Astana Biomed Group, an authorized Abcam distributor in Central Asia

Human C4b (Complement C4-B) ELISA Kit (ab277717)

Key features and details

  • One-wash 90 minute protocol
  • Sensitivity: 34.83 pg/ml
  • Range: 140.63 pg/ml - 9000 pg/ml
  • Sample type: Cell culture supernatant, Cerebral Spinal Fluid, Cit plasma, EDTA Plasma, Hep Plasma, Milk, Saliva, Serum, Urine
  • Detection method: Colorimetric
  • Assay type: Sandwich (quantitative)
  • Reacts with: Human

Overview

  • Product name

    Human C4b (Complement C4-B) ELISA Kit

  • Detection method

    Colorimetric

  • Precision

    Intra-assay
    Sample n Mean SD CV%
    Serum 8 1.9%
    Inter-assay
    Sample n Mean SD CV%
    Serum 3 3.9%

  • Sample type

    Cell culture supernatant, Saliva, Milk, Urine, Serum, Hep Plasma, EDTA Plasma, Cit plasma, Cerebral Spinal Fluid

  • Assay type

    Sandwich (quantitative)

  • Sensitivity

    34.83 pg/ml

  • Range

    140.63 pg/ml - 9000 pg/ml

  • Recovery

    Sample specific recovery
    Sample type Average % Range
    Cell culture supernatant 107 94% - 118%
    Saliva 88 80% - 89%
    Milk 92 91% - 94%
    Urine 109 99% - 118%
    Serum 92 89% - 95%
    Hep Plasma 96 93% - 100%
    EDTA Plasma 93 92% - 96%
    Cit plasma 101 97% - 103%
    Cerebral Spinal Fluid 106 102% - 109%

  • Assay time

    1h 30m

  • Assay duration

    One step assay

  • Species reactivity

    Reacts with: Human

  • Product overview

    Human C4b ELISA kit (ab277717) is a single-wash 90 min sandwich ELISA designed for the quantitative measurement of Human C4b protein in human serum, plasma, milk, saliva, urine, cerebrospinal fluid, and cell culture supernatant. It uses our proprietary SimpleStep ELISA® technology. Quantitate Human C4b with 34.83 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

    C4b is a non-enzymatic component of the C3 and C5 convertases and essential for the propagation of the classical complement pathway. C4b covalently binds to immunoglobulins and immune complexes and enhances the solubilization of immune aggregates and the clearance of IC through CR1 on erythrocytes. C4b catalyzes the transacylation of the thioester carbonyl group to form ester bonds with carbohydrate antigens. Mouse and rat share 76 % and 79 % sequence homology with human C4b.

  • 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 C4b (Complement C4-B) Capture Antibody 1 x 600µl
    10X Human C4b (Complement C4-B) Detector Antibody 1 x 600µl
    10X Wash Buffer PT (ab206977) 1 x 20ml
    Antibody Diluent 5BI 1 x 6ml
    Human C4b (Complement C4-B) 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

  • Function

    C4 plays a central role in the activation of the classical pathway of the complement system. It is processed by activated C1 which removes from the alpha chain the C4a anaphylatoxin. The remaining alpha chain fragment C4b is the major activation product and is an essential subunit of the C3 convertase (C4b2a) and the C5 convertase (C3bC4b2a) enzymes of the classical complement pathway.
    Derived from proteolytic degradation of complement C4, C4a anaphylatoxin is a mediator of local inflammatory process. It induces the contraction of smooth muscle, increases vascular permeability and causes histamine release from mast cells and basophilic leukocytes.

  • Sequence similarities

    Contains 1 anaphylatoxin-like domain.
    Contains 1 NTR domain.

  • Post-translational
    modifications

    Prior to secretion, the single-chain precursor is enzymatically cleaved to yield the non-identical chains (alpha, beta and gamma). During activation, the alpha chain is cleaved by C1 into C4a and C4b, and C4b stays linked to the beta and gamma chains. Further degradation of C4b by C1 into the inactive fragments C4c and C4d blocks the generation of C3 convertase.

  • Cellular localization

    Secreted.
  • Information by UniProt

  • Alternative names

    • Basic complement C4
    • C2B5
    • C3 and PZP-like alpha-2-macroglobulin domain-containing protein 3
    • C4b
    • C4B1
    • C4B2
    • C4B3
    • CO4B_HUMAN
    • Complement C4 gamma chain
    • Complement component 4B
    • CPAMD3
    see all

  • Database links

    see all

Images

  • SimpleStep ELISA Protocol Diagram
    SimpleStep ELISA Protocol Diagram

    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 human C4b standard curve in Sample Diluent NS. The C4b standard curve was prepared as described in Section 10.
    Example of human C4b standard curve in Sample Diluent NS. The C4b standard curve was prepared as described in Section 10.

    Raw data values are shown in the table. Background-subtracted data values (mean +/- SD) are graphed.

  • Interpolated concentrations of native C4b in human serum and plasma samples.
    Interpolated concentrations of native C4b in human serum and plasma samples.

    The concentrations of C4b were measured in duplicates, interpolated from the C4b standard curves and corrected for sample dilution. Undiluted samples are as follows: serum 1:50,000, plasma (citrate) 1:25,000, plasma (EDTA) 1:50,000, and plasma (heparin) 1:50,000. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean C4b concentration was determined to be 351.14 µg/mL in serum, 52.76 µg/mL in plasma (citrate) and 307.91 µg/mL in plasma (EDTA), and 156.35 µg/mL in plasma (heparin).  

     

     

  • Interpolated concentrations
    Interpolated concentrations

    Undiluted samples are as follows: saliva 1:25, HepG2 supernatant 1:20, urine 1:50, cerebrospinal fluid 1:500, and milk 1:500. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean C4b concentration was determined to be 135.80 ng/mL in saliva, 103.10 ng/mL in HepG2 supernatant, 408.43 ng/mL in urine, 2319.81 ng/mL in cerebrospinal fluid, and 3468.94 ng/mL in milk.

  • Serum diluted 1:100,000 from ten individual healthy human female donors was measured in duplicate.
    Serum diluted 1:100,000 from ten individual healthy human female donors was measured in duplicate.

    Interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean C4b concentration was determined to be 299.35 µg/mL with a range of 95.86 – 529.04 µg/mL.

  • Recombinant Antibody Benefits
    Recombinant Antibody Benefits

    To learn more about the advantages of recombinant antibodies see here.

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