Overview

  • Product name

    Human Complement C3 ELISA Kit
    See all C3 kits
  • Detection method

    Colorimetric
  • Precision

    Intra-assay
    Sample n Mean SD CV%
    Overall 3.5%
    Inter-assay
    Sample n Mean SD CV%
    Overall 10%
  • Sample type

    Cell culture supernatant, Saliva, Milk, Urine, Cell Lysate, Cerebral Spinal Fluid
  • Assay type

    Sandwich (quantitative)
  • Sensitivity

    = 0.3 ng/ml
  • Range

    0.625 ng/ml - 40 ng/ml
  • Recovery

    97 %

  • Assay time

    4h 00m
  • Assay duration

    Multiple steps standard assay
  • Species reactivity

    Reacts with: Human
  • Product overview

    Human Complement C3 ELISA kit is designed for the quantitative measurement of Complement C3 concentrations in Human urine, milk, saliva, cerebrospinal fluid and cell culture supernatants.


    A Complement C3 specific antibody has been precoated onto 96-well plates and blocked. Standards or test samples are added to the wells and subsequently a Complement C3 specific biotinylated detection antibody is added and then followed by washing with wash buffer. Streptavidin-Peroxidase Conjugate is added and unbound conjugates are washed away with wash buffer. TMB is then used to visualize Streptavidin-Peroxidase enzymatic reaction. TMB is catalyzed by Streptavidin-Peroxidase to produce a blue color product that changes into yellow after adding acidic stop solution. The density of yellow coloration is directly proportional to the amount of Complement C3 captured in plate.


    The entire kit may be stored at -20°C for long term storage before reconstitution - Avoid repeated freeze-thaw cycles.

  • Tested applications

    Suitable for: Sandwich ELISAmore details
  • Platform

    Microplate

Properties

  • Storage instructions

    Store at -20°C. Please refer to protocols.
  • Components 1 x 96 tests
    100X Biotinylated Human Complement C3 Antibody 1 x 60µl
    100X Streptavidin-Peroxidase Conjugate 1 x 80µl
    10X Diluent N Concentrate 1 x 30ml
    20X Wash Buffer Concentrate 2 x 30ml
    Chromogen Substrate 1 x 8ml
    Complement C3 Microplate (12 x 8 well strips) 1 unit
    Complement C3 Standard 1 vial
    Sealing Tapes 3 units
    Stop Solution 1 x 12ml
  • Research areas

  • Function

    C3 plays a central role in the activation of the complement system. Its processing by C3 convertase is the central reaction in both classical and alternative complement pathways. After activation C3b can bind covalently, via its reactive thioester, to cell surface carbohydrates or immune aggregates.
    Derived from proteolytic degradation of complement C3, C3a 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.
  • Tissue specificity

    Plasma.
  • Involvement in disease

    Defects in C3 are the cause of complement component 3 deficiency (C3D) [MIM:120700]. A rare defect of the complement classical pathway. Patients develop recurrent, severe, pyogenic infections because of ineffective opsonization of pathogens. Some patients may also develop autoimmune disorders, such as arthralgia and vasculitic rashes, lupus-like syndrome and membranoproliferative glomerulonephritis.
    Genetic variation in C3 is associated with susceptibility to age-related macular degeneration type 9 (ARMD9) [MIM:611378]. ARMD is a multifactorial eye disease and the most common cause of irreversible vision loss in the developed world. In most patients, the disease is manifest as ophthalmoscopically visible yellowish accumulations of protein and lipid that lie beneath the retinal pigment epithelium and within an elastin-containing structure known as Bruch membrane.
    Defects in C3 are a cause of susceptibility to hemolytic uremic syndrome atypical type 5 (AHUS5) [MIM:612925]. An atypical form of hemolytic uremic syndrome. It is a complex genetic disease characterized by microangiopathic hemolytic anemia, thrombocytopenia, renal failure and absence of episodes of enterocolitis and diarrhea. In contrast to typical hemolytic uremic syndrome, atypical forms have a poorer prognosis, with higher death rates and frequent progression to end-stage renal disease. Note=Susceptibility to the development of atypical hemolytic uremic syndrome can be conferred by mutations in various components of or regulatory factors in the complement cascade system. Other genes may play a role in modifying the phenotype.
  • Sequence similarities

