should be diluted 80-fold with 1x Assay Diluent and used in step 4 of Part VI Assay Procedure.
      7. Briefly spin the HRP-Streptavidin concentrate vial (Item G) ) and pipette up and down to mix gently before use. HRP-Streptavidin concentrate should be diluted 2,000-fold with 1x Assay Diluent. For example: Briefly spin the vial (Item G) and pipette up and down to mix gently . Add 6 µL of HRP-Streptavidin concentrate into a tube with 12 ml 1x Assay Diluent to prepare a final 2,000 fold diluted HRP- Streptavidin solution (don't store the diluted solution for next day use). Mix well.
    Testdurchführung
    1. Bring all reagents and samples to room temperature (18 - 25 °C) before use. It is recommended that all standards and samples be run at least in duplicate.
      2. Add 100 µL of each standard (see Reagent Preparation step 2) and sample into appropriate wells. Cover well and incubate for 2.5 hours at room temperature or over night at 4 °C with gentle shaking.
      3. Discard the solution and wash 4 times with 1x Wash Solution. Wash by filling each well with Wash Buffer (300 myl) using a multi-channel Pipette or autowasher. Complete removal of liquid at each step is essential to good performance. After the last wash, remove any remaining Wash Buffer by aspirating or decanting. Invert the plate and blot it against clean paper towels.
      4. Add 100 µL of 1x prepared biotinylated antibody (Reagent Preparation step 6) to each well. Incubate for 1 hour at room temperature with gentle shaking.
      5. Discard the solution. Repeat the wash as in step
      6. Add 100 µL of prepared Streptavidin solution (see Reagent Preparation step 7) to each well. Incubate for 45 minutes at room temperature with gentle shaking.
      7. Discard the solution. Repeat the wash as in step
      8. Add 100 µL of TMB One-Step Substrate Reagent (Item H) to each well. Incubate for 30 minutes at room temperature in the dark with gentle shaking.
      9. Add 50 µL of Stop Solution (Item I) to each well. Read at 450 nm immediately.
    Ergebnisberechnung

    Calculate the mean absorbance for each set of duplicate standards, controls and samples, and subtract the average zero standard optical density. Plot the standard curve on log-log graph paper or using Sigma plot software, with standard concentration on the x-axis and absorbance on the y-axis. Draw the best-fit straight line through the standard points.
    Typical Data: These standard curves are for demonstration only. A standard curve must be run with each assay. Assay Diluent Human IgG1 concentration (ng/mL) O D =4 50 n m 0.1 1 10 0 0.1 1 10 100
    Sensitivity: The minimum detectable dose of IgG1 is typically less than 0.05 ng/mL. C. LINEARITY Sample Type Serum Plasma 1:2 Average % of Expected 88 82 Range ( %) 77-97 72-94 1:4 Average % of Expected 86 81 Range ( %) 75-96 71-92 D. REPRODUCIBILITY Intra-Assay: CV<10 % Inter-Assay: CV<12 %

    Testpräzision
    Intra-Assay: CV< 10 % Inter-Assay: CV< 12 %
    Beschränkungen
    Nur für Forschungszwecke einsetzbar
  • Handhabung
    Avoid repeated freeze-thaw cycles.
    Lagerung
    -20 °C
    Informationen zur Lagerung
    The entire kit may be stored at -20°C for up to 1 year from the date of shipment. Avoid repeated freeze-thaw cycles. The kit may be stored at 4°C for up to 6 months. For extended storage, it is recommended to store at -80°C.
    Haltbarkeit
    6 months
  • Olivares, Castillejo, Varea, Sanz: "Double-blind, randomised, placebo-controlled intervention trial to evaluate the effects of Bifidobacterium longum CECT 7347 in children with newly diagnosed coeliac disease." in: The British journal of nutrition, Vol. 112, Issue 1, pp. 30-40, (2014) (PubMed).

