概述

  • 产品名称
    人Complement C3 ELISA试剂盒
    参阅全部 C3 试剂盒
  • 检测方法
    Colorimetric
  • 精确度
    批次内
    样品 n Mean SD CV%
    Overall 3.1%
    批次间
    样品 n Mean SD CV%
    Overall 8.1%
  • 样品类型
    Saliva, Milk, Serum, Plasma
  • 检测类型
    Competitive
  • 灵敏度
    = 0.2 µg/ml
  • 范围
    0.47 µg/ml - 30 µg/ml
  • 回收率

    99 %

  • 检测时间
    3h 00m
  • 实验步骤
    Multiple steps standard assay
  • 种属反应性
    与反应: Human
  • 产品概述

    Complement C3 Human in vitro competitive ELISA (Enzyme-Linked Immunosorbent Assay) kit is designed for the quantitative measurement of Complement C3 in human plasma and serum.


    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 protein 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 inversely proportional to the amount of Complement C3 captured in plate.

  • 经测试应用
    适用于: Competitive ELISAmore details
  • 平台
    Microplate

性能

  • 存放说明
    Store at -20°C. Please refer to protocols.
  • 组件 1 x 96 tests
    100X Streptavidin-Peroxidase Conjugate 1 x 80µl
    10X Diluent M Concentrate 1 x 30ml
    1X Biotinylated Human Complement C3 (Lyophilized) 1 vial
    20X Wash Buffer Concentrate 1 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
  • 研究领域
  • 功能
    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.
  • 组织特异性
    Plasma.
  • 疾病相关
    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.
  • 序列相似性
    Contains 1 anaphylatoxin-like domain.
    Contains 1 NTR domain.
  • 翻译后修饰
    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.
  • 细胞定位
    Secreted.
  • Information by UniProt
  • 别名
    • 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
    • CO3_HUMAN
    • Complement C3
    • Complement C3c alpha'' chain fragment 2
    • Complement component 3
    • Complement factor 3
    • CPAMD1
    • HEL S 62p
    see all
  • 数据库链接

应用

Our Abpromise guarantee covers the use of ab108822 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

应用 Ab评论 说明
Competitive ELISA Use at an assay dependent concentration.

图片

  • Compl. C3 measured in biological fluids showing quantity (micrograms) per mL of tested sample

  • Representative Standard Curve using ab108822.

实验方案

文献

This product has been referenced in:
  • McGuire JL  et al. The complement system, neuronal injury, and cognitive function in horizontally-acquired HIV-infected youth. J Neurovirol 22:823-830 (2016). ELISA ; Human . Read more (PubMed: 27273074) »
  • Natrajan MS  et al. Pioglitazone regulates myelin phagocytosis and multiple sclerosis monocytes. Ann Clin Transl Neurol 2:1071-84 (2015). ELISA ; Human . Read more (PubMed: 26734659) »
See all 3 Publications for this product

客户评价及客户问答

1-10 of 12 Abreviews or Q&A

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


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

Thank you for contacting us. The laboratory does not think the background will be a problem with not diluting the samples. However, they did indicate that ab108823 may be more sensitive for detecting other species.


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

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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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Answer

Thank you for contacting us.

I can confirm that our Complement C3 Human ELISA kit (ab108822) is tested to work with human serum and plasma and covered by our 6 month guarantee in case it does not work. Could you confirm if that is the kit you are using?

We would not expect a high background for such high dilutions like 1:20000 because the recommended dilution for serum samples is 1:800 into diluent. The recommended dilution is stated in the protocol booklet. To read the whole protocol booklet click https://www.abcam.com/ps/products/108/ab108822/documents/ab108822%20Complement%20C3%20Human%20ELISA%20Kit%20v2%20(website).pdf.

We take problems with our products very seriously, and investigate every product that we feel may not be performing correctly. I would like to investigate the problem further. I would appreciate if you could take some time to answer our questionnaire. Could you also please send me an example of your results to give me some more insight into the problem?

A. General Information
- Catalogue number:

- Lot number:

- Purchase order number/ order date or preferably the Abcam order number:

- How was the kit stored?


B. Description of the problem
- Are the signals you obtained with control or standard satisfactory? (Yes/No);


- Are the sample readings as expected or inconsistent?


- Are the blank readings as expected?

- Other (e.g. missing components)


C. What do you think the problem is caused by? E.g. enzyme is faulty, developer is faulty or other component that you think might be faulty.


Thank you for your cooperation. I look forward hearing from you again and hope I can help you after receiving more information.


Use our products? Submit an Abreview. Earn rewards!
https://www.abcam.com/abreviews

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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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Answer

Gracias por contactar con abcam.

Me gustaría informarle que tenemos 2 kits para la deteccion de humano C3 (tambien llamado ASP).

Las referencias sonab108822 (para Serum, Plasma) and ab108823 (para Cell culture supernatant, Saliva, Milk, Urine).

Los links a las hojas de datos y a los protocoloslos encontrara a continuación:

- ab108822:
Datasheet: https://www.abcam.com/ab108822
Protocol Booklet: https://www.abcam.com/ps/products/108/ab108822/documents/ab108822%20Complement%20C3%20Human%20ELISA%20Kit%20(Website).pdf

- ab108823:
Datasheet: https://www.abcam.com/ab108823
Protocol booklet: https://www.abcam.com/ps/products/108/ab108823/documents/ab108823%20Complement%20C3%20Human%20ELISA%20Kit%20(Web)%20v2.pdf

Informarle que los precios para los productos así como los gastos de envíoque aparecen ennuestra página web están totalmente actualizados.

Esperamos que esta información le haya sido de utilidad. Por favor no dude en ponerse en contacto con nosotros si necesita algo mas.

Reciba un cordial saludo.

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Question
Answer

Thank you for contacting us in regards to Complement C3 Human ELISA kit (ab108822).


I now have some further information to share with you concerning the specificity of the kit. The kit is able torecognize complement C3a and other isoforms of complement C3. The measurement made using this kit is to determine the total complement C3 present in the sample.


I hope this information has been of help. If you require any further information please do not hesitate to contact us again.

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Answer

Thank you for contacting us and pointing out the inconsistency in the protocol provided with the Complement C3 Human ELISA kit (ab108822).


I can confirm that the standard solutions should be made to ug/mL not ng/mL as state in the table of the protocol. This will be updated on the protocol sheet.


I hope this information has been of help. If you require any further information please do not hesitate to contact us again. Otherwise, I wish you all the best with your research.

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1-10 of 12 Abreviews or Q&A

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

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