    Contains 1 anaphylatoxin-like domain.
    Contains 1 NTR domain.
  • Post-translational
    modifications

    C3b is rapidly split in two positions by factor I and a cofactor to form iC3b (inactivated C3b) and C3f which is released. Then iC3b is slowly cleaved (possibly by factor I) to form C3c (beta chain + alpha' chain fragment 1 + alpha' chain fragment 2), C3dg and C3f. Other proteases produce other fragments such as C3d or C3g.
    Phosphorylation sites are present in the extracelllular medium.
  • Cellular localization

    Secreted.
  • Information by UniProt
  • Alternative names

    • Acylation stimulating protein cleavage product
    • AHUS5
    • ARMD9
    • ASP
    • C3
    • C3 and PZP like alpha 2 macroglobulin domain containing protein 1
    • C3 and PZP-like alpha-2-macroglobulin domain-containing protein 1
    • c3 complement
    • C3adesArg
    • CO3_HUMAN
    • Complement C3
    • Complement C3 alpha chain
    • Complement C3b alpha' chain
    • Complement C3c alpha' chain fragment 1
    • Complement C3c alpha' chain fragment 2
    • Complement C3c alpha'' chain fragment 2
    • Complement C3d fragment
    • Complement C3dg fragment
    • Complement C3f fragment
    • Complement C3g fragment
    • Complement component 3
    • Complement factor 3
    • CPAMD1
    • HEL S 62p
    • omplement C3 beta chain
    see all
  • Database links

Applications

Our Abpromise guarantee covers the use of ab108823 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

  • Complement C3 measured in various samples showing quantity (microgram) per mL of tested sample

  • Representative Standard Curve using ab108823

Protocols

References

This product has been referenced in:

  • Chang TT  et al. Plasma proteome plus site-specific N-glycoprofiling for hepatobiliary carcinomas. J Pathol Clin Res 5:199-212 (2019). Read more (PubMed: 31136099) »
  • Merchant ML  et al. Biomarker enhanced risk prediction for development of AKI after cardiac surgery. BMC Nephrol 19:102 (2018). Read more (PubMed: 29720115) »
See all 4 Publications for this product

Customer reviews and Q&As

1-10 of 13 Abreviews or Q&A

human C3 Elisa Kit

Excellent Excellent 5/5 (Ease of Use)
Abreviews
A good kit, works safely and reliably. We used the kit with cell supernatants. It's easy to handle compared to other Elisa kits and the instructions are easy to understand and follow.

Abcam user community

Verified customer

Submitted Aug 30 2019

good Elisa

Excellent Excellent 5/5 (Ease of Use)
Abreviews
The protocol booklet is well explained, easy to understand and easy to perform.
I used cell culture supernatants.
The kit is easy to use and quick compared to other Sandwich Elisa.
We obtain reliable and reproducible results with this Elisa.

Abcam user community

Verified customer

Submitted Feb 08 2018

Answer

The first assay you asked about, ab108823, is that kind of assay (sandwich), but the secondary (detection) antibody is biotinylated, not conjugated to an enzyme. The enzyme (HRP) is conjugated to streptavidin, which binds to the biotin on the antibody.

The ab108823 assay is very sensitive, and C3 concentrations in plasma are high compared to the C3 concentrations of the sample types it is recommended for: supernatants, saliva, milk and urine. The assay can be used with plasma samples but they will need to be diluted in order to be within the range of the standards. The average concentration of C3 in normal human plasma is about 1.5 mg/ml. The range of standards in ab108823 is 0.625 - 40 ng/ml. So, you will need to dilute your samples approximately 1/100,000 - 1/1000,000 to be within the range of the standards.

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Answer

A better choice for plasma samples is ab108822.