    Zhang, Tang, Goryacheva, Niessner, Knopp: "Anodic-stripping voltammetric immunoassay for ultrasensitive detection of low-abundance proteins using quantum dot aggregated hollow microspheres." in: Chemistry (Weinheim an der Bergstrasse, Germany), Vol. 19, Issue 7, pp. 2496-503, (2013) (PubMed).

    Prakash, Andrabi, Kumar, Kabra, Lodha, Vajpayee, Luthra: "Binding antibody responses to the immunogenic regions of viral envelope in HIV-1-infected Indian children." in: Viral immunology, Vol. 24, Issue 6, pp. 463-9, (2012) (PubMed).

    Kim, Lo, Wear, Stojadinovic, Weina, Izadjoo: "Improvement of anti-Burkholderia mouse monoclonal antibody from various phage-displayed single-chain antibody libraries." in: Journal of immunological methods, Vol. 372, Issue 1-2, pp. 146-61, (2011) (PubMed).

    Liao, Tsai: "Clinical effectiveness of Tyrophagus putrescentiae allergy by local nasal immunotherapy using strips of Dermatophagoides pteronyssinus." in: The Journal of asthma : official journal of the Association for the Care of Asthma, Vol. 48, Issue 9, pp. 957-64, (2011) (PubMed).

    Barry, Soloviev: "Quantitative protein profiling using antibody arrays." in: Proteomics, Vol. 4, Issue 12, pp. 3717-26, (2004) (PubMed).

  • Target Alle IgG1 ELISA Kits anzeigen
    IgG1
    Abstract
    IgG1 Produkte
    Synonyme
    IgG1 ELISA Kit, Igh-4 ELISA Kit, VH7183 ELISA Kit, immunoglobulin heavy constant gamma 1 (G1m marker) ELISA Kit, Ighg1 ELISA Kit
    Substanzklasse
    Antibody
    Hintergrund
    The human immune system consists of two functional components classified as the innate system (the physical, biochemical and cellular barriers), and the adaptive immune system (including lymphocytes and immunoglobulins). Immunoglobulins are the key elements of the humoral immune response in vertebrate against parasitic invasion. The polypeptide chains of immunoglobulins composed of two identical heavy (H) chains and two identical light (L) chains linked together by inter-chain disulfide bonds. While the amino-terminal portions that exhibits highly variable amino-acid composition are involved in antigen binding, the C terminal constant parts are involved in complement binding, placental passage and binding to cell membranes. Based upon the variation of the constant region of the heavy chain, nine immunoglobulin heavy chain isotypes are found in humans: IgA (with subclasses IgA1 and IgA2), IgD, IgE, IgM, and IgG (with subclasses IgG1, IgG2, IgG3, and IgG4). IgG is the predominant immunoglobulin in the serum, which accounts for 75% of the total serum antibody of healthy individuals. IgG has a molecular weight of about 150 kDa. Four distinct subgroups of human IgG (IgG1, IgG2, IgG3, and IgG4) were first demonstrated in the 1960™s by using polyclonal antisera prepared in animals immunized with human myeloma proteins. The Human IgG1 ELISA (Enzyme-Linked Immunosorbent Assay) kit is an in vitro enzyme-linked immunosorbent assay for the quantitative measurement of human IgG1 in serum and plasma. This assay employs an antibody specific for human IgG1 coated on a 96-well plate. Standards and samples are pipetted into the wells and IgG1 present in a sample is bound to the wells by the immobilized antibody. The wells are washed and biotinylated anti-human IgG antibody is added. After washing away unbound biotinylated antibody, HRP-conjugated streptavidin is pipetted to the wells. The wells are again washed, a TMB substrate solution is added to the wells and color develops in proportion to the amount of IgG1 bound. The Stop Solution changes the color from blue to yellow, and the intensity of the color is measured at 450 nm. II. Reproducibility: Intra-Assay: CV<10% Inter-Assay: CV<12%.
    UniProt
    P01857
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