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Answer


Complement C3 is very abundant in plasma/serum samples. We have developed two different assays.

ab108823 is a sensitive sandwich ELISA assay. If you use this kit for plasma or serum sample, it requires very large dilution. Normal plasma C3 is average about 900 ug/ml. Our highest standard for ab108823 is 40 ng/ml. It requires a lot of dilution in order for sample to fall within the standard curve for kit ab108823. The more dilution, the less accurate it is for your assay and the higher the variation. That is why we develop the less sensitive assay ab108822 to address this issue.

ab108822 reduces the dilution factor significantly. The assay is more reproducible than ab108823 for plasma.

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Answer



Unfortunately, I can confirm that the antibodies used in these kits have been raised against recombinant protein and purified proteins. The exact epitope of the antibodies have not been investigated further. We are therefore not able to provide you with any information as to the site of recognition involved.

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Answer

I am sorry to hear that you have experienced so many problems with the Complement C3 Human ELISA kit (ab108823). I appreciate the time you took to provide further details.

After reviewing the provided information I suggest to verify the samples type that was used with the kit.

We offer two different Kits for Complement C3 Human. One can be used for cell culture supernatant, saliva, milk and urine. That is the ab108823 that was used in the experiments. The second kit is ab108822. This is made for the use with serum and plasma samples. The protocol and preparation of samples is different as well as the preparation of the standard curve to analyse the results.

However, I contacted the laboratory for you and can confirm that the ab108823 can be used with plasma and serum, but the samples need to be diluted.

It would require at least a 14000x dilution just to get a low range sample to within the upper range of the curve. And a 60000x or 120000x dilution to get it mid-curve.

I hope this information is helpful for you. I wish you all the best luck for future experiments. If you continue to have any difficulties please let me know.

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Question
Answer

Thank you for contacting us.

I have been in touch with the lab to inquire aboout use ofab108823 Complement C3 Human ELISA kit with plasma.This kit can be used for plasma, but you need to find out the appropriate dilution factor. We suggest to start with a few series dilution to find out best dilution factor. For example, 1:1000. 1:10000, 1:100000 to find the best dilution for plasma.

I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.

Use our products? Submit an Abreview. Earn rewards!
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Answer

Muchas gracias por tu respuesta.

Actualmente tenemos una promoción para España por la que se aplica el 25% de descuento a todos los productos que no son anticuerpos primarios. La información sobre la promoción está disponible en el siguiente link:

https://www.abcam.com/index.html?pageconfig=resource&rid=15194

Si necesitáis que os mandemos una proforma o cualquier información adicional, no dudéis en contactarnos.

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Answer

Siento no haber contestado antes, me temo que no tengo la información de todos los productos solicitados, sin embargo, puedo proporcionarte acerca de los siguientes kits:

El kit PLTP Activity Fluorometric Assay Kit (ab65386) no es apropiado para vuestro propósito puesto que reacciona con el ortólogo murino.

Para LIF Human ELISA Kit (ab100582) no se han llevado a cabo tests de reactividad cruzada, por lo que se desconoce si reconoce la proteína murina. El anticuerpo de captura es monoclonal y el de detección policlonal. Ambas proteínas comparten un 78% de homología, por lo que algo de reactividad cruzada es de esperar.

Respecto a Complement C3 Human ELISA kit y Fibronectin Human ELISA Kit:

ab108823: se ha observado una reactividad cruzada del 0.5% con ratón y del 10% con rata.

ab108822: ninguna reactividad cruzada observada.

ab108847: no se ha observado reactividad cruzada con el ortólogo murino

ab108848: reactividad del 5% con ratón y del 20% con rata.

Con Fibronectin Human ELISA kit (with color giving dyes) (ab119524) se ha encontrado cierta reactividad cruzada de acuerdo con el colaborador que nos lo suministra.

Para ab119611 y ab119612 (uPA Human ELISA kit y uPA Receptor Human ELISA Kit) no se ha encontrado reactividad con la proteína de ratón.

No tengo de momento información acerca de ab133037 pero si sigues interesado, en cuanto la tenga te contacto de nuevo.

Si necesitáis un presupuesto o más información acerca de cualquiera de las opciones ofrecidas, por favor, no dudéis en contactarnos de nuevo y estaré encantada de poder ayudar.

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1-10 of 13 Abreviews or Q&A